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WO2021226442A2 - Therapeutic uses of c3-binding agents - Google Patents

Therapeutic uses of c3-binding agents Download PDF

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Publication number
WO2021226442A2
WO2021226442A2 PCT/US2021/031268 US2021031268W WO2021226442A2 WO 2021226442 A2 WO2021226442 A2 WO 2021226442A2 US 2021031268 W US2021031268 W US 2021031268W WO 2021226442 A2 WO2021226442 A2 WO 2021226442A2
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WO
WIPO (PCT)
Prior art keywords
seq
amino acid
acid sequence
set forth
sequence set
Prior art date
Application number
PCT/US2021/031268
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French (fr)
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WO2021226442A3 (en
Inventor
Zhonghao LIU
Hui Tian
Original Assignee
Ngm Biopharmaceuticals, Inc.
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Publication of WO2021226442A2 publication Critical patent/WO2021226442A2/en
Publication of WO2021226442A3 publication Critical patent/WO2021226442A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present disclosure generally relates to methods of treating, preventing, or inhibiting the development of respiratory illness and diseases.
  • the disclosure relates to treating, preventing, or inhibiting the development of thrombosis and resulting sequelae such as stroke.
  • the methods comprise administering agents that bind complement component C3 (C3), particularly antibodies that bind human C3.
  • C3 complement component C3
  • the complement system functions as an immune surveillance system and is part of the innate immune response that rapidly responds to bacterial and/or viral infections. Activation of the complement system promotes inflammation, both acute and chronic, and often results in elimination of pathogens.
  • the complement system includes more than 30 cell-associated and circulating proteins (e.g ., Cl, Clq, Clr, Cls, C2, C3, C3a, C3b, C4, C5, Factor B, Factor D, Factor H, Factor I).
  • CP classical pathway
  • MBL lectin pathway
  • AP alternative pathway
  • the three complement pathways are initiated by different factors, each resulting in the cleavage of complement component C3.
  • the classical complement pathway is activated when the Cl complex binds specific antigen-antibody complexes, often immunoglobulin M (IgM), IgG3, or IgGl. This induces a conformational change in the Cl complex, allowing it to cleave C4 and C2 to generate the C4bC2b complex.
  • C4bC2b acts as the C3 convertase of the classical pathway.
  • the lectin complement pathway is activated when mannose-binding lectin (MBL) binds mannose-containing polysaccharides on microorganisms, initiating the cleavage of C4 and C2 by the MBL- MBL-associated serine protease complex.
  • MBL mannose-binding lectin
  • C4bC2b complex forms the C3 convertase of the lectin pathway.
  • the oldest evolutionary signaling pathway of the three, the alternative complement pathway acts both independently of, and as an amplification loop for, the classical and lectin pathways.
  • the alternative complement pathway undergoes low-level self-activation through the slow, spontaneous hydrolysis of C3.
  • C3(H20) binds complement Factor B, which is subsequently cleaved by complement Factor D into C3(H20)Bb, forming the C3 convertase of the alternative complement pathway.
  • C3b participates in the formation of the C5 convertase which, in turn, cleaves C5 to C5a and C5b.
  • C3a and C5a termed anaphylatoxins, are pleiotropic inflammatory mediators.
  • MAC membrane attack complex
  • the coagulation system functions to mediate blood clotting. Similar to the immune system, it is a highly complex system made up of a large number of blood cells and soluble factors/proteins. Normal coagulation system homeostasis represents a balance between (i) the pro-coagulant processes that are responsible for clot formation, (ii) mechanisms that inhibit the clotting process beyond an injury site, and (iii) fibrolysis, which involves a distinct enzymatic cascade that lead to removal of clots. Imbalance of the coagulation system and/or fibrolysis may occur during critical illness, in post-operative situations, or in some diseases. Dysfunction of the coagulation system is a major contributor to thrombosis - the pathological formation of thrombi in blood vessels.
  • a pro-inflammatory response is often observed as part of the normal immune response to a viral infection.
  • the pro-inflammatory response includes the production of cytokines and chemokines, which in turn recruit additional immune cells including leukocytes, macrophages, monocytes, and dendritic cells.
  • additional immune cells including leukocytes, macrophages, monocytes, and dendritic cells.
  • the subsequent effector functions of these effector cells and soluble cellular factors contribute to the first line of defense against the virus.
  • these events contribute to the reduction of viral replication and to the limitation of viral spread.
  • the viral infection induces an overly aggressive pro-inflammatory response, which in combination with an insufficient balancing of an anti-inflammatory response, can lead to a series of events called a “cytokine storm”.
  • Symptoms of a cytokine storm include, but are not limited to, fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rashes, fast breathing, rapid heartbeat, low blood pressure, seizures, headache, confusion, delirium, hallucinations, tremor, and loss of coordination.
  • a cytokine storm is often associated with a surge of activated immune cells into the lungs and over-production of cytokines/chemokines. The resulting lung inflammation and fluid buildup can lead to respiratory distress, lung damage, a variety of acute respiratory diseases, and even death.
  • Influenza viruses include, but are not limited to, H5N1 and H1N1 2009.
  • Coronaviruses include, but are not limited to, SARS-CoV-1 (severe acute respiratory syndrome coronavirus 1), SARS-CoV-2, and MERS-CoV (Middle East respiratory syndrome coronavirus). Infection with a coronavirus may lead to severe disease and the syndromes are generally referred to as SARS (SARS-CoV-1), COVID-19 (SARS-CoV- 2), and MERS (MERS-CoV).
  • the present disclosure provides methods of treating, preventing, or inhibiting the development of respiratory illnesses and diseases.
  • the methods relate to treating, preventing, or inhibiting the development of severe respiratory illnesses and diseases.
  • the disclosure provides methods of treating, preventing, or inhibiting the development of thrombosis and resulting sequelae such as stroke.
  • a respiratory illness is associated with elevated levels of pro-inflammatory cytokines and/or chemokines.
  • a severe respiratory illness is associated with elevated levels of pro- inflammatory cytokines and/or chemokines.
  • thrombosis is associated with elevated levels of pro-inflammatory cytokines and/or chemokines.
  • a respiratory illness is associated, either directly or indirectly, with dysregulation of the complement pathway.
  • a severe respiratory illness is associated, either directly or indirectly, with dysregulation of the complement pathway.
  • thrombosis is associated, either directly or indirectly, with dysregulation of the complement pathway.
  • the dysregulation of the complement pathway is associated with elevated levels of pro- inflammatory cytokines.
  • the respiratory illness is associated with a suspected or confirmed viral infection.
  • the severe respiratory illness is associated with a suspected or confirmed viral infection.
  • the thrombosis or sequelae thereof is associated with a suspected or confirmed viral infection.
  • the viral infection is caused by an influenza virus, a coronavirus, or a respiratory syncytial virus.
  • the suspected or confirmed viral infection is a coronavirus infection.
  • the suspected or confirmed viral infection is a SARS-CoV-2 infection.
  • a respiratory illness is associated with a suspected or confirmed coronavirus infection.
  • a severe respiratory illness is associated with a suspected or confirmed coronavirus infection.
  • a respiratory illness is associated with a suspected or confirmed SARS-CoV-2 infection.
  • a severe respiratory illness is associated with a suspected or confirmed SARS-CoV-2 infection.
  • thrombosis or sequelae thereof is associated with a suspected or confirmed coronavirus infection.
  • thrombosis or sequelae thereof is associated with a suspected or confirmed SARS-CoV-2 infection.
  • a method of treating a respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of an agent that specifically binds complement component C3 (referred to herein as a “C3 -binding agent”).
  • a method of treating a severe respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of an agent that specifically binds complement component C3.
  • a method of preventing or inhibiting the development of a respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of a severe respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • the severe respiratory illness is selected from the group consisting of: pneumonia, acute respiratory disease (ARD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), atypical ARDS, respiratory distress syndrome (RDS), severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus 2019 (COVID-19).
  • a method of treating ARDS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of ARDS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • the subject has a suspected or confirmed viral infection.
  • a method of inhibiting complement pathway activation in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of inhibiting complement pathway activation in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent, wherein the subject has a suspected or confirmed viral infection.
  • a method of treating thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of treating thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent, wherein the subject has a suspected or confirmed viral infection. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent, wherein the subject has a suspected or confirmed viral infection.
  • the thrombosis is a pulmonary embolism (PE), large blood vessel blood clot, a deep vein thrombosis (DVT), or microvascular thrombosis.
  • the thrombosis has the potential to lead to a stroke.
  • the subject has symptoms of a stroke or has been diagnosed with a stroke.
  • the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus.
  • the viral infection is caused by a coronavirus.
  • the coronavirus is selected from the group consisting of: SARS-CoV-2, SARS-CoV-1, or MERS-CoV.
  • the coronavirus is SARS-CoV-2.
  • the viral infection is caused by SARS-CoV-2.
  • the subject has been diagnosed with the syndrome/disease that is caused by SARS-CoV-2 - COVID-19.
  • a method of treating a coronavirus infection in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of treating a coronavirus infection or the resulting complications and/or sequelae of a coronavirus infection in a human subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent.
  • the coronavirus is selected from the group consisting of: SARS- CoV-2, SARS-CoV-1, or MERS-CoV.
  • the coronavirus is SARS-CoV-2.
  • an infection with SARS-CoV-2 results in COVID-19.
  • a virus infection is associated with a dysregulated pro-inflammatory cytokine response or cytokine storm.
  • a coronavirus infection is associated with a dysregulated pro- inflammatory cytokine response or cytokine storm.
  • a method of treating SARS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of SARS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • the subject has a suspected or confirmed SARS-CoV-1 infection.
  • a method of treating MERS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of MERS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • the subject has a suspected or confirmed MERS-CoV infection.
  • a method of treating COVID-19 in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • a method of preventing or inhibiting the development of COVID-19 in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent.
  • the subject has a suspected or confirmed SARS-CoV-2 infection.
  • the subject has lung damage, respiratory failure, kidney damage, kidney failure, liver damage, heart damage, vascular damage, thrombosis, stroke, central nervous system injury, and/or multiple organ failure.
  • the subject has one or more symptoms selected from the group consisting of: hypoxemia, cough, wheezing, dyspnea, hyperpnea, pulmonary /lung inflammation, shortness of breath, labored breathing, rapid breathing, accumulation of alveolar fluid, pulmonary edema, vascular leakage, lymphocyte infiltration in the lung, lymphopenia, fever, chills, shaking chills, increased heart rate, chest pain, low blood pressure, headache, confusion, seizures, extreme tiredness, sepsis, bluish coloring of nails or lips, toe rashes/redness, toe swelling, loss of sense of smell, loss of sense of taste, and diarrhea.
  • the subject has mild, moderate or severe hypoxemia as determined by Partial Pressure of arterial oxygen/Fraction of inspired oxygen (PaC 2 /FiC 2 ) or positive end-expiratory pressure (PEEP). In some embodiments of the methods described herein, the subject has severe hypoxemia with a PaCh/FiCh of less than 100.
  • a method comprises administering the C3 -binding agent as part of a combination therapy.
  • the combination therapy includes the use of a medical device.
  • the medical device is a ventilator or an ECMO machine.
  • the combination therapy includes the use of at least one additional therapeutic agent.
  • the additional therapeutic agent is one or more of: dexamethasone, remdesivir, baricitinib in combination with remdesivir, favipiravir, merimepodib, an anticoagulation drug selected from low-dose heparin or enoxaparin, bamlanivimab, a combination of bamlanivimab and etesevimab, a combination of casirivimab and imdevimab, convalescent plasma, an mRNA SARS-CoV-2 vaccine (e.g., those produced by Moderna and Pfizer), an attenuated SARS-CoV-2 virus vaccine, a dead SARS-CoV-2 virus vaccine, a viral vaccine against SARS-CoV-2 (e.g., an adenoviral vaccine such as those produced by Johnson & Johnson and Astrazeneca), an antibody or fragment thereof or small molecule that blocks interaction between hACE2 and the spike protein of SARS-CoV-2, protea
  • a method comprises administering to the subject a C3-binding agent described herein and at least one additional therapeutic agent.
  • the subject is human.
  • the present disclosure provides one or more agents that bind C3, referred to herein as “a C3-binding agent” or “C3-binding agents.”
  • the C3-binding agents bind human C3 (e.g., SEQ ID NO:l or SEQ ID NO:2).
  • the C3-binding agents bind cynomolgus monkey (“cyno”) C3 (e.g., SEQ ID NO:7 or SEQ ID NO:8).
  • the C3- binding agents bind human C3 and cyno C3.
  • the C3-binding agents comprise an antibody.
  • the C3-binding agents comprise an antibody that binds human C3. In some embodiments, the C3-binding agents comprise an antibody that binds cyno C3. In some embodiments, the C3-binding agents comprise an antibody that binds human C3 and cyno C3. In some embodiments, the C3-binding agents comprise an antibody that binds human C3 and does not bind human C3b. In some embodiments, the C3-binding agents do not bind human C3b at a detectable level.
  • the C3 -binding agents bind human C3 and have at least one or more (e.g, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) of the following properties: (a) binds to cyno C3, (b) is an antagonist of C3, (c) inhibits C3 activity, (d) inhibits C3 cleavage, (e) inhibits C3 cleavage and C3a release, (f) inhibits complement activation, (g) inhibits activation of the alternative complement pathway, (h) inhibits activation of the classical complement pathway, (i) inhibits activation of the alternative complement pathway and inhibits activation of the classical complement pathway, (j) does not detectably bind to Factor Bb, (k) does not detectably bind to C3d, and (1) does not detectably bind to C3a.
  • the C3 -binding agents inhibit activation of the classical complement pathway. In some embodiments, the C3-binding agents inhibit activation of the alternative complement pathway. In some embodiments, the C3 -binding agents inhibit activation of the alternative complement pathway and classical complement pathway.
  • a C3-binding agent comprises: (1) a heavy chain variable region (VH) comprising VH complementarity determining region (CDR)1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:31, and a light chain variable region (VL) comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:32; (2) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 33, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (3) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:34, and a VL comprising VL CDR1, VL CDR2, and
  • a C3 -binding agent (e.g., antibody) comprises (a) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 16, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:33;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:34;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77;
  • the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77;
  • the heavy chain variable region
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:34;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77;
  • the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:
  • the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:31; (b) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33; (c) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34; (d) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:222; (e) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:223; (f) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:224; (g) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:77; (h) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:95; (i) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 113; (j) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 131; (k) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:
  • a C3-binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3- binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprises a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO: 52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO
  • a C3-binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO:52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3- binding agent comprises a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNTGTTYNQQFTG (SEQ ID NO:48), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3 -binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHEGGTTYNQQFTG (SEQ ID NO:52), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the agent is an antibody.
  • the C3-binding agent is an anti-C3 antibody.
  • the present disclosure also provides an C3-binding agent that comprises a heavy chain variable region comprising the three VH CDRs according to any CDR definition provided in Table 1 A (for 38G10), Table IB (for Hz38G10 and Hz38G10(G56A)), and Table 1C (for Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q)) and a light chain variable region comprising the three VL CDRs according to any CDR definition provided in Table 1A (for 38G10), Table IB (for Hz38G10 and Hz38G10(G56A)), and Table 1C (for Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q)).
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to any one of SEQ ID NO: 31, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, and SEQ ID NO:224; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:32 or SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:31 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:32.
  • a C3 -binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:33 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:34 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:222 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:223 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:224 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region comprising SEQ ID NO:31 and a light chain variable region comprising SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:33 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:34 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:222 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:223 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:224 and a light chain variable region comprising SEQ ID NO:35.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from: (a) the antibody designated 38G10 that comprises a heavy chain variable region comprising SEQ ID NO:31 and a light chain variable region comprising SEQ ID NO:32; (b) the antibody designated Hz38G10 that comprises a heavy chain variable region comprising SEQ ID NO:33 and a light chain variable region comprising SEQ ID NO:35; (c) the antibody designated Hz38G10(G56A) that comprises a heavy chain variable region comprising SEQ ID NO:34 and a light chain variable region comprising SEQ ID NO:35; (d) the antibody designated Hz38G10 (G56T) that comprises a heavy chain variable region comprising SEQ ID NO:222 and a light chain variable region comprising SEQ ID NO:35; (e) the antibody designated Hz38G10 (N55E) that comprises a heavy chain variable region comprising
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41.
  • a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 3D8 that comprises a heavy chain variable region comprising SEQ ID NO:77 and a light chain variable region comprising SEQ ID NO:78.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:77 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:78.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 2.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:77; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:78.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:77; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:78.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:77; and a light chain variable region having the sequence of SEQ ID NO:78.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 3G8 that comprises a heavy chain variable region comprising SEQ ID NO:95 and a light chain variable region comprising SEQ ID NO:96.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 95 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:96.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 3.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:95; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:96.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:95; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:96.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:95; and a light chain variable region having the sequence of SEQ ID NO:96.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 1502 that comprises a heavy chain variable region comprising SEQ ID NO: 113 and a light chain variable region comprising SEQ ID NO: 114.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 113 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:l 14.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 4.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 113; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:l 14.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 113; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 114.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 113; and a light chain variable region having the sequence of SEQ ID NO: 114.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 27A8 that comprises a heavy chain variable region comprising SEQ ID NO: 131 and a light chain variable region comprising SEQ ID NO: 132.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 131 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO: 132.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 5.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 131; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 132.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 131; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 132.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 131; and a light chain variable region having the sequence of SEQ ID NO: 132.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 28C3 that comprises a heavy chain variable region comprising SEQ ID NO: 149 and a light chain variable region comprising SEQ ID NO: 150.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 149 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:150.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 6.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 149; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 150.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 149; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 150.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 149; and a light chain variable region having the sequence of SEQ ID NO: 150.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 38F5 that comprises a heavy chain variable region comprising SEQ ID NO: 167 and a light chain variable region comprising SEQ ID NO: 168.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 167 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO: 168.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 7.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 167; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 168.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 167; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 168.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 167; and a light chain variable region having the sequence of SEQ ID NO: 168.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 62B11 that comprises a heavy chain variable region comprising SEQ ID NO: 185 and a light chain variable region comprising SEQ ID NO: 186.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VEl CDR2, and VH CDR3 of SEQ ID NO: 185 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:186.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 8.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 185; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 186.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 185; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 186.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 185; and a light chain variable region having the sequence of SEQ ID NO: 186.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 62F2 that comprises a heavy chain variable region comprising SEQ ID NO:203 and a light chain variable region comprising SEQ ID NO:204.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:203 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:204.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 9.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:203; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:204.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:203; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:204.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:203; and a light chain variable region having the sequence of SEQ ID NO:204.
  • a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 63 A3 that comprises a heavy chain variable region comprising SEQ ID NO:220 and a light chain variable region comprising SEQ ID NO:221.
  • the C3-binding agent is an anti-C3 antibody.
  • a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:220 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:221.
  • a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 10.
  • a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:220; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:221.
  • a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:220; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:221.
  • a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:220; and a light chain variable region having the sequence of SEQ ID NO:221.
  • a C3 -binding agent is an antibody.
  • the antibody is a monoclonal antibody.
  • the antibody is a humanized antibody.
  • the antibody is a human antibody.
  • the antibody is a chimeric antibody.
  • the antibody is a whole or intact antibody.
  • the antibody is an IgG antibody.
  • the antibody is an IgGl antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody.
  • the antibody is a human IgGl antibody, a human IgG2 antibody, a human IgG3 antibody, or a human IgG4 antibody.
  • the antibody comprises a human kappa chain or a human lambda light chain constant region.
  • the antibody is a bispecific antibody or a multispecific antibody.
  • the antibody is an antibody fragment.
  • the antibody or antibody fragment is a Fab, Fab’, F(ab’)2, Fv, scFv, (scFv)2, single chain antibody, dual variable region antibody, single variable region antibody, linear antibody, nanobody, or a V region antibody.
  • a C3-binding agent e.g, an antibody
  • the scFv comprises a heavy chain variable region comprising SEQ ID NO:225 or SEQ ID NO:226.
  • the scFv comprises a light chain variable region comprising SEQ ID NO:227.
  • the scFv comprises a heavy chain variable region comprising SEQ ID NO:225 and a light chain variable region comprising SEQ ID NO:227.
  • the scFv comprises a heavy chain variable region comprising SEQ ID NO:226 and a light chain variable region comprising SEQ ID NO:227.
  • the scFv comprises SEQ ID NO:228. In some embodiments, the scFv comprises SEQ ID NO:229. In some embodiments, the scFv comprises SEQ ID NO:230. In some embodiments, the scFv comprises SEQ ID NO:231.
  • the anti-C3 antibody is an antagonistic antibody. In some embodiments, the anti-C3 antibody inhibits C3 activity. In some embodiments, the anti-C3 antibody inhibits activation of the complement system. In some embodiments, the anti-C3 antibody inhibits activation of the classical complement pathway. In some embodiments, the anti-C3 antibody inhibits activation of the alternative complement pathway. In some embodiments, the anti-C3 antibody inhibits activation of the classical complement pathway and the alternative complement pathway. [0059] In another aspect, the disclosure provides compositions comprising a C3- binding agent described herein. In some embodiments, the disclosure provides compositions comprising an anti-C3 antibody described herein.
  • the disclosure provides pharmaceutical compositions comprising a C3-binding agent described herein and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides pharmaceutical compositions comprising an anti-C3 antibody described herein and a pharmaceutically acceptable carrier.
  • the C3-binding agent is isolated. In some embodiments, the C3-binding agent is substantially pure.
  • the disclosure provides polynucleotides comprising a polynucleotide or polynucleotides that encode a C3 -binding agent described herein.
  • the polynucleotide or polynucleotides are isolated.
  • a vector or vectors comprise the polynucleotide or polynucleotides that encode a C3-binding agent described herein.
  • an isolated cell comprises the polynucleotide or polynucleotides that encode a C3-binding agent described herein.
  • an isolated cell comprises the vector or vectors comprising a polynucleotide or polynucleotides that encode a C3-binding agent described herein.
  • the disclosure provides a cell comprising a C3-binding agent described herein.
  • the disclosure provides a cell producing a C3 -binding agent described herein.
  • a cell produces an anti-C3 antibody described herein.
  • a cell is a monoclonal cell line.
  • a cell is a hybridoma.
  • FIG. 1 Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody 38G10, Ab2, and Ab3 were tested for their ability to inhibit complement activation.
  • Figure 3 Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody Hz38G10 and compstatin derivative (COMP) were tested for their ability to inhibit complement activation.
  • COMP compstatin derivative
  • Figure 4 Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody Hz38G10 and a pegylated version of the compstatin derivative (COMP -PEG) were tested for their ability to inhibit complement activation.
  • COMP -PEG compstatin derivative
  • Figure 5 Binding Site Analysis. Hz38G10 and a compstatin derivative were tested in a competitive binding assay to assess whether the molecules bind to the same or different epitopes.
  • the present disclosure provides methods of treating respiratory illness, methods of preventing or inhibiting the development of respiratory illness, methods of inhibiting complement pathway activation, methods of treating thrombosis, methods of preventing or inhibiting the development of thrombosis, and methods of treating a coronavirus infection or the resulting complications and/or sequelae of a coronavirus infection.
  • the methods provided herein comprise administering to a subject agents that specifically bind human complement component C3 (i.e., a C3-binding agent).
  • the C3 -binding agents include, but are not limited to, polypeptides, antibodies and antigen-binding fragments thereof, scaffold proteins, and heterodimeric molecules.
  • C3-binding agents include, but are not limited to, antagonists of C3 activity, inhibitors of C3 activity, and/or agents that modulate C3 activity.
  • Related polypeptides, polynucleotides, vectors, compositions comprising the agents, cells comprising the related polynucleotides or vectors, and methods of making the agents are also provided and/or used in the methods described herein.
  • binding agent refers to a molecule which binds a specific antigen or target (e.g ., complement component C3).
  • a binding agent may comprise a protein, peptide, nucleic acid, carbohydrate, lipid, or small molecular weight compound.
  • a binding agent comprises an antibody.
  • a binding agent is an antibody.
  • a binding agent comprises an alternative protein scaffold or artificial scaffold (e.g., a nonimmunoglobulin backbone).
  • a binding agent is a fusion protein comprising an antigen-binding site.
  • a binding agent is a bispecific or multispecific molecule comprising at least one antigen-binding site.
  • antibody refers to an immunoglobulin molecule, or antigen-binding fragment thereof, that recognizes and binds a target through at least one antigen-binding site.
  • Antibody is used herein in the broadest sense and encompasses various antibody structures, including but not limited to, polyclonal antibodies, recombinant antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, bispecific antibodies, multispecific antibodies, diabodies, tribodies, tetrabodies, single chain Fv (scFv) antibodies, and antibody fragments as long as they exhibit the desired antigen-binding activity.
  • intact antibody or “full-length antibody” refers to an antibody having a structure substantially similar to a native antibody structure. This includes, for example, an antibody comprising two light chains each comprising a variable region and a light chain constant region (CL) and two heavy chains each comprising a variable region and at least a hinge region and heavy chain constant regions CHI, CH2, and CH3.
  • CL light chain constant region
  • antibody fragment refers to a molecule other than an intact antibody that comprises a portion of an antibody and generally at least one antigen-binding site.
  • antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, single chain antibody molecules (e.g, scFv), disulfide-linked scFv (dsscFv), nanobodies, diabodies, tribodies, tetrabodies, minibodies, dual variable domain antibodies (DVD), single variable domain antibodies (e.g, camelid antibodies), and bispecific or multispecific molecules formed from antibody fragments.
  • the term “monoclonal antibody” as used herein refers to a substantially homogenous antibody population involved in the highly specific recognition and binding of a single antigenic determinant or epitope.
  • the term “monoclonal antibody” encompasses intact and full-length monoclonal antibodies as well as antibody fragments (e.g, Fab, Fab', F(ab')2, Fv), single chain antibodies (e.g, scFv), fusion proteins comprising an antibody fragment, and any other modified immunoglobulin molecule comprising at least one antigen-binding site.
  • “monoclonal antibody” refers to such antibodies made by any number of techniques, including but not limited to, hybridoma production, phage library display, recombinant expression, and transgenic animals.
  • chimeric antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a first source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
  • humanized antibody refers to an antibody that comprises a human heavy chain variable region and a light chain variable region wherein the native CDR residues are replaced by residues from corresponding CDRs from a nonhuman antibody (e.g, mouse, rat, rabbit, or nonhuman primate), wherein the nonhuman antibody has the desired specificity, affinity, and/or activity.
  • a nonhuman antibody e.g, mouse, rat, rabbit, or nonhuman primate
  • one or more framework region residues of the human heavy chain or light chain variable regions are replaced by corresponding residues from nonhuman antibody.
  • humanized antibodies can comprise residues that are not found in the human antibody or in the nonhuman antibody. In some embodiments, these modifications are made to further refine and/or optimize antibody characteristics.
  • the humanized antibody comprises at least a portion of an immunoglobulin constant region (e.g ., hinge region, CHI, CH2, CH3, and/or Fc), typically that of a human immunoglobulin.
  • human antibody refers to an antibody that possesses an amino acid sequence that corresponds to an antibody produced by a human and/or an antibody that has been made using any of the techniques that are known to those of skill in the art for making human antibodies. These techniques include, but not limited to, phage display libraries, yeast display libraries, transgenic animals, recombinant protein production, and B-cell hybridoma technology.
  • epitopes and “antigenic determinant” are used interchangeably herein and refer to that portion of an antigen or target capable of being recognized and bound by a particular antibody.
  • epitopes can be formed both from contiguous amino acids and noncontiguous amino acids juxtaposed by tertiary folding of the protein.
  • Epitopes formed from contiguous amino acids also referred to as linear epitopes
  • epitopes formed by tertiary folding also referred to as conformational epitopes
  • An epitope typically includes at least 3, and more usually, at least 5, 6, 7, or 8-10 amino acids in a unique spatial conformation.
  • Epitopes can be predicted using any one of a large number of software bioinformatic tools available on the internet.
  • X-ray crystallography may be used to characterize an epitope on a target protein by analyzing the amino acid residue interactions of an antigen/antibody complex.
  • the term “specifically binds” as used herein refers to an agent (e.g., an antibody) that interacts more frequently, more rapidly, with greater duration, with greater affinity, or with some combination of the above to a particular antigen, epitope, protein, or target molecule than with alternative substances.
  • a binding agent that specifically binds an antigen can be identified, for example, by immunoassays, ELISAs, SPR ( e.g ., Biacore), or other techniques known to those of skill in the art.
  • an agent that specifically binds an antigen e.g., human C3 can bind related antigens (e.g, cyno C3).
  • a binding agent that specifically binds an antigen can bind the target antigen at a higher affinity than its affinity for a different antigen.
  • the different antigen can be a related antigen.
  • a binding agent that specifically binds an antigen can bind the target antigen with an affinity that is at least 20 times greater, at least 30 times greater, at least 40 times greater, at least 50 times greater, at least 60 times greater, at least 70 times greater, at least 80 times greater, at least 90 times greater, or at least 100 times greater, than its affinity for a different antigen.
  • a binding agent that specifically binds a particular antigen binds a different antigen at such a low affinity that binding cannot be detected using an assay described herein or otherwise known in the art.
  • affinity is measured using SPR technology in a Biacore system as described herein or as known to those of skill in the art.
  • polypeptide and “peptide” and “protein” are used interchangeably herein and refer to polymers of amino acids of any length.
  • the polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by nonamino acids.
  • the terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification.
  • polypeptides containing one or more analogs of an amino acid including but not limited to, unnatural amino acids, as well as other modifications known in the art. It is understood that, because the polypeptides of this disclosure may be based upon antibodies, the term “polypeptide” encompasses polypeptides as a single chain and polypeptides of two or more associated chains.
  • polynucleotide and nucleic acid and nucleic acid molecule are used interchangeably herein and refer to polymers of nucleotides of any length, and include DNA and RNA.
  • the nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase.
  • nucleic acids or polypeptides refer to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned (introducing gaps, if necessary) for maximum correspondence, not considering any conservative amino acid substitutions as part of the sequence identity.
  • the percent identity may be measured using sequence comparison software or algorithms or by visual inspection.
  • Various algorithms and software that may be used to obtain alignments of amino acid or nucleotide sequences are well- known in the art. These include, but are not limited to, BLAST, ALIGN, Megalign, BestFit, GCG Wisconsin Package, and variants thereof.
  • two nucleic acids or polypeptides of the disclosure are substantially identical, meaning they have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, and in some embodiments at least 95%, 96%, 97%, 98%, 99% nucleotide or amino acid residue identity, when compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection.
  • identity exists over a region of the sequences that is at least 10, at least 20, at least 20- 40, at least 40-60 nucleotides or amino acid residues, at least 60-80 nucleotides or amino acid residues in length or any integral value there between.
  • identity exists over a longer region than 60-80 nucleotides or amino acid residues, such as at least 80-100 nucleotides or amino acid residues, and in some embodiments the sequences are substantially identical over the full length of the sequences being compared, for example, (i) the coding region of a nucleotide sequence or (ii) an amino acid sequence.
  • amino acid substitution refers to a substitution in which one amino acid residue is replaced with another amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains have been generally defined in the art, including basic side chains (e.g ., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g, threonine, valine, isoleucine) and aromatic side chains (e.g, tyrosine, phenylalanine, tryptophan, hist
  • basic side chains e.g .,
  • substitution of a valine for a leucine is considered to be a conservative substitution.
  • conservative substitutions in the sequences of polypeptides and/or antibodies do not abrogate the binding of the polypeptide or antibody to the target binding site.
  • Methods of identifying nucleotide and amino acid conservative substitutions that do not eliminate binding are well-known in the art.
  • vector means a construct, which is capable of delivering, and usually expressing, one or more gene(s) or sequence(s) of interest in a host cell.
  • vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid, or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, and DNA or RNA expression vectors encapsulated in liposomes.
  • isolated refers to a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition which is in a form not found in nature.
  • An “isolated” antibody is substantially free of material from the cellular source from which it is derived.
  • isolated polypeptides, soluble proteins, antibodies, polynucleotides, vectors, cells, or compositions are those which have been purified to a degree that they are no longer in a form in which they are found in nature.
  • a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition which is isolated is substantially pure.
  • a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition may be isolated from a natural source (e.g, tissue) or from a source such as an engineered cell line.
  • tissue e.g., tissue
  • a source such as an engineered cell line.
  • substantially pure refers to material which is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.
  • subject refers to any animal (e.g ., a mammal), including, but not limited to, humans, non-human primates, canines, felines, rabbits, rodents, and the like.
  • pharmaceutically acceptable refers to a substance approved or approvable by a regulatory agency or listed in the U.S. Pharmacopeia, European Pharmacopeia, or other generally recognized pharmacopeia for use in animals, including humans.
  • pharmaceutically acceptable excipient, carrier, or adjuvant refers to an excipient, carrier, or adjuvant that can be administered to a subject (e.g., a human), together with at least one therapeutic agent and which is generally safe, non-toxic, and has no effect on the pharmacological activity of the therapeutic agent.
  • a pharmaceutically acceptable excipient, carrier, or adjuvant to be an inactive ingredient of any formulation.
  • pharmaceutical formulation or “pharmaceutical composition” as used herein refers to a preparation which is in such form as to permit the biological activity of the agent to be effective.
  • a pharmaceutical formulation or composition generally comprises additional components, such as a pharmaceutically acceptable excipient, carrier, adjuvant, buffers, etc.
  • an effective amount or “therapeutically effective amount” as used herein refers to the amount of an agent which is sufficient to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder or condition in a subject, and/or (ii) a symptom in a subject.
  • the term also encompasses an amount of an agent necessary for the (i) reduction or amelioration of the advancement or progression of a given disease, disorder, or condition, (ii) reduction or amelioration of the recurrence, development, or onset of a given disease, disorder, or condition, and/or (iii) the improvement or enhancement of the prophylactic or therapeutic effect(s) of another agent or therapy (e.g, an agent other than the binding agents provided herein).
  • therapeutic effect refers to the effect and/or ability of an agent to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder, or condition in a subject, and/or (ii) a symptom in a subject.
  • the term also encompasses the ability of an agent to (i) reduce or ameliorate the advancement or progression of a given disease, disorder, or condition, (ii) reduce or ameliorate the recurrence, development, or onset of a given disease, disorder, or condition, and/or (iii) to improve or enhance the prophylactic or therapeutic effect(s) of another agent or therapy (e.g an agent other than the binding agents provided herein).
  • treat or “treatment” or “treating” or “to treat” or “alleviate” or alleviation” or “alleviating” or “to alleviate” as used herein refers to therapeutic measures that aim to cure, slow down, lessen symptoms of, and/or halt progression of a pathologic condition or disorder.
  • prevent refers to the partial or total inhibition of the development, recurrence, onset, or spread of a disease, disorder, or condition, or a symptom thereof in a subject.
  • reference to “about” or “approximately” a value or parameter includes (and describes) embodiments that are directed to that value or parameter. For example, a description referring to “about X” includes description of “X”.
  • Amino acid (aa) sequences for human C3 (UniProtKB No. P01024) and cynomolgus monkey (“cyno”) are provided herein as SEQ ID NO: 1 and SEQ ID NO:7, respectively.
  • reference to amino acid positions of C3 refer to the numbering of amino acid sequences including the signal sequence.
  • the C3 -binding agent binds human C3 and has at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) of the following properties: (a) binds to cyno C3, (b) is an antagonist of C3, (c) inhibits C3 activity, (d) inhibits C3 cleavage, (e) inhibits C3 cleavage and C3a release, (f) inhibits complement activation, (g) inhibits activation of the alternative complement pathway, (h) inhibits activation of the classical complement pathway, (i) inhibits activation of the alternative complement pathway and inhibits activation of the classical complement pathway, (j) does not detectably bind to Factor Bb, (k) does not detectably bind to C3d, and (1) does not detectably bind to C3a.
  • the C3 -binding agent binds human C3 and has at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) of the following properties: (
  • the C3 -binding agent inhibits activation of the classical complement pathway. In some embodiments, the C3-binding agent inhibits activation of the alternative complement pathway and classical complement pathway. In some embodiments, a C3-binding agent binds C3. In some embodiments, a C3-binding agent binds a fragment of C3. In some embodiments, a C3- binding agent binds within a specific region of C3. In some embodiments, a C3-binding agent binds an epitope on C3. In some embodiments, a C3-binding agent binds a linear epitope on C3. In some embodiments, a C3-binding agent binds a conformational epitope on C3.
  • a C3-binding agent binds human C3. In some embodiments, a C3 -binding agent binds cyno C3. In some embodiments, a C3 -binding agent binds human C3 and cyno C3. In some embodiments, a C3-binding agent binds C3 and does not bind C3b. In some embodiments, a C3 -binding agent binds C3 and does not bind C3b at a detectable level. [00102] Assays for determining inhibition of C3 cleavage and C3a release are known to those skilled in the art.
  • the C3 -binding agent inhibits C3 cleavage and C3a release at a concentration of 0.1 to 1000 nM. In certain embodiments, a C3-binding agent inhibits C3 cleavage and C3a release if there is a reduction by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% in the amount of C3 cleavage and C3a release compared to C3 cleavage and C3a release in the absence of the C3-binding agent.
  • a C3-binding agent is determined to not detectably bind to factor Bb, C3d, or C3a if the C3 -binding agent does not bind to factor Bb, C3d, or C3a, respectively, at a concentration of up to 1 mM according to an assay described herein ( e.g ., BiaCore).
  • a C3-binding agent is an antibody.
  • the antibody is a recombinant antibody.
  • the antibody is a monoclonal antibody.
  • the antibody is a chimeric antibody.
  • the antibody is a humanized antibody.
  • the antibody is a human antibody.
  • the antibody is an IgA, IgD, IgE, IgG, or IgM antibody.
  • the antibody is an IgGl antibody.
  • the antibody is an IgG2 antibody.
  • the antibody is an IgG3 antibody.
  • the antibody is an IgG4 antibody.
  • the antibody is a human IgGl antibody. In some embodiments, the antibody is a human IgG2 antibody. In some embodiments, the antibody is a human IgG3 antibody. In some embodiments, the antibody is a human IgG4 antibody. In some embodiments, the antibody comprises a human kappa light chain constant region. In some embodiments, the antibody comprises a human lambda light chain constant region. In some embodiments, the antibody is an antibody fragment comprising an antigen-binding site. In some embodiments, the antibody is a scFv. In some embodiments, the antibody is a disulfide-linked scFv. In some embodiments, the antibody is a bispecific antibody or a multispecific antibody. In some embodiments, the antibody is a monovalent antibody. In some embodiments, the antibody is a monospecific antibody. In some embodiments, the antibody is a bivalent antibody.
  • the antibody is isolated. In some embodiments, the antibody is substantially pure.
  • a C3-binding agent is a polyclonal antibody.
  • Polyclonal antibodies can be prepared by any method known to those of skill in the art.
  • polyclonal antibodies are produced by immunizing an animal (e.g ., a rabbit, rat, mouse, goat, donkey) with an antigen of interest (e.g, a purified peptide fragment, a recombinant protein, or a fusion protein) using multiple subcutaneous or intraperitoneal injections.
  • an antigen of interest e.g, a purified peptide fragment, a recombinant protein, or a fusion protein
  • the antigen is conjugated to a carrier such as keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor.
  • KLH keyhole limpet hemocyanin
  • serum albumin serum albumin
  • bovine thyroglobulin or soybean trypsin inhibitor.
  • the antigen (with or without a carrier protein) is diluted in sterile saline and usually combined with an adjuvant (e.g, Complete or Incomplete Freund's Adjuvant) to form a stable emulsion.
  • an adjuvant e.g, Complete or Incomplete Freund's Adjuvant
  • polyclonal antibodies are recovered from the immunized animal (e.g, from blood or ascites).
  • the polyclonal antibodies are purified from serum or ascites according to standard methods in the art including, but not limited to, affinity chromatography, ion-exchange chromatography, gel electrophoresis, and/or dialysis.
  • a C3-binding agent is a monoclonal antibody.
  • Monoclonal antibodies can be prepared by any method known to those of skill in the art.
  • monoclonal antibodies are prepared using hybridoma methods known to one of skill in the art. For example, using a hybridoma method, a mouse, rat, rabbit, hamster, or other appropriate host animal, is immunized as described above.
  • lymphocytes are immunized in vitro.
  • the immunizing antigen is a human protein or a fragment thereof. In some embodiments, the immunizing antigen is a mouse protein or a fragment thereof.
  • lymphocytes are isolated and fused with a suitable myeloma cell line using, for example, polyethylene glycol.
  • the hybridoma cells are selected using specialized media as known in the art and unfused lymphocytes and myeloma cells do not survive the selection process.
  • Hybridomas that produce monoclonal antibodies directed specifically against a chosen antigen can be identified by a variety of methods including, but not limited to, immunoprecipitation, immunoblotting, and in vitro binding assays (e.g, flow cytometry, FACS, ELISA, SPR ( e.g. , Biacore), and radioimmunoassay).
  • the clones may be subcloned by limiting dilution techniques.
  • the hybridomas can be propagated either in in vitro culture using standard methods or in vivo as ascites tumors in an animal.
  • the monoclonal antibodies can be purified from the culture medium or ascites fluid according to standard methods in the art including, but not limited to, affinity chromatography, ion-exchange chromatography, gel electrophoresis, and dialysis.
  • C3-binding agents e.g., monoclonal antibodies
  • a polynucleotide or polynucleotides encoding an antibody are isolated from mature B-cells or hybridoma cells, such as by RT-PCR using oligonucleotide primers that specifically amplify the genes encoding the heavy and light chains of the antibody, and their sequence is determined using standard techniques.
  • the isolated polynucleotides encoding the heavy and light chains are then cloned into suitable expression vectors which produce the monoclonal antibodies when transfected into host cells such as E. coli , simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins.
  • a C3-binding agent is an antibody that is made using recombinant DNA techniques as known to one skilled in the art.
  • a polynucleotide or polynucleotides encoding the heavy and light chains thenare cloned into suitable expression vectors which produce the antibodies when transfected into host cells such as E. coli, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins.
  • recombinant antibodies are isolated from phage display libraries expressing variable domains or CDRs of a desired species. Screening of phage libraries can be accomplished by various techniques known in the art.
  • a C3-binding agent e.g., a monoclonal antibody
  • a C3-binding agent is modified by using recombinant DNA technology to generate alternative antibodies.
  • the constant domains of the light chain and heavy chain of a mouse monoclonal antibody are substituted for constant regions of a human antibody to generate a chimeric antibody.
  • the constant regions are truncated or removed to generate a desired antibody fragment of a monoclonal antibody.
  • site-directed or high-density mutagenesis of the variable region(s) is used to optimize specificity and affinity of a monoclonal antibody.
  • a C3-binding agent is a humanized antibody.
  • a humanized antibody comprises one or more amino acid residues that have been introduced into it from a source that is non-human.
  • humanization is performed by substituting one or more non-human CDR sequences for the corresponding CDR sequences of a human antibody.
  • the humanized antibodies are constructed by substituting all six CDRs of a non-human antibody (e.g., a mouse antibody) for the corresponding CDRs of a human antibody.
  • variable region framework for the humanized antibody.
  • variable region framework for the humanized antibody.
  • a particular variable region framework derived from a consensus sequence of all human antibodies of a particular subgroup of light or heavy chains is selected as the variable region framework.
  • variable region framework sequence is derived from the consensus sequences of the most abundant human subclasses.
  • human germline genes are used as the source of the variable region framework sequences.
  • Other methods for humanization include, but are not limited to, a method called “superhumanization” which is described as the direct transfer of CDRs to a human germline framework, a method termed Human String Content (HSC) which is based on a metric of “antibody humanness”, methods based on generation of large libraries of humanized variants (including phage, ribosomal, and yeast display libraries), and methods based on framework region shuffling.
  • HSC Human String Content
  • a C3-binding agent is a human antibody.
  • Human antibodies can be prepared using various techniques known in the art.
  • human antibodies are generated from immortalized human B lymphocytes immunized in vitro.
  • human antibodies are generated from lymphocytes isolated from an immunized individual. In any case, cells that produce an antibody directed against a target antigen can be generated and isolated.
  • a human antibody is selected from a phage library, where that phage library expresses human antibodies.
  • phage display technology may be used to produce human antibodies and antibody fragments in vitro , from immunoglobulin variable region gene repertoires from unimmunized donors.
  • human antibodies are produced in transgenic mice that contain human immunoglobulin loci. Upon immunization these mice are capable of producing the full repertoire of human antibodies in the absence of endogenous immunoglobulin production.
  • a C3 -binding agent is an antibody fragment.
  • antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, single chain antibody molecules, scFv, disulfide-linked scFv (dsscFv), nanobodies, diabodies, tribodies, tetrabodies, minibodies, dual variable domain antibodies (DVD), single variable domain antibodies (e.g ., camelid antibodies), and multispecific antibodies comprising antibody fragments.
  • a C3-binding agent is an scFv antibody.
  • ScFv antibodies are molecules that comprise a variable heavy chain region and a variable light chain region linked to form a single polypeptide.
  • a scFv comprises a disulfide bond formed between the heavy chain variable region and the light chain variable region.
  • a scFv antibody comprises a disulfide bond that increases stability of the scFv molecule.
  • a scFv antibody comprises a disulfide bond that increases thermostability of the scFv molecule.
  • ScFv antibodies can be produced using recombinant technologies known in the art.
  • Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody.
  • the antibody fragments described herein can be produced using recombinant technologies known in the art (e.g., E.coli or phage expression).
  • a C3-binding agent is a bispecific antibody.
  • Bispecific antibodies are capable of recognizing and binding at least two different antigens or epitopes.
  • the different epitopes can either be within the same molecule (e.g, two epitopes on C3) or on different molecules (e.g, one epitope on C3 and one epitope on a different target).
  • a bispecific antibody has enhanced potency as compared to an individual antibody or to a combination of more than one antibody.
  • a bispecific antibody has reduced toxicity as compared to an individual antibody or to a combination of more than one antibody.
  • a bispecific antibody has the ability to synchronize the PK of two active binding agents wherein the two individual binding agents have different PK profiles.
  • a bispecific antibody has the ability to concentrate the actions of two agents in a common area (e.g, tissue) in a subject.
  • a bispecific antibody has the ability to concentrate the actions of two agents to a common target (e.g ., a specific cell type).
  • a bispecific antibody has the ability to target the actions of two agents to more than one biological pathway or function.
  • a bispecific antibody has the ability to target two different cells and bring them closer together. [00122] In some embodiments, a bispecific antibody has decreased toxicity and/or side effects. In some embodiments, a bispecific antibody has decreased toxicity and/or side effects as compared to a mixture of the two individual antibodies or the antibodies as single agents. In some embodiments, a bispecific antibody has an increased therapeutic index. In some embodiments, a bispecific antibody has an increased therapeutic index as compared to a mixture of the two individual antibodies or the antibodies as single agents.
  • the bispecific antibodies comprise heavy chain constant regions with modifications in the amino acids that are part of the interface between the two heavy chains. These modifications are made to enhance heterodimer formation and generally reduce or eliminate homodimer formation.
  • the bispecific antibodies are generated using a knobs-into-holes (KIH) strategy.
  • the bispecific antibodies comprise variant hinge regions incapable of forming disulfide linkages between identical heavy chains (e.g., reduce homodimer formation).
  • the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered electrostatic interactions.
  • the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered hydrophobic/hydrophilic interactions.
  • a bispecific antibody is an intact/full length antibody.
  • a bispecific antibody comprises antibody fragments comprising two antigen-binding sites.
  • C3-binding agents with more than two valencies are also contemplated. In some embodiments, trispecific or tetraspecific antibodies are generated.
  • a C3-binding agent is an antibody that binds C3. In some embodiments, an anti-C3 antibody binds human C3. In some embodiments, an anti-C3 antibody binds cyno C3. In some embodiments, an anti-C3 antibody binds human C3 and cyno C3. In some embodiments, an anti-C3 antibody does not bind rat C3. In some embodiments, an anti-C3 antibody binds a C3 epitope.
  • an anti-C3 antibody does not bind C3a, C3b, C3c, C3d, or iC3b. In some embodiments, an anti-C3 antibody does not bind C3a, C3b, C3c, C3d, and iC3b. In some embodiments, an anti-C3 antibody does not bind C3a. In some embodiments, an anti-C3 antibody does not bind C3c. In some embodiments, an anti-C3 antibody does not bind C3d. In some embodiments, an anti-C3 antibody does not bind iC3b. In some embodiments, an anti-C3 antibody does not bind C3b.
  • an anti- C3 antibody does not bind C3a, C3b, C3c, C3d, or iC3b at a detectable level. In some embodiments, an anti-C3 antibody binds C3 with an affinity that is at least 20-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b. In some embodiments, an anti-C3 antibody binds C3 with an affinity that is at least 50-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b.
  • an anti-C3 antibody binds C3 with an affinity that is at least 100-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b. In some embodiments, an anti-C3 antibody binds C3 and does not bind C3b at a detectable level.
  • a C3-binding agent is a variant of an anti-C3 antibody described herein. A variant of an anti-C3 antibody described herein must still bind C3.
  • a variant of an anti-C3 antibody described herein comprises one to thirty amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to twenty-five amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to twenty amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to fifteen amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to ten amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to five amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to three amino acid substitutions. In some embodiments, the amino acid substitution(s) is in a CDR of the antibody. In some embodiments, the amino acid substitution(s) is not in a CDR of the antibody. In some embodiments, the amino acid substitution(s) is in a framework region of the antibody.
  • the amino acid substitution(s) is a conservative amino acid substitution.
  • CDRs of an antibody are defined by a variety of methods/sy stems by those skilled in the art. These systems and/or definitions have been developed and refined over a number of years and include Rabat, Chothia, IMGT, AbM, and Contact.
  • the Rabat definition is based on sequence variability and generally is the most commonly used.
  • the Chothia definition is based on the location of the structural loop regions.
  • the IMGT system is based on sequence variability and location within the structure of the variable domain.
  • the AbM definition is a compromise between Rabat and Chothia.
  • the Contact definition is based on analyses of the available antibody crystal structures.
  • An Exemplary system is a combination of Rabat and Chothia.
  • Software programs e.g ., abYsis) are available and known to those of skill in the art for analysis of antibody sequences and determination of CDRs.
  • the specific CDR sequences defined herein are generally based on a combination of Rabat and Chothia definitions (Exemplary system). However, it will be understood that reference to a heavy chain CDR or CDRs and/or a light chain CDR or CDRs of a specific antibody will encompass all CDR definitions as known to those of skill in the art.
  • a C3-binding agent comprises one, two, three, four, five, and/or six CDRs of any one of the antibodies described herein.
  • an anti-C3 antibody comprises one, two, three, four, five, and/or six CDRs of any one of the antibodies described herein.
  • Anti-C3 antibodies have been described in U.S. Patent Application No.
  • 2019/0322730 including antibodies 38G10, Hz38G10, Hz38G10(G56A), Hz38G10(G56T), Hz38G10(N55E), Hz38G10(N55Q), 3D8, 3G8, 1502, 27A8, 28C3, 38F5, 62B11, 62F2, and 63 A3 described herein.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 1 A, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 1A.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table IB, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table IB.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 1C, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 1C.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 2, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 2.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 3, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 3.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 4, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 4.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 5, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 5.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 6, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 6.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 7, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 7.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 8, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 8.
  • a C3 -binding agent comprises (i) a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 9, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 9.
  • a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 10, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 10.
  • a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from an antibody described herein. In some embodiments, a C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from an antibody described herein. In some embodiments, a C3-binding agent comprises a humanized version or humanized variant of an antibody described herein.
  • a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from antibody 38G10 or a humanized version thereof.
  • a C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10 or variants thereof (e.g., HzG38G10(G56A), HzG38G10(G56T), HzG38G10(N55E), and HzG38G10(N55Q)).
  • a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody 38G10. In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10. In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(G56A).
  • a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(G56T). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(N55E). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(N55Q).
  • a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 using any of the CDR definitions exemplified in Table 1A, Table IB, Table 1C, or Tables 2-10.
  • a C3-binding agent comprises one or more of the following CDRs: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHN GGTT YN QNF T G (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a heavy chain CDR1 comprising the amino acid sequence GYTFTDF
  • a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3- binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprises a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3- binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence GYTFTDF (
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO: 9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO:l 1), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14
  • a C3- binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3- binding agent comprises a light chain variable region comprising light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent is a variant of an agent described herein.
  • a C3-binding agent can comprise a heavy chain variable region comprising a heavy chain CDR1 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR1 described herein; a heavy chain CDR2 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR2 described herein; a heavy chain CDR3 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR3 described herein; a light chain CDR1 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR1 described herein; a light chain CDR2 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR2 described herein; and a light chain CDR3 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR3 described herein.
  • a C3 -binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9) or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10) or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), or a variant thereof comprising
  • a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification reduces deamidation.
  • a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to reduce deamidation within the CDR sequence.
  • Deamidation is a chemical reaction in which an amide functional group in the side chain of the amino acids asparagine (Asn or N) or glutamine (Gin or Q) is removed or converted to another functional group.
  • asparagine is converted to aspartic acid or isoaspartic acid and glutamine is converted to glutamic acid or polyglutamic acid.
  • deamidation may change the structure, function, and/or stability of a polypeptide, potentially resulting in decreased biological activity.
  • a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification reduces isomerization.
  • a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to reduce isomerization. Isomerization is a chemical process by which a compound is transformed into any of its isomeric forms, i.e., forms with the same chemical composition but with different structure or configuration and, potentially with different physical and chemical properties. Studies have shown that aspartate (Asp or D) isomerization within a CDR can impact antibody binding and/or stability.
  • a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification eliminates a glycosylation site.
  • a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to eliminate a glycosylation site.
  • the consensus glycosylation site for N-linked glycans is N-X-S/T, wherein X can be any amino acid except proline.
  • a glycosylation site within a variable region and/or within a CDR will impact antibody structure, binding, and/or stability.
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3- binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprise a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3- binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11).
  • a C3-binding agent comprise a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, or SEQ ID NO:224; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32 or SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 31.
  • a C3- binding agent comprises a light chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:33.
  • a C3 -binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:34. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:222.
  • a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:223. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:224.
  • a C3-binding agent comprises a light chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31 and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 31 and a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31 and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 31 and a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 31 and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:31 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:34 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:32.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence SEQ ID NO:35.
  • a C3 -binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:222, SEQ ID NO:223 or SEQ ID NO:224. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:222 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
  • a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:223 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:224 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:78.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:78.
  • a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:78.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:96.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:96.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:96.
  • a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:95.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:96.
  • a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:95 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:96.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 113; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 113; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:l 13; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 113.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 113 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 114.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 131.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 131 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 132.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 149.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 149 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168.
  • a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 167.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 167 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 185.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 185 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:203.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:203 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:204.
  • a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221.
  • a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221.
  • a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:220.
  • a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:220 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 221.
  • a C3-binding agent described herein comprises an antibody in which at least one or more of the constant regions has been modified or deleted.
  • the antibodies comprise at least one modification in the hinge region of the heavy chain.
  • the antibodies comprise modifications to one or more of the three heavy chain constant regions (CHI, CH2 or CH3) and/or to the light chain constant region (CL).
  • the heavy chain constant region of the modified antibodies comprises at least one human constant region.
  • the heavy chain constant region of the modified antibodies comprises more than one human constant region.
  • modifications to the constant region comprise additions, deletions, or substitutions of one or more amino acids in one or more regions.
  • one or more regions are partially or entirely deleted from the constant regions of the modified antibodies.
  • the entire CH2 domain has been removed from an antibody ( ⁇ CH2 constructs).
  • a deleted constant region is replaced by a short amino acid spacer that provides some of the molecular flexibility typically imparted by the absent constant region.
  • a modified antibody comprises a CH3 domain directly fused to the hinge region of the antibody.
  • a modified antibody comprises a peptide spacer inserted between the hinge region and modified CH2 and/or CH3 domains.
  • the constant region(s) of an antibody mediates several effector functions and these effector functions can vary depending on the isotype of the antibody.
  • binding of the Cl component of complement to the Fc region of IgG or IgM antibodies (bound to antigen) activates the complement system.
  • Activation of complement is important in the opsonization and lysis of cell pathogens.
  • the activation of complement also stimulates the inflammatory response and can be involved in autoimmune hypersensitivity.
  • the Fc region of an antibody can bind a cell expressing a Fc receptor (FcR).
  • Fc receptors which are specific for different classes of antibody, including IgG (gamma receptors), IgE (epsilon receptors), IgA (alpha receptors) and IgM (mu receptors). Binding of antibody to Fc receptors on cell surfaces triggers a number of important and diverse biological responses including engulfment and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target cells by killer cells (called antibody-dependent cell cytotoxicity or ADCC), release of inflammatory mediators, placental transfer, and control of immunoglobulin production.
  • IgG gamma receptors
  • IgE epsilon receptors
  • IgA alpha receptors
  • IgM mi receptors
  • an antibody comprises a variant Fc region.
  • the amino acid sequences of the Fc region of human IgGl, IgG2, IgG3, and IgG4 are known to those of ordinary skill in the art (e.g ., a representative human IgGl is SEQ ID NO:42).
  • Fc regions with amino acid variations have been identified in native antibodies.
  • a variant Fc region is engineered with substitutions at specific amino acid positions as compared to a native Fc region (e.g ., SEQ ID NOs:43-46).
  • the modified antibodies provide for altered effector functions that, in turn, affect the biological profile of the antibody.
  • the deletion or inactivation (through point mutations or other means) of a constant region reduces Fc receptor binding of the modified antibody as it circulates.
  • the constant region modifications increase the serum half-life of the antibody.
  • the constant region modifications reduce the serum half-life of the antibody.
  • the constant region modifications decrease or remove ADCC and/or complement dependent cytotoxicity (CDC) of the antibody.
  • specific amino acid substitutions in a human IgGl Fc region with corresponding IgG2 or IgG4 residues reduce effector functions (e.g., ADCC and CDC) in the modified antibody.
  • an antibody does not have one or more effector functions.
  • the antibody has no ADCC activity and/or no CDC activity.
  • the antibody does not bind an Fc receptor and/or complement factors.
  • the antibody has no effector function(s) (e.g, “effectorless” antibodies).
  • the constant region modifications increase or enhance ADCC and/or CDC of the antibody.
  • the constant region is modified to eliminate disulfide linkages or oligosaccharide moieties. In some embodiments, the constant region is modified to add/substitute one or more amino acids to provide one or more cytotoxin, oligosaccharide, or carbohydrate attachment sites.
  • a C3-binding agent comprises (a) a heavy chain comprising a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 using any one of the CDR definitions as shown in Tables 1 A- 1C, and (b) a light chain comprising a light chain CDR1, a light chain CDR2, and a light chain CDR3 using any of the CDR definitions as shown in Tables 1 A-1C, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:41.
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14), wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO: 10, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:9,
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:25, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO: 39,
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:27, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:48, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO: 39,
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:52, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:
  • a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:56, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO: 39,
  • a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39. In some embodiments, a C3-binding agent is an antibody that comprises a light chain of SEQ ID NO:41.
  • a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41.
  • antibody variants are prepared by introducing appropriate nucleotide changes into the encoding DNA, and/or by synthesis of the desired antibody or polypeptide. Using these antibody variants it may be possible to disrupt the activity or effector function provided by a specific sequence or region while substantially maintaining the structure, binding activity, and other desired characteristics of the modified antibody.
  • the present disclosure further embraces additional variants and equivalents that are substantially homologous to the recombinant, monoclonal, chimeric, humanized, and human antibodies, or antibody fragments thereof, described herein.
  • it is desirable to modulate biological properties of the antibody including but not limited to, specificity, thermostability, expression level, effector function(s), glycosylation, immunogenicity, or solubility.
  • amino acid changes may alter post-translational processes of an antibody, such as changing the number or position of glycosylation sites or altering membrane anchoring characteristics.
  • Variations may be a substitution, deletion, or insertion of one or more nucleotides encoding the antibody or polypeptide that results in a change in the amino acid sequence as compared with the native antibody or polypeptide sequence.
  • amino acid substitutions are the result of replacing one amino acid with another amino acid having similar structural and/or chemical properties, such as the replacement of a leucine with a serine, e.g., conservative amino acid substitutions.
  • Insertions or deletions may optionally be in the range of about 1 to 5 amino acids.
  • the substitution, deletion, or insertion includes less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the parent molecule.
  • variations in the amino acid sequence that are biologically useful and/or relevant are determined by systematically making insertions, deletions, or substitutions in the sequence and testing the resulting variant proteins for activity as compared to the parental antibody.
  • variants may include addition of amino acid residues at the amino- and/or carboxyl-terminal end of the antibody or polypeptide.
  • the length of additional amino acids residues may range from one residue to a hundred or more residues.
  • a variant comprises an N-terminal methionyl residue.
  • a variant does not comprise an N-terminal methionyl residue.
  • the variant comprises an additional polypeptide/protein, /. e. , a fusion protein.
  • a variant is engineered to be detectable and may comprise a detectable label and/or protein (e.g, an enzyme).
  • a cysteine residue not involved in maintaining the proper conformation of an antibody may be substituted or deleted to modulate the antibody’s characteristics, for example, to improve oxidative stability and/or prevent aberrant disulfide crosslinking.
  • one or more cysteine residues may be added to create disulfide bond(s) to improve stability.
  • an antibody of the present disclosure is “deimmunized”.
  • the deimmunization of antibodies generally consists of introducing specific amino acid mutations (e.g., substitutions, deletions, additions) that result in removal of T-cell epitopes without significantly reducing the binding affinity or other desired activities of the antibody.
  • the variant antibodies or polypeptides described herein may be generated using methods known in the art, including but not limited to, site-directed mutagenesis, alanine scanning mutagenesis, and PCR mutagenesis.
  • C3-binding agents described herein are chemically modified.
  • the C3 -binding agents are anti-C3 antibodies that have been chemically modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, and/or linkage to a cellular ligand or other protein. Any of numerous chemical modifications may be carried out by known techniques.
  • a C3-binding agent is attached, either directly or indirectly, to a half-life extending moiety including, but not limited to, a Fc region, a CH3 domain of an immunoglobulin, polyethylene glycol (PEG), a PEG mimetic, XTEN®, serum albumin, polysialic acid, N-(2-hydroxypropyl)methacrylamide, or dextran.
  • a half-life extending moiety including, but not limited to, a Fc region, a CH3 domain of an immunoglobulin, polyethylene glycol (PEG), a PEG mimetic, XTEN®, serum albumin, polysialic acid, N-(2-hydroxypropyl)methacrylamide, or dextran.
  • an anti-C3 antibody described herein is attached, either directly or indirectly, to a half-life extending moiety including, but not limited to, polyethylene glycol (PEG), a PEG mimetic, XTEN®, serum albumin, polysialic acid, N-(2-hydroxypropyl)methacrylamide, or dextran.
  • PEG polyethylene glycol
  • XTEN® XTEN®
  • serum albumin serum albumin
  • polysialic acid polysialic acid
  • N-(2-hydroxypropyl)methacrylamide or dextran.
  • the present disclosure encompasses C3-binding agents built upon nonimmunoglobulin backbones, wherein the agents bind to the same epitope or essentially the same epitope as an anti-C3 antibody disclosed herein.
  • a non- immunoglobulin-based binding agent is an agent that competes with an anti-C3 antibody described herein in a competitive binding assay.
  • alternative C3-binding agents comprise a scaffold protein.
  • scaffold proteins can be assigned to one of three groups based on the architecture of their backbone (1) scaffolds consisting of a-helices; (2) small scaffolds with few secondary structures or an irregular architecture of a-helices and b-sheets; and (3) scaffolds consisting of predominantly b-sheets.
  • Scaffold proteins include, but are not limited to, anticalins, which are based upon the lipocalin scaffold; adnectins, which are based on the 10 th domain of human fibronectin type 3; affibodies, which are based on the B-domain in the Ig-binding region of Staphylococcus aureus protein A; darpins, which are based on ankyrin repeat domain proteins; fynomers, which are based on the SH3 domain of the human Fyn protein kinase; affitins, which are based on Sac7d from Sulfolobus acidocaldarius ; affilins, which are based on human g-B-crystallin or human ubiquitin; avimers, which are based on the A-domains of membrane receptor proteins; knottins (cysteine knot miniproteins), which are based upon a stable 30-amino acid anti -parallel b-strand protein fold; and Kun
  • a C3-binding agent comprises an engineered scaffold protein comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3 shown in Tables 1A-1C or Tables 2-10.
  • a C3-binding agent comprises an engineered scaffold protein comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10),
  • YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • antigen-antibody interactions are non-covalent and reversible, formed by a combination of hydrogen bonds, hydrophobic interactions, electrostatic and van der Waals forces.
  • affinity and/or avidity are commonly used mentioned.
  • the binding of an antibody to its antigen is a reversible process, and the affinity of the binding is typically reported as an equilibrium dissociation constant (KD).
  • KD is the ratio of an antibody dissociation rate (k 0ff ) (how quickly it dissociates from its antigen) to the antibody association rate (k 0n ) (how quickly it binds to its antigen).
  • KD values are determined by measuring the k 0n and k 0ff rates of a specific antibody/antigen interaction and then using a ratio of these values to calculate the KD value.
  • KD values may be used to evaluate and rank order the strength of individual antibody/antigen interactions. The lower the KD of an antibody, the higher the affinity of the antibody for its target.
  • affinity is measured using SPR technology in a Biacore system. Avidity gives a measure of the overall strength of an antibody-antigen complex. It is dependent on three major parameters: (i) affinity of the antibody for the target, (ii) valency of both the antibody and antigen, and (iii) structural arrangement of the parts that interact.
  • a C3 -binding agent binds C3 with a dissociation constant (KD) of about 1 mM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, about 0.1 nM or less, 50 pM or less, 10 pM or less, or 1 pM or less.
  • KD dissociation constant
  • a C3 -binding agent binds C3 with a KD of about 20 nM or less.
  • a C3-binding agent binds C3 with a KD of about 10 nM or less.
  • a C3-binding agent binds C3 with a KD of about 1 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 0.5 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 0.1 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 50 pM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 25 pM or less.
  • a C3-binding agent binds C3 with a KD of about 10 pM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 1 pM or less.
  • the dissociation constant of the binding agent for C3 is the dissociation constant determined using a C3 protein immobilized on a Biacore chip and the binding agent flowed over the chip. In some embodiments, the dissociation constant of the binding agent for C3 is the dissociation constant determined using the binding agent captured by an anti-human IgG antibody on a Biacore chip and soluble C3 flowed over the chip.
  • a C3 -binding agent binds C3 with a half maximal effective concentration (EC50) of about 1 pM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 nM or less. In some embodiments, a C3 -binding agent binds to human C3 with an EC50 of about 1 pM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 nM or less.
  • EC50 half maximal effective concentration
  • a C3-binding agent binds cyno C3 and/or human C3 with an EC50 of about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less or about 0.1 nM or less.
  • the C3 -binding agents described herein can be produced by any suitable method known in the art. Such methods range from direct protein synthesis methods to constructing a DNA sequence encoding polypeptide sequences and expressing those sequences in a suitable host.
  • a DNA sequence is constructed using recombinant technology by isolating or synthesizing a DNA sequence encoding a wild-type protein of interest.
  • the sequence can be mutagenized by site- specific mutagenesis to provide functional variants thereof.
  • a DNA sequence encoding a polypeptide of interest is constructed by chemical synthesis using an oligonucleotide synthesizer.
  • Oligonucleotides can be designed based on the amino acid sequence of the desired polypeptide and selecting those codons that are favored in the host cell in which the recombinant polypeptide of interest will be produced. Standard methods can be applied to synthesize a polynucleotide sequence encoding an isolated polypeptide of interest. For example, a complete amino acid sequence can be used to construct a back-translated gene. Further, a DNA oligomer containing a nucleotide sequence coding for the particular isolated polypeptide can be synthesized. For example, several small oligonucleotides coding for portions of the desired polypeptide can be synthesized and then ligated.
  • the individual oligonucleotides typically contain 5' or 3' overhangs for complementary assembly.
  • the polynucleotide sequences encoding a particular polypeptide of interest can be inserted into an expression vector and operatively linked to an expression control sequence appropriate for expression of the protein in a desired host. Proper assembly can be confirmed by nucleotide sequencing, restriction enzyme mapping, and/or expression of a biologically active polypeptide in a suitable host.
  • the gene in order to obtain high expression levels of a transfected gene in a host, the gene must be operatively linked to transcriptional and translational expression control sequences that are functional in the chosen expression host.
  • recombinant expression vectors are used to amplify and express DNA encoding antibodies, or fragments thereof, against human C3.
  • recombinant expression vectors can be replicable DNA constructs which have synthetic or cDNA-derived DNA fragments encoding a polypeptide chain of a C3- binding agent, such as an anti-C3 antibody, or antigen-binding fragment thereof, operatively linked to suitable transcriptional and/or translational regulatory elements derived from mammalian, microbial, viral or insect genes.
  • a transcriptional unit generally comprises an assembly of (1) a genetic element or elements having a regulatory role in gene expression, for example, transcriptional promoters or enhancers, (2) a structural or coding sequence which is transcribed into mRNA and translated into protein, and (3) appropriate transcription and translation initiation and termination sequences. Regulatory elements can include an operator sequence to control transcription. The ability to replicate in a host, usually conferred by an origin of replication, and a selection gene to facilitate recognition of transformants can additionally be incorporated. DNA regions are “operatively linked” when they are functionally related to each other.
  • DNA for a signal peptide is operatively linked to DNA for a polypeptide if it is expressed as a precursor which participates in the secretion of the polypeptide; a promoter is operatively linked to a coding sequence if it controls the transcription of the sequence; or a ribosome binding site is operatively linked to a coding sequence if it is positioned so as to permit translation.
  • structural elements intended for use in yeast expression systems include a leader sequence enabling extracellular secretion of translated protein by a host cell.
  • a polypeptide may include an N-terminal methionine residue.
  • an expression control sequence and an expression vector generally depends upon the choice of host.
  • a wide variety of expression host/vector combinations can be employed.
  • Useful expression vectors for eukaryotic hosts include, for example, vectors comprising expression control sequences from SV40, bovine papilloma virus, adenovirus, and cytomegalovirus.
  • Useful expression vectors for bacterial hosts include known bacterial plasmids, such as plasmids from E. coli , including pCRl, pBR322, pMB9 and their derivatives, and wider host range plasmids, such as Ml 3 and other filamentous single-stranded DNA phages.
  • the C3-binding agents of the present disclosure can be expressed from one or more vectors.
  • a heavy chain polypeptide is expressed by one vector and a light chain polypeptide is expressed by a second vector.
  • a heavy chain polypeptide and a light chain polypeptide are expressed by one vector.
  • Suitable host cells for expression of a C3-binding agent or a C3 protein or fragment thereof to use as an antigen or immunogen include prokaryotes, yeast cells, insect cells, or higher eukaryotic cells under the control of appropriate promoters.
  • Prokaryotes include gram-negative or gram-positive organisms, for example E. coli or Bacillus.
  • Higher eukaryotic cells include established cell lines of mammalian origin as described herein. Cell-free translation systems may also be employed.
  • Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cellular hosts, as well as methods of protein production, including antibody production are well known in the art.
  • Suitable mammalian host cell lines include, but are not limited to, COS-7 (monkey kidney-derived), L-929 (murine fibroblast-derived),
  • Mammalian expression vectors can comprise non-transcribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, and other 5' or 3' flanking non-transcribed sequences, and 5' or 3' non-translated sequences, such as necessary ribosome binding sites, a polyadenylation site, splice donor and acceptor sites, and transcriptional termination sequences.
  • Expression of recombinant proteins in insect cell culture systems e.g ., baculovirus also offers a robust method for producing correctly folded and biologically functional proteins.
  • Baculovirus systems for production of heterologous proteins in insect cells are well-known to those of skill in the art.
  • the present disclosure provides cells comprising the C3-binding agents described herein.
  • the cells produce the C3-binding agents described herein.
  • the cells produce an antibody.
  • the cells produce an antibody that binds human C3.
  • the cells produce an antibody that binds cyno C3.
  • the cells produce an antibody that binds human C3 and cyno C3.
  • the cells produce an antibody designated 38G10.
  • the cells produce a humanized version of antibody 38G10, referred to as Hz38G10.
  • the cells produce a variant of Hz38G10.
  • the cells produce antibody Hz38G10(G56A).
  • the cell is a prokaryotic cell.
  • the cell is an eukaryotic cell.
  • the cell is a mammalian cell.
  • the cell is a hybridoma cell.
  • Proteins produced by a host cell can be purified according to any suitable method.
  • Standard methods include chromatography (e.g., ion exchange, affinity, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for protein purification.
  • Affinity tags such as hexa-histidine (SEQ ID NO:47), maltose binding domain, influenza coat sequence, and glutathione-S- transferase can be attached to the protein to allow easy purification by passage over an appropriate affinity column.
  • Affinity chromatography used for purifying immunoglobulins can include Protein A, Protein G, and Protein L chromatography.
  • Isolated proteins can be physically characterized using such techniques as proteolysis, size exclusion chromatography (SEC), mass spectrometry (MS), nuclear magnetic resonance (NMR), isoelectric focusing (IEF), high performance liquid chromatography (HPLC), and x-ray crystallography.
  • SEC size exclusion chromatography
  • MS mass spectrometry
  • NMR nuclear magnetic resonance
  • IEF isoelectric focusing
  • HPLC high performance liquid chromatography
  • x-ray crystallography x-ray crystallography
  • supernatants from expression systems which secrete recombinant protein into culture media are first concentrated using a commercially available protein concentration filter, for example, an Amicon® or Millipore Pellicon® ultrafiltration unit. Following the concentration step, the concentrate can be applied to a suitable purification matrix.
  • a suitable purification matrix for example, an anion exchange resin is employed, for example, a matrix or substrate having pendant diethylaminoethyl (DEAE) groups.
  • the matrices can be acrylamide, agarose, dextran, cellulose, or other types commonly employed in protein purification.
  • a cation exchange step is employed.
  • Suitable cation exchangers include various insoluble matrices comprising sulfopropyl or carboxymethyl groups.
  • a hydroxyapatite media is employed, including but not limited to, ceramic hydroxyapatite (CHT).
  • CHT ceramic hydroxyapatite
  • one or more reverse-phase HPLC steps employing hydrophobic RP-HPLC media, e.g, silica gel having pendant methyl or other aliphatic groups, are employed to further purify a recombinant protein.
  • hydrophobic interaction chromatography HIC is used to separate recombinant proteins based on their hydrophobicity.
  • HIC is a useful separation technique for purifying proteins while maintaining biological activity due to the use of conditions and matrices that operate under less denaturing conditions than some other techniques.
  • Anti-C3 antibodies of the present disclosure may be analyzed for their physical/chemical properties and/or biological activities by various assays known in the art.
  • an anti-C3 antibody is tested for its ability to bind C3 (e.g, human C3 and/or cyno C3). Binding assays include, but are not limited to, SPR (e.g, Biacore), ELISA, and FACS.
  • an anti-C3 antibody is tested for its ability to inhibit, reduce, or block complement activity.
  • an anti-C3 antibody is tested for its ability to inhibit, reduce, or block C3 activity.
  • Assays include, but are not limited to, hemolysis assays and C3a release assays.
  • antibodies may be evaluated for solubility, stability, thermostability, viscosity, expression levels, expression quality, and/or purification efficiency.
  • monoclonal antibodies generated against C3 are grouped based upon the epitope each individual antibody recognizes, a process known as “epitope binning”.
  • epitope binning antibodies are tested in a pairwise combinatorial manner and antibodies that compete with each other are grouped together into bins.
  • a premix binning assay a first antibody is immobilized on a surface and a premixed solution of a second antibody and antigen is flowed over the immobilized first antibody.
  • the antigen is immobilized on a surface and the two antibodies are flowed over the immobilized antigen and compete to bind.
  • antibodies that block one another can be identified.
  • a competitive blocking profile is created for each antibody relative to the other antibodies.
  • the blocking results determine which bin each antibody is placed in.
  • High-throughput methods of epitope binning are known in the art and allow for screening and characterization of large numbers of antibodies within a short period of time.
  • Antibodies that bind similar epitopes often share similar functions and/or capabilities. Conversely, antibodies that bind different epitopes may have different functional activities.
  • Epitope mapping is the process of identifying the binding site, or epitope on a target protein/antigen where an antibody (or other binding agent) binds.
  • a variety of methods are known in the art for mapping epitopes on target proteins. These methods include mutagenesis, including but not limited to, shotgun mutagenesis, site-directed mutagenesis, and alanine scanning; domain or fragment scanning, peptide scanning ( e.g ., Pepscan technology); display methods (e.g, phage display, microbial display, and ribosome/mRNA display); methods involving proteolysis and mass spectroscopy; and structural determination (e.g, x-ray crystallography and NMR).
  • purified anti-C3 antibodies are characterized by assays including, but not limited to, N-terminal sequencing, amino acid analysis, high pressure liquid chromatography (HPLC), mass spectrometry, ion exchange chromatography, and papain digestion.
  • assays including, but not limited to, N-terminal sequencing, amino acid analysis, high pressure liquid chromatography (HPLC), mass spectrometry, ion exchange chromatography, and papain digestion.
  • assays are provided for identifying anti-C3 antibodies that affect C3 activity.
  • hemolysis assays are used to assess the functional activity of the complement system. Hemolysis assays have been modified over the years to assess the activity of the different complement pathways and/or individual complement components of the cascade. In some embodiments, a hemolysis assay is used to assess activity of the alternative pathway. In some embodiments, a hemolysis assay is used to assess activity of the classical pathway.
  • a C3 -binding agent binds C3 and inhibits C3 activation of the alternative pathway. In some embodiments, a C3-binding agent binds C3 and inhibits activation of the alternative pathway, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3-binding agent inhibits activation of the alternative pathway by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%. In some embodiments, percent inhibition is used to calculate an ICso (half maximal inhibitory concentration) for the C3 -binding agent.
  • a C3 -binding agent that inhibits activation of the alternative pathway is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
  • a C3 -binding agent described herein binds C3 and inhibits C3 activation of the classical pathway. In some embodiments, a C3-binding agent binds C3 and inhibits activation of the classical pathway, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3-binding agent inhibits activation of the classical pathway by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%.
  • a C3-binding agent that inhibits activation of the classical pathway is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
  • a C3 -binding agent described herein binds C3 and inhibits C3 activation of the classical and alternative pathways. In some embodiments, a C3 -binding agent binds C3 and inhibits activation of the classical and alternative pathways, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3 -binding agent inhibits activation of the classical and alternative pathways by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%.
  • a C3 -binding agent that inhibits activation of the classical and alternative pathways is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
  • an immunochemical assay to determine the presence and/or amount of individual components e.g ., C3a
  • a C3-binding agent e.g., an antibody
  • the present disclosure also provides conjugates comprising an anti-C3 antibody described herein.
  • the antibody is attached to a second molecule.
  • the antibody is conjugated to a cytotoxic agent or moiety.
  • the antibody is conjugated to a cytotoxic agent to form an ADC (antibody-drug conjugate).
  • the cytotoxic agent is a chemotherapeutic agent including, but not limited to, methotrexate, adriamycin/doxorubicin, melphalan, mitomycin C, chlorambucil, duocarmycin, daunorubicin, pyrrolobenzodiazepines (PBDs), or other intercalating agents.
  • the cytotoxic agent is a microtubule inhibitor including, but not limited to, auristatins, maytansinoids (e.g., DM1 and DM4), and tubulysins.
  • the cytotoxic agent is an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof, including, but not limited to, diphtheria A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tric
  • an antibody is conjugated to one or more small molecule toxins, such as calicheamicins, maytansinoids, trichothenes, and CC1065.
  • a derivative of any one of these toxins may be used as long as the derivative retains the cytotoxic activity of the parent molecule.
  • Conjugates comprising an anti-C3 antibody described herein may be made using any suitable method known in the art.
  • conjugates are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3- (2-pyridyidithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HC1), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis(p- azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p- diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as l,5-difluoro-2, 4-dinitrobenzene).
  • SPDP N-succinimidyl-3- (2-pyridyidi
  • an anti-C3 antibody described herein is conjugated to a detectable substance or molecule that allows the antibody to be used for diagnosis and/or detection.
  • a detectable substance can include but is not limited to, enzymes, such as horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and acetylcholinesterase; prosthetic groups, such as biotin and flavine(s); fluorescent materials, such as, umbelliferone, fluorescein, fluorescein isothiocyanate (FITC), rhodamine, tetramethylrhodamine isothiocyanate (TRITC), dichlorotriazinylamine fluorescein, dansyl chloride, cyanine (Cy3), and phycoerythrin; bioluminescent materials, such as luciferase; radioactive materials, such as 212 Bi, 14 C, 57 Co, 51 Cr, 67 Cu, 18 F, 68 Ga
  • An anti-C3 antibody as described herein may be attached to a solid support.
  • solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene.
  • immobilized anti-C3 antibodies are used in immunoassays.
  • immobilized anti-C3 antibodies are used in purification of the target antigen.
  • the disclosure encompasses a polynucleotide composition comprising a polynucleotide or polynucleotides that encode a C3 -binding agent described herein.
  • a polynucleotide or polynucleotides that encode a C3 -binding agent encompasses a polynucleotide or polynucleotides which include only coding sequences for a C3 -binding agent as well as a polynucleotide or polynucleotides which include additional coding and/or non-coding sequences.
  • the polynucleotide or polynucleotides of the disclosure can be in the form of RNA or in the form of DNA.
  • DNA includes cDNA, genomic DNA, and synthetic DNA; and can be double-stranded or single- stranded, and if single stranded can be the coding strand or non-coding (anti-sense) strand.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, provided herein is a polynucleotide or polynucleotides encoding a polypeptide or polypeptides comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:36. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:37. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:38. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:39.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:36 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:40.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:37 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:38 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:40.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:39 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:41.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:36 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:40.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:37 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:38 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:40.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:39 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:41.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NOs:77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
  • a polynucleotide or polynucleotides encoding a polypeptide or polypeptides comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149,
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:77. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 95. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:96.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 113. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:l 14. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 131. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 132.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 149. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 167. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 168.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 185. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:203. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:204.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:220. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:222. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:223.
  • the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:224. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:77 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:95 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:96.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 113 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 114. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 131 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 132.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 149 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 167 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 168.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 185 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:203 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:204.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:220 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:222 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35.
  • the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:223 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:224 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:77 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:95 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:96.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 113 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 114. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 131 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 132.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 149 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 167 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 168.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 185 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:203 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:204.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:220 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:222 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35.
  • the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:223 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:224 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35.
  • a polynucleotide comprises a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98%, or 99% identical to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. Also provided is a polynucleotide that comprises a polynucleotide that hybridizes to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, the hybridization is under conditions of high stringency as is known to those skilled in the art.
  • a polynucleotide comprises a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98%, or 99% identical to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NO s: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
  • polynucleotide that comprises a polynucleotide that hybridizes to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
  • the hybridization is under conditions of high stringency as is known to those skilled in the art.
  • a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence selected from the group consisting of: SEQ ID NOs:36-41. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:36 or SEQ ID NO:37. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:38 or SEQ ID NO:39. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:40 or SEQ ID NO:41.
  • a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence selected from the group consisting of: SEQ ID NO s: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
  • a polynucleotide comprises the coding sequence for a polypeptide fused in the same reading frame to a polynucleotide which aids in expression and secretion of a polypeptide from a host cell (e.g ., a leader sequence which functions as a secretory sequence for controlling transport of a polypeptide).
  • the polypeptide can have the leader sequence cleaved by the host cell to form a “mature” form of the polypeptide.
  • a polynucleotide comprises the coding sequence for a polypeptide fused in the same reading frame to a marker or tag sequence.
  • a marker sequence is a hexa-histidine tag (SEQ ID NO:47) (HIS- tag) that allows for efficient purification of the polypeptide fused to the marker.
  • a marker sequence is a hemagglutinin (HA) tag derived from the influenza hemagglutinin protein when a mammalian host is used.
  • the marker sequence is a FLAGTM tag.
  • a marker may be used in conjunction with other markers or tags.
  • the present disclosure also provides variants of the polynucleotides described herein, wherein the variant encodes, for example, fragments, analogs, and/or derivatives of a polypeptide.
  • the present disclosure provides a polynucleotide comprising a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98% or 99% identical to a polynucleotide sequence encoding a polypeptide described herein.
  • a polynucleotide having a nucleotide sequence at least 95% identical to a polynucleotide sequence is intended to mean that the nucleotide sequence of the polynucleotide is identical to a reference sequence except that the polynucleotide sequence can include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence.
  • a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence up to 5% of the nucleotides in the reference sequence can be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence can be inserted into the reference sequence. It is understood by those of skill in the art that an appropriate calculation would be made for other “% identical” statements, for example, 90% identical or 85% identical. These mutations of the reference sequence can occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.
  • the polynucleotide variants can contain alterations in the coding regions, noncoding regions, or both.
  • a polynucleotide variant contains alterations which produce silent substitutions, additions, or deletions, but does not alter the properties or activities of the encoded polypeptide.
  • a polynucleotide variant comprises silent substitutions that results in no change to the amino acid sequence of the polypeptide (due to the degeneracy of the genetic code).
  • a polynucleotide variant comprises one or more mutated codons comprising one or more ( e.g ., 1, 2, or 3) substitutions to the codon that change the amino acid encoded by that codon.
  • a polynucleotide variant comprises at least one silent mutation in a noncoding or a coding region of the sequence.
  • a polynucleotide variant is produced to modulate or alter expression (or expression levels) of the encoded polypeptide. In some embodiments, a polynucleotide variant is produced to increase expression of the encoded polypeptide. In some embodiments, a polynucleotide variant is produced to decrease expression of the encoded polypeptide. In some embodiments, a polynucleotide variant has increased expression of the encoded polypeptide as compared to a parental polynucleotide sequence. In some embodiments, a polynucleotide variant has decreased expression of the encoded polypeptide as compared to a parental polynucleotide sequence.
  • a polynucleotide is isolated. In some embodiments, a polynucleotide is substantially pure.
  • an expression vector comprises a polynucleotide or polynucleotides encoding a C3-binding agent described herein. In some embodiments, an expression vector or vectors comprise a polynucleotide or polynucleotides encoding a C3 -binding agent described herein. In some embodiments, an expression vector or vectors comprise a polynucleotide or polynucleotides encoding a polypeptide or polypepides that are part of a C3 -binding agent described herein.
  • a host cell comprises an expression vector or vectors comprising a polynucleotide or polynucleotides encoding a C3- binding agent described herein. In some embodiments, a host cell comprises an expression vector or vectors comprising a polynucleotide or polynucleotides encoding a polypeptide or polypeptides that are part of a C3 -binding agent described herein. In some embodiments, a host cell comprises a polynucleotide or polynucleotides encoding a C3 -binding agent described herein.
  • the present disclosure provides methods useful in a variety of applications including, but not limited to, therapeutic treatment methods, such as treatment of diseases or disorders associated with complement activation.
  • the methods comprise use of a C3 -binding agent described herein.
  • a C3 -binding agent described herein is useful in methods for inhibiting complement activation.
  • a C3-binding agent described herein is useful in methods for inhibiting C3 activity.
  • a C3-binding agent described herein is useful in methods for treating respiratory illness.
  • a C3 -binding agent described herein is useful in methods for treating severe respiratory illness.
  • a C3-binding agent described herein is useful in methods of preventing or inhibiting the development of respiratory illness. In some embodiments, a C3 -binding agent described herein is useful in methods of preventing or inhibiting the development of severe respiratory illness. In some embodiments, a C3 -binding agent described herein is useful in methods of inhibiting complement pathway activation. In some embodiments, a C3-binding agent described herein is useful in methods of treating thrombosis. In some embodiments, a C3-binding agent described herein is useful in methods of preventing or inhibiting the development of thrombosis. In some embodiments, a C3-binding agent described herein is useful in methods of treating a coronavirus infection. In some embodiments, a C3-binding agent described herein is useful in methods of treating the sequelae of a coronavirus infection.
  • a method of inhibiting complement activation in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of inhibiting complement activation of a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of inhibiting C3 activity in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein.
  • a method of inhibiting C3 activity in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • a method of treating a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3- binding agent described herein. In some embodiments, a method of treating a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of treating a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of treating a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • a method of preventing or inhibiting development of a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting development of a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting development of a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • a method of preventing or inhibiting development of a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • the severe respiratory illness is selected from the group consisting of: pneumonia, acute respiratory disease (ARD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), atypical ARDS, respiratory distress syndrome (RDS), severe acute respiratory syndrome (SARS),
  • the respiratory illness is pneumonia. In some embodiments of the methods described herein, the respiratory illness is ARD. In some embodiments of the methods described herein, the respiratory illness is ALI. In some embodiments of the methods described herein, the respiratory illness is ARDS.
  • ARDS is an acute diffuse, inflammatory lung injury, leading to increased pulmonary vascular permeability, increased lung weight, and loss of aerated lung tissue with hypoxemia and bilateral radiographic opacities, associated with increased venous admixture, increased physiological dead space and decreased lung compliance.
  • the definition identifies three mutually exclusive categories of increasingly severe ARDS based on the degree of arterial hypoxemia as measured by the PaO2/FiO2 ratio (P/F): (i) mild - P/F 201 to 300 mm Hg; (ii) moderate - P/F 101 to 200 mm Hg; and (iii) severe - P/F ⁇ 100 mm Hg.
  • the respiratory illness is atypical ARDS. Symptoms include hypoxia, shortness of breath, rapid breathing, and bluish skin coloration. For those who survive, a decreased quality of life is common. Atypical ARDS has been observed in some subjects infected with SARS-CoV-2. As used herein, “atypical ARDS” is characterized by moderate or severe hypoxia, without the feeling of shortness of breath. It is also characterized by severe hypoxia but well preserved lung gas volume, which is not seen in ARDS. In some embodiments of the methods described herein, the respiratory illness is RDS. In some embodiments of the methods described herein, the respiratory illness is SARS. In some embodiments of the methods described herein, the respiratory illness is MERS. In some embodiments of the methods described herein, the respiratory illness is COVID-19.
  • a method of treating ARDS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating ARDS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of ARDS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of ARDS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • the subject has a suspected or confirmed viral infection.
  • the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor.
  • the viral infection is caused by a coronavirus, including but not limited to, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS- CoV-1, or Middle East respiratory syndrome coronavirus (MERS-CoV), optionally wherein the virus binds human ACE2 receptor.
  • a coronavirus including but not limited to, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS- CoV-1, or Middle East respiratory syndrome coronavirus (MERS-CoV), optionally wherein the virus binds human ACE2 receptor.
  • the coronavirus is SARS-CoV-2.
  • a method of inhibiting complement pathway activation in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of inhibiting complement pathway activation in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed viral infection. In some embodiments, the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor.
  • the viral infection is caused by a coronavirus, including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor.
  • a coronavirus including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor.
  • the coronavirus is SARS-CoV-2.
  • a method of treating thrombosis in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating thrombosis in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of thrombosis in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of thrombosis in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • the thrombosis is a pulmonary embolism (PE), large blood vessel blood clot, a deep vein thrombosis (DVT), or microvascular thrombosis.
  • the thrombosis has the potential to lead to a stroke.
  • the subject has symptoms of a stroke.
  • the subject has been diagnosed with a stroke.
  • the subject has a suspected or confirmed viral infection.
  • the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor.
  • the viral infection is caused by a coronavirus, including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor.
  • a coronavirus including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor.
  • the coronavirus is SARS-CoV-2.
  • a method of treating a coronavirus infection in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of treating a coronavirus infection in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
  • the coronavirus includes but is not limited to, SARS-CoV-2, SARS- CoV-1, or MERS-CoV.
  • the coronavirus is SARS-CoV-2.
  • the infection with SARS-CoV-2 results in the patient being diagnosed with COVID-19.
  • the infection with SARS-CoV-1 results in the patient being diagnosed with SARS.
  • the infection with MERS-CoV results in the patient being diagnosed with MERS.
  • a method of treating SARS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating SARS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of SARS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of SARS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed SARS-CoV-1 infection.
  • a method of treating MERS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating MERS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of MERS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of MERS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed MERS-CoV infection.
  • a method of treating COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed SARS-CoV-2 infection.
  • the viral infection is associated with a dysregulated pro-inflammatory cytokine response, which may be referred to as “severe cytokine release syndrome” or “cytokine storm”.
  • the pro-inflammatory cytokine response includes, but is not limited to, TNF-a, IL-6, IL-10, IL-la, IL-Ib, and IL-12.
  • there is an elevated level of cytokines in a sample obtained from a subject wherein the cytokines include, but are not limited to, TNF-a, IL-6, IL-10, IL-la, IL-Ib, and IL-12.
  • the subject has lung damage, respiratory failure, kidney damage, kidney failure, liver damage, heart damage, vascular damage, thrombosis, stroke, central nervous system injury, and/or multiple organ failure.
  • the subject has one or more symptoms selected from the group consisting of: hypoxemia, cough, wheezing, dyspnea, hyperpnea, pulmonary /lung inflammation, shortness of breath, labored breathing, rapid breathing, accumulation of alveolar fluid, pulmonary edema, vascular leakage, lymphocyte infiltration in the lung, lymphopenia, fever, chills, shaking chills, increased heart rate, chest pain, low blood pressure, headache, confusion, seizures, extreme tiredness, sepsis, bluish coloring of nails or lips, toe rashes/redness, toe swelling, loss of sense of smell, loss of sense of taste, and diarrhea.
  • the subject has mild, moderate or severe hypoxemia as determined by Partial Pressure of arterial oxygen/Fraction of inspired oxygen (PaCk/FiCk) or positive end-expiratory pressure (PEEP).
  • PaCk/FiCk Partial Pressure of arterial oxygen/Fraction of inspired oxygen
  • PEEP positive end-expiratory pressure
  • the subject has severe hypoxemia with a Pa02/Fi02 of less than 100.
  • the subject is human.
  • the C3 -binding agent has at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8,) of the following properties: (a) is an antagonist of C3; (b) inhibits C3 activity; (c) inhibits C3 cleavage; (d) inhibits C3 cleavage and C3a release; (e) inhibits complement activation; (f) inhibits activation of the alternative complement pathway; (g) inhibits activation of the classical complement pathway; and (h) inhibits activation of the alternative complement pathway and classical complement pathway.
  • the C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 38G10, e.g., as exemplified in Table 1 A.
  • the C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10, e.g., as exemplified in Table IB.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(G56A), e.g., as exemplified in Table IB.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(G56T), e.g., as exemplified in Table 1C.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(N55E), e.g., as exemplified in Table 1C.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(N55Q), e.g., as exemplified in Table 1C.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 3D8, e.g., as exemplified in Table 2.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 3G8, e.g., as exemplified in Table 3.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 1502, e.g., as exemplified in Table 4.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 27A8, e.g., as exemplified in Table 5.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 28C3, e.g., as exemplified in Table 6.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 38F5, e.g., as exemplified in Table 7.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 62B11, e.g., as exemplified in Table 8.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 62F2, e.g., as exemplified in Table 9.
  • the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 63A3, e.g., as exemplified in Table 10.
  • the C3 -binding agent is an anti-C3 antibody.
  • the anti- C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25) YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO:52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising
  • the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNTGTTYNQQFTG (SEQ ID NO:48), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHEGGTTYNQQFTG (SEQ ID NO: 52), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHQGGTTYNQQFTG (SEQ ID NO:56), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
  • the anti-C3 antibody comprises: (a) a heavy chain variable region of SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, or SEQ ID NO:224; and (b) a light chain variable region of SEQ ID NO:35.
  • the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:33 and a light chain variable region of SEQ ID NO:35.
  • the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:34 and a light chain variable region of SEQ ID NO:35.
  • the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:222 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:223 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:224 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10. In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(G56A).
  • the anti-C3 antibody is Hz38G10(G56T). In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(N55E). In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(N55Q).
  • the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:37. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:39. In some embodiments of the methods described herein, the anti-C3 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:37 and a light chain comprising the amino acid sequence of SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:39 and a light chain comprising the amino acid sequence of SEQ ID NO:41.
  • the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:37. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:39. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:37 and a polypeptide of amino acids set forth in SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO: 39 and a polypeptide of amino acids set forth in SEQ ID NO:41.
  • a method comprises using a C3-binding agent in combination therapy.
  • combination therapy includes, but is not limited to, (i) combination with a medical device, for example, a ventilator or ECMO machine and (ii) combination with at least one additional therapeutic agent.
  • combination therapy comprises administration of a C3-binding agent in combination with at least one additional therapeutic agent and the usage of a medical device.
  • a method comprises administering a C3-binding agent described herein in combination with usage of a medical device.
  • a method comprises administering a C3-binding agent described herein in combination with at least one additional therapeutic agent.
  • Combination therapy using agents with different mechanisms of action may result in additive or synergetic effects.
  • Combination therapy may allow for a lower dose of each agent than is used in monotherapy, thereby reducing toxic side effects and/or increasing the therapeutic index of the agent(s).
  • Combination therapy may decrease the likelihood that resistance to an agent will develop.
  • the combination of a C3 -binding agent described herein and at least one additional therapeutic agent results in additive or synergistic results.
  • the combination therapy results in an increase in the therapeutic index of the C3 -binding agent.
  • the combination therapy results in an increase in the therapeutic index of the additional therapeutic agent(s).
  • the combination therapy results in a decrease in the toxicity and/or side effects of the C3 -binding agent.
  • the combination therapy results in a decrease in the toxicity and/or side effects of the additional therapeutic agent(s).
  • a combination treatment comprises one additional therapeutic agent or two or more additional therapeutic agents.
  • treatment with a C3-binding agent can occur prior to, concurrently with, or subsequent to administration of the additional therapeutic agents.
  • combined administration includes co-administration, either in a single pharmaceutical formulation or using separate formulations, or consecutive administration in either order but generally within a time period such that all active agents can exert their biological activities.
  • preparation of agents and/or dosing schedules for additional therapeutic agents are according to manufacturers' instructions or as determined empirically by the skilled practitioner.
  • a C3-binding agent is administered to a subject in combination with one or more additional therapeutic agents.
  • an additional therapeutic agent is compstatin or an analog or derivative of compstatin (e.g ., POT-4; APL-2, AMY- 101).
  • an additional therapeutic agent is a C5 inhibitor.
  • a C5 inhibitor is selected from the group including, but not limited to, eculizumab (SOLIRIS), LFG316, or Zimura (anti-C5 aptamer).
  • an additional therapeutic agent is a properdin inhibitor (e.g., an anti-properdin antibody).
  • an additional therapeutic agent is a Factor D inhibitor.
  • a Factor D inhibitor is an anti-Factor D antibody (e.g, lampalizumab).
  • an additional therapeutic agent is an anti-viral agent.
  • the anti-viral agent is remdisivir.
  • the additional therapeutic agent is one or more of: dexamethasone, remdesivir, baricitinib in combination with remdesivir, favipiravir, merimepodib, an anticoagulation drug selected from low-dose heparin or enoxaparin, bamlanivimab, a combination of bamlanivimab and etesevimab, a combination of casirivimab and imdevimab, convalescent plasma, an mRNA SARS-CoV-2 vaccine (e.g., those produced by Moderna and Pfizer), an attenuated SARS-CoV-2 virus vaccine, a dead SARS-CoV-2 virus vaccine, a viral vaccine against SARS-CoV-2 (e.g., an adenoviral vaccine such as those produced by Johnson & Johnson and Astrazeneca), an antibody or fragment thereof or small molecule that blocks interaction between hACE2 and the spike protein of SARS-CoV-2, protea
  • a C3 -binding agent described herein and at least one additional therapeutic agent may be administered in any order or concurrently.
  • a C3-binding agent is administered to subjects that have previously undergone treatment with a therapeutic agent.
  • a C3-binding agent and a second therapeutic agent are administered substantially simultaneously or concurrently.
  • the appropriate dosage of a C3-binding agent of the present disclosure depends on the disorder or disease to be treated, the severity and course of the disorder or disease, the responsiveness of the disorder or disease, whether the agent is administered for therapeutic or preventative purposes, previous therapy, the subject’s condition, the subject's clinical history, and so on.
  • a C3-binding agent can be administered one time or over a series of treatments lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved.
  • the dose of a C3 -binding agent depends on the site of administration.
  • the dose of a C3-binding agent depends on the delivery system.
  • compositions comprising a C3-binding agent described herein.
  • present disclosure also provides pharmaceutical compositions comprising a C3-binding agent described herein and a pharmaceutically acceptable vehicle.
  • a pharmaceutical composition comprises an anti-C3 antibody described herein and a pharmaceutically acceptable vehicle.
  • a pharmaceutical composition comprises antibody Hz38G10 and a pharmaceutically acceptable vehicle.
  • a pharmaceutical composition comprises antibody Hz38G10(G56A) and a pharmaceutically acceptable vehicle.
  • Formulations are prepared for storage and use by combining a purified antibody or agent of the present disclosure with a pharmaceutically acceptable vehicle (i e.g ., a carrier or excipient).
  • a pharmaceutically acceptable vehicle i e.g ., a carrier or excipient.
  • pharmaceutically acceptable carriers, excipients, and/or stabilizers to be inactive ingredients of a formulation or pharmaceutical composition.
  • Suitable pharmaceutically acceptable vehicles include, but are not limited to, nontoxic buffers such as phosphate, citrate, and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives such as octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens, such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3- pentanol, and m-cresol; low molecular weight polypeptides ( e.g ., less than about 10 amino acid residues); proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine
  • the formulation is in the form of an aqueous solution. In some embodiments, the formulation is stored in a lyophilized or in an alternative dried form.
  • the binding agents of the present disclosure can be formulated in any suitable form for delivery to a target cell/tissue.
  • a C3-binding agent can be formulated as a liposome, microparticle, microcapsule, albumin microsphere, microemulsion, nano-particle, nanocapsule, or macroemulsion.
  • the pharmaceutical formulation includes an agent of the present disclosure complexed with liposomes.
  • liposomes are known to those of skill in the art. For example, some liposomes can be generated by reverse phase evaporation with a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE).
  • PEG-PE PEG-derivatized phosphatidylethanolamine
  • a C3-binding agent is formulated as a sustained-release preparation.
  • sustained-release preparations include semi- permeable matrices of solid hydrophobic polymers containing an agent, where the matrices are in the form of shaped articles (e.g., films or microcapsules).
  • Sustained- release matrices include but are not limited to polyesters, hydrogels such as poly(2- hydroxy ethyl -methacrylate) or poly(vinyl alcohol), polylactides, copolymers ofL- glutamic acid and 7 ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOTTM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.
  • polyesters such as poly(2- hydroxy ethyl -methacrylate) or poly(vinyl alcohol), polylactides, copolymers ofL- glutamic acid and 7 ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, de
  • compositions or formulations of the present disclosure can be administered in any number of ways for either local or systemic treatment. Administration can be (i) topical by epidermal or transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders, (ii) pulmonary by inhalation or insufflation of powders or aerosols, including by nebulizer, intratracheal, and intranasal, (iii) oral, or (iv) parenteral including intravenous, intraarterial, intratumoral, subcutaneous, intraperitoneal, intramuscular (e.g, injection or infusion), or intracranial (e.g, intrathecal or intraventricular).
  • parenteral including intravenous, intraarterial, intratumoral, subcutaneous, intraperitoneal, intramuscular (e.g, injection or infusion), or intracranial (e.g, intrathecal or intraventricular).
  • Anti-C3 antibodies were generated using human C3 as the immunogen.
  • Lymphocytes Single cell suspensions of lymphocytes were obtained from the spleens and lymph nodes of immunized mice after the individual animals had been determined to have suitable antibody titers. Lymphocytes were fused with murine myeloma cells by standard methods. Cells were dispersed into 96-well plates in HAT-containing selection media. Cell supernatants (approximately 10,000) were screened by ELISA for binding to human C3 and cynomolgus monkey (“cyno”) C3. Approximately 1600 cell clones were selected based on binding assay results.
  • C3a is produced by the cleavage of C3 by the C3 convertase complex.
  • the C3a assay was based upon a previously described method (Loyet et al., 2014, J Pharmacol and Exp Ther ⁇ 351:527-537). Briefly, 20 pL of solution of 50 nM human C3 (Complement Technologies) in GVB (bovine skin gelatin in veronal buffer) was mixed with 20 pL test anti-C3 antibodies or controls diluted in GVB for 10 minutes.
  • Controls included (i) no C3; (ii) no Factor B and Factor D; (iii) no inhibitor; and (iv) compstatin as positive control.
  • 100 nM human Factor B and human 100 nM Factor D (Complement Technologies) in 0.05 M EGTA/0.1 M MgC1 2 /GVB were added to the C3/Ab samples and the mixtures were incubated for an additional 10 minutes.
  • 20 pL of 50% rabbit serum (diluted 1 :2 in GVB) was added and incubated for 10 minutes, followed by addition of 20 pL of 0.5 M EDTA to quench the reaction.
  • C3a des Arg The addition of rabbit serum cleaves C3a produced by C3 convertase in the reaction mixtures to C3a des Arg (C3a des Arg has been found to be more stable than C3a).
  • C3 des Arg concentrations were subsequently measured by ELISA. Briefly, a mouse anti-C3a/C3a des Arg antibody (Therm oFisher) was coated on ELISA plates at 1.0 pg/mL and incubated overnight at 4°C. The following day, the plates were washed and 60 pL of SuperBlock blocking buffer (Pierce) was added to the plates and incubated for 60 minutes at room temperature. Prior to adding samples, blocking buffer was decanted from the wells.
  • SuperBlock blocking buffer Pieris
  • test anti-C3 antibodies were ranked for their ability to inhibit C3a des Arg production.
  • exemplary antibodies were observed to strongly inhibit C3 cleavage and C3a release.
  • the activities of three exemplary antibodies, 38G10, Ab2 (3D8), and Ab3 (15C12), at concentrations ranging from 0.1 to 1000 nM, are shown in Figure 1.
  • Example 3
  • Hemolytic assays have traditionally been used to assess the functional activity of the complement system.
  • AP alternative pathway
  • rabbit erythrocytes were included with C3-depleted human serum and 50 nM of human C3 in the presence of increasing concentrations of anti-C3 antibodies 38G10, Ab2, and Ab3.
  • CP classical pathway
  • IgM-coated sheep erythrocytes were incubated with C3- depleted human serum and increasing concentration of anti-C3 antibodies 38G10, Ab2, and Ab3 in the presence of 10 nM human C3.
  • Hemolysis was determined by reading the absorbance of the supernatant at 412 nm. Results were expressed as a percentage of hemolysis observed in control samples without any inhibitor and can be calculated into IC50s.
  • Results of the AP and CP assays using human C3 and cyno C3 are shown in Figure 2 for antibodies 38G10, Ab2 (3D8), and Ab3 (1502) (% hemolysis). The results are also summarized in Table 11.
  • binding affinities of exemplary anti-C3 antibodies to human C3 and cyno C3 were measured using a Biacore system (GE Healthcare LifeSciences). Briefly, anti- Fc antibody (Sigma-Aldrich) was immobilized on all four flow cells of a CM5 chip using amine coupling reagents (GE Healthcare LifeSciences). Antibodies were captured on flow cells 2, 3, and 4 using flow cell 1 as a reference. Concentrations ranging from 0.62 - 40 nM of human C3 or cyno C3 were injected at a flow rate of 50 ⁇ L/min at 37°C. Kinetic data were collected over time and fit using the simultaneous global fit equation to yield affinity constants (KD values) for each antibody.
  • KD values affinity constants
  • Antibody 38G10 bound to human C3 with a KD of 1.9 x 10 '10 M and to cyno C3 with a KD of 1.6 X lO -10 M.
  • Table 12 shows the binding affinities for antibodies 38G10, 3D8, and 15C12.
  • the anti-C3 antibodies described herein were characterized for their ability to bind a number of other components of the complement cascade. Antibodies were screened for binding to C3, C3a, C3b, C3c, C3d, iC3b, and Factor Bb. A representative set of the results is shown in Table 13. All the anti-C3 antibodies included in Table 13 were able to block complement activation. The data are relative to the binding of the negative control mIgG2 antibody.
  • Antibody 38G10 was humanized by methods known to those skilled in the art and humanized 38G10 is referred to herein as Hz38G10.
  • Antibody 38G10 was found to have a potential glycosylation site in CDR2 of the heavy chain variable region, YIYPHNGGTTYNONFTG (SEQ ID NO: 10).
  • the consensus glycosylation site for N- linked glycans is N-X-S/T, wherein X can be any amino acid except proline.
  • the heavy chain CDR2 was reengineered to remove the glycosylation site resulting in antibody Hz38G10 comprising a heavy chain CDR2 comprising YIYPHNGGTTYNQQFTG (SEQ ID NO:25).
  • the heavy chain variable sequence of antibody 38G10 is SEQ ID NO:31 and the light chain variable sequence is SEQ ID NO:32; the heavy chain variable sequence of antibody Hz38G10 is SEQ ID NO:33 and the light chain variable sequence is SEQ ID NO:35; CDRs for 38G10 and Hz38G10 are disclosed in Tables 1A and IB.
  • the binding affinity of Hz38G10 was determined using a Biacore system as described herein and compared with the binding affinity of the parental 38G10 antibody.
  • Antibody Hz38G10 had a binding affinity to human C3 of 3 x 10 '10 M and to cyno C3 of 1.8 x 10 '9 M as compared to the parental antibody’s binding affinity of approximately 1 x 10 '10 M to both human and cyno C3 (at 37°C).
  • a Biacore system was used to assess the binding affinities of the different molecules to human C3. Binding affinity of Hz38G10 to human C3 at 25°C and 37°C was carried out as described in Example 4. Evaluation of COMP or COMP -PEG binding to human C3 was carried out as follows. Human C3 was immobilized on flow cell 2 of a CM5 chip using amine-coupling reagents (GE Healthcare LifeSciences) and flow cell 1 (immobilized with a similar amount of human IgG) was used as a reference. Concentrations ranging from 1.56 - lOOnM of COMP were injected at a flow rate of 50 pL/min for evaluation of binding affinity at 25°C and 37°C.
  • COMP-PEG concentration range of 25 - 400 nM
  • COMP -PEG binding to human C3 was also evaluated at 25°C with modifications.
  • rat anti-PEG antibody Abeam Inc.
  • Human C3 concentration range 12 - 100 nM
  • Kinetic data were collected over time and fit using the simultaneous global fit equation to yield affinity constants (KD values) for each antibody.
  • Hz38G10 was assessed in alternative and classical pathway-dependent hemolysis assays as described herein and compared to COMP and COMP -PEG. Hz38G10 and COMP or antibody Hz38G10 and COMP -PEG were tested for their respective abilities to inhibit hemolysis in the assays.
  • Hz38G10 variants were generated with amino acid substitutions within the heavy chain CDR2 at position N55 or G56 of SEQ ID NO:33.
  • the Hz38G10 variants contained (i) a N55E mutation; (ii) a N55Q mutation; (iii) a G56A mutation; or (iv) a G56T mutation.
  • the heavy chain variable region sequence of antibody Hz38G10(G56A) is SEQ ID NO:34
  • Hz38G10(G56T) is SEQ ID NO:222
  • Hz38G10(N55E) is SEQ ID NO:223
  • Hz38G10(N55Q) is SEQ ID NO:224
  • the light chain variable region sequence for all variants is SEQ ID NO:35
  • CDRs are disclosed in Tables 1B-1C.
  • scFv molecules based on the heavy chain variable region (VH) and the light chain variable region (VL) of Hz38G10 were designed in two orientations and with or without disulfide stabilization.
  • the four molecules were (i) VL-linker-VH scFv, (ii) VH-linker- VL scFv, (iii) VL-linker-VH dsscFv; and (iv) VH-linker-VL dsscFv, wherein dsscFv indicates a scFv with an engineered disulfide bond between the VL and VH regions.
  • VL-linker-VH dsscFv The sequence of Hz38G10 dsscFv is disclosed herein as SEQ ID NO:228.
  • Hz38G10 dsscFv antibodies were produced via expression as inclusion bodies.
  • BL21(DE3) E. coli were transformed with a plasmid encoding Hz38G10 VL-linker-VH dsscFv, referred to hereafter as Hz38G10 dsscFv.
  • Transformed BL21(DE3) E. coli were harvested by low speed centrifugation. The bacterial pellet was resuspended by magnetic stirring in 50 mM Tris-HCl pH 7.5 for 15 minutes at room temperature. For cell disruption, the suspension was passed 3 times through a microfluidizer at 200 Bar.
  • Inclusion bodies were recovered from broken cells by centrifugation at 4°C in a fixed angle rotor operating at 13,000 xg for 45 minutes. The resulting inclusion body pellet was washed with cold Milli-Q® water until the lipid and cell wall layers were removed. The washed inclusion body pellet was resuspended in 5 volumes of Milli-Q® water by magnetic stirring and collected by centrifugation at 13,000 xg for 30 minutes. Washed inclusion bodies were solubilized in 10 volumes of 8 M guanidine-HCl, 50 mM Tris- HCl pH 7.5, 1 mM TCEP using magnetic stirring and incubated overnight at 4°C.
  • Solubilized inclusion bodies were diluted dropwise 1 :25 into refold buffer (3 M urea, 50 mM Tris-HCl pH 8.0, 160 mM L- arginine, 1 mM cysteine, 3 mM cystamine) under gentle magnetic stirring at room temperature to a final concentration of 0.1 mg/ml. Refolding was carried out for 48 hours at 4°C and disulfide bond formation was monitored by SDS-PAGE under nonreducing conditions.
  • refold buffer 3 M urea, 50 mM Tris-HCl pH 8.0, 160 mM L- arginine, 1 mM cysteine, 3 mM cystamine
  • the solution containing the refolded dsscFv was concentrated 5- fold by ultrafiltration/diafiltration (UF/DF) on a 0.22 m 2 10,000 DaNMWCO tangential flow filtration membrane.
  • the concentrate was cleared of misfolded dsscFv by rapid acidification to pH 2.8 with 4 volumes of 100 mM glycine pH 2.5.
  • the solution was held at pH 2.8 as the solution was further concentrated 5-fold. After 15 minutes, the pH was quickly returned to neutral with addition of concentrated Tris-HCl solution.
  • the resulting acid-treated solution was buffer-exchanged with diavolumes of Protein-L affinity binding buffer (50 mM Tris-HCl pH 7.5, 50 mM NaCl). Recovered diafilitration product was cleared of precipitate by centrifugation at 13,000 xg for 45 minutes at 4°C and subsequently filtered using a 0.2 pm membrane.
  • Hz38G10 dsscFv antibodies were purified in two chromatography steps: (i) a Protein L affinity column for capture and (ii) a cation exchange (CEX) column for polishing.
  • a Protein L affinity column for capture and (ii) a cation exchange (CEX) column for polishing.
  • CEX cation exchange
  • the suspension containing Hz38G10 dsscFv was applied to a 5 mL MiniChrom TOYOPEARL® AF-rProtein L-650F column (Tosoh Bioscience), washed extensively with Tris-buffered saline, and eluted with a step gradient of 100 mM glycine pH 2.5.
  • elution pool (Pool M) was dialyzed against CEX binding buffer (20 mM sodium acetate pH 5.0). Dialyzed Pool M was applied to a HiTrap® SP HP cation exchange column (GE Healthcare), washed with 20 mM sodium acetate, pH 5.0, and eluted with an isocratic gradient of 50 sodium chloride in 20 mM sodium acetate, pH 5.0.
  • Hz38G10 dsscFv-enriched fractions were pooled and evaluated by SDS-PAGE and analytical SEC methods. Subsequently, a Hz38G10(G56A) dsscFv antibody (SEQ ID NO:229) was generated following the same general methods.
  • the solubility parameters of Hz38G10 dsscFv were investigated.
  • the purified antibodies were dialyzed into buffer consisting of 10 mM sodium phosphate, 40 mM sodium chloride, 0.03% polysorbate 20, and 5% sucrose, pH 6.2.
  • the solution was micro-concentrated using 10,000 Da NMWCO spin concentrator at 3,000 xg at room temperature. Protein concentration was measured on an Agilent Cary 60 UV-VIS Solo VPE spectrophotometer. Protein concentration reached 229 mg/ml with no visible aggregation.
  • Hz38G10 dsscFv were compared to Hz38G10 intact antibody and a Fab version of Hz38G10 in several assays. Binding affinity to human C3 was assessed using a Biacore system as described herein. In addition, inhibition of the alternative pathway and classical pathway by the three antibodies was assessed in hemolysis assays as described herein. [00289] The results are summarized in Table 18.
  • Hz38G10 dsscFv had a similar KD as compared to the intact antibody.
  • Hz38G10 dsscFv had a similar level of inhibition of hemolytic activity in both the AP and CP pathways as compared to the intact antibody or the Fab version.
  • the intact Hz38G10 antibody has two antigen-binding sites, while the Fab and dsscFv antibodies have only one antigenbinding site.
  • 3D8 Heavy chain variable region (SEQ ID NO:77) EIQLQQSGAELVKPGASVKISCKASGYSFTGYNMNWVKQSHGKSLEWIGNINPYYGST NYNQKFKGKATLTVDKSSTTAYMQLDSLTSEDSAVYYCARGYYGGNYPFAYWGQGTLV TVSA
  • 3G8 Heavy chain variable region (SEQ ID NO:95) EIQLQQSGAELVKPGASVKISCMASGYSFTGYNMNWVKQSHGKGLEWIGNINPYYDST SYNQKFKGKATLTVDKSSSTAYMQLNSLTSEDSAVYYCARENYDFVGFAYWGQGTLVT VSA
  • 62F2 Heavy chain variable region (SEQ ID NO:203) DVQLQESGPDLVKPSQSLSLTCTVTGYS ITSGYSLHWIRQFPGNKLEWMGYIHYSGST NYNPSLKSRISITRDTSKNQFFLKLNSVTSEDTATYHCARAWDYLDYWGQGTTLTVSS

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Abstract

The present disclosure generally relates to methods of treating, preventing, or inhibiting the development of respiratory illness (e.g., a severe respiratory illness) and similar respiratory diseases. In addition, the disclosure relates to treating, preventing, or inhibiting the development thrombosis and resulting sequelae such as stroke. The methods comprise administering agents that bind complement component C3 (C3), particularly antibodies that bind human C3.

Description

THERAPEUTIC USES OF C3-BINDING AGENTS
CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims the priority benefit of U.S. Provisional Application No. 63/022,121, filed May 8, 2020, which is hereby incorporated by reference herein in its entirety.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on May 5, 2021, is named 47702-0103W01_SL.txt and is 203,883 bytes in size.
FIELD OF THE INVENTION
[0003] The present disclosure generally relates to methods of treating, preventing, or inhibiting the development of respiratory illness and diseases. In addition, the disclosure relates to treating, preventing, or inhibiting the development of thrombosis and resulting sequelae such as stroke. The methods comprise administering agents that bind complement component C3 (C3), particularly antibodies that bind human C3.
BACKGROUND
[0004] The complement system functions as an immune surveillance system and is part of the innate immune response that rapidly responds to bacterial and/or viral infections. Activation of the complement system promotes inflammation, both acute and chronic, and often results in elimination of pathogens. The complement system includes more than 30 cell-associated and circulating proteins ( e.g ., Cl, Clq, Clr, Cls, C2, C3, C3a, C3b, C4, C5, Factor B, Factor D, Factor H, Factor I). There are three main pathways that activate complement, the classical pathway (CP), the lectin pathway (MBL), and the alternative pathway (AP).
[0005] The three complement pathways are initiated by different factors, each resulting in the cleavage of complement component C3. The classical complement pathway is activated when the Cl complex binds specific antigen-antibody complexes, often immunoglobulin M (IgM), IgG3, or IgGl. This induces a conformational change in the Cl complex, allowing it to cleave C4 and C2 to generate the C4bC2b complex. C4bC2b acts as the C3 convertase of the classical pathway. The lectin complement pathway is activated when mannose-binding lectin (MBL) binds mannose-containing polysaccharides on microorganisms, initiating the cleavage of C4 and C2 by the MBL- MBL-associated serine protease complex. As in the classical pathway, the C4bC2b complex forms the C3 convertase of the lectin pathway. The oldest evolutionary signaling pathway of the three, the alternative complement pathway acts both independently of, and as an amplification loop for, the classical and lectin pathways.
The alternative complement pathway undergoes low-level self-activation through the slow, spontaneous hydrolysis of C3. Once hydrolyzed, C3(H20) binds complement Factor B, which is subsequently cleaved by complement Factor D into C3(H20)Bb, forming the C3 convertase of the alternative complement pathway. These different pathways converge with the cleavage of complement component C3 into C3a and C3b. C3b participates in the formation of the C5 convertase which, in turn, cleaves C5 to C5a and C5b. C3a and C5a, termed anaphylatoxins, are pleiotropic inflammatory mediators. C5b initiates the formation of the terminal C5b-9 complement complex, called the membrane attack complex (MAC). MAC can trigger cell death via phagocytosis, inflammation, and cell lysis. (. Immunobiology : The Immune System in Health and Disease. 5th edition; Chapter: The complement system and innate immunity ; New York: Garland Science; 2001.)
[0006] The coagulation system functions to mediate blood clotting. Similar to the immune system, it is a highly complex system made up of a large number of blood cells and soluble factors/proteins. Normal coagulation system homeostasis represents a balance between (i) the pro-coagulant processes that are responsible for clot formation, (ii) mechanisms that inhibit the clotting process beyond an injury site, and (iii) fibrolysis, which involves a distinct enzymatic cascade that lead to removal of clots. Imbalance of the coagulation system and/or fibrolysis may occur during critical illness, in post-operative situations, or in some diseases. Dysfunction of the coagulation system is a major contributor to thrombosis - the pathological formation of thrombi in blood vessels.
[0007] Cross-talk and interactions between the complement system and the coagulation system have been observed and further research is on-going. See, e.g ., Markiewski et ah, 2007, Trends in Immunology, 28:184-192; Oikonomopoulou, etal. , 2012, Seminars in Immunopathol., 34:151-165. However, dysregulation of one or both of these pathways, or the components thereof, for example, during a viral infection, can result in severe clinical manifestations especially in the context of diseases with inflammatory pathogenesis.
[0008] There exists an urgent need for improved methods for treating diseases associated with inflammatory pathogenesis, such as severe respiratory illnesses and the development of thrombosis and resulting sequelae such as stroke.
BRIEF SUMMARY
[0009] A pro-inflammatory response is often observed as part of the normal immune response to a viral infection. The pro-inflammatory response includes the production of cytokines and chemokines, which in turn recruit additional immune cells including leukocytes, macrophages, monocytes, and dendritic cells. The subsequent effector functions of these effector cells and soluble cellular factors contribute to the first line of defense against the virus. Generally, these events contribute to the reduction of viral replication and to the limitation of viral spread. However, in some cases the viral infection induces an overly aggressive pro-inflammatory response, which in combination with an insufficient balancing of an anti-inflammatory response, can lead to a series of events called a “cytokine storm”. Symptoms of a cytokine storm include, but are not limited to, fever, fatigue, loss of appetite, muscle and joint pain, nausea, vomiting, diarrhea, rashes, fast breathing, rapid heartbeat, low blood pressure, seizures, headache, confusion, delirium, hallucinations, tremor, and loss of coordination. A cytokine storm is often associated with a surge of activated immune cells into the lungs and over-production of cytokines/chemokines. The resulting lung inflammation and fluid buildup can lead to respiratory distress, lung damage, a variety of acute respiratory diseases, and even death.
[0010] Several respiratory viruses, particularly some strains of coronaviruses, some strains of influenza viruses, and some strains of respiratory syncytial viruses have been documented to be associated with cytokine storms and/or acute respiratory illnesses. Influenza viruses include, but are not limited to, H5N1 and H1N1 2009. Coronaviruses include, but are not limited to, SARS-CoV-1 (severe acute respiratory syndrome coronavirus 1), SARS-CoV-2, and MERS-CoV (Middle East respiratory syndrome coronavirus). Infection with a coronavirus may lead to severe disease and the syndromes are generally referred to as SARS (SARS-CoV-1), COVID-19 (SARS-CoV- 2), and MERS (MERS-CoV).
[0011] In one aspect, the present disclosure provides methods of treating, preventing, or inhibiting the development of respiratory illnesses and diseases. In some embodiments, the methods relate to treating, preventing, or inhibiting the development of severe respiratory illnesses and diseases. In addition, the disclosure provides methods of treating, preventing, or inhibiting the development of thrombosis and resulting sequelae such as stroke. In some embodiments, a respiratory illness is associated with elevated levels of pro-inflammatory cytokines and/or chemokines. In some embodiments, a severe respiratory illness is associated with elevated levels of pro- inflammatory cytokines and/or chemokines. In some embodiments, thrombosis is associated with elevated levels of pro-inflammatory cytokines and/or chemokines. In some embodiments, a respiratory illness is associated, either directly or indirectly, with dysregulation of the complement pathway. In some embodiments, a severe respiratory illness is associated, either directly or indirectly, with dysregulation of the complement pathway. In some embodiments, thrombosis is associated, either directly or indirectly, with dysregulation of the complement pathway. In some embodiments, the dysregulation of the complement pathway is associated with elevated levels of pro- inflammatory cytokines. In some embodiments, the respiratory illness is associated with a suspected or confirmed viral infection. In some embodiments, the severe respiratory illness is associated with a suspected or confirmed viral infection. In some embodiments, the thrombosis or sequelae thereof is associated with a suspected or confirmed viral infection. In some embodiments, the viral infection is caused by an influenza virus, a coronavirus, or a respiratory syncytial virus. In some embodiments, the suspected or confirmed viral infection is a coronavirus infection. In some embodiments, the suspected or confirmed viral infection is a SARS-CoV-2 infection. In some embodiments, a respiratory illness is associated with a suspected or confirmed coronavirus infection. In some embodiments, a severe respiratory illness is associated with a suspected or confirmed coronavirus infection. In some embodiments, a respiratory illness is associated with a suspected or confirmed SARS-CoV-2 infection.
In some embodiments, a severe respiratory illness is associated with a suspected or confirmed SARS-CoV-2 infection. In some embodiments, thrombosis or sequelae thereof is associated with a suspected or confirmed coronavirus infection. In some embodiments, thrombosis or sequelae thereof is associated with a suspected or confirmed SARS-CoV-2 infection.
[0012] In some embodiments of the methods described herein, a method of treating a respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of an agent that specifically binds complement component C3 (referred to herein as a “C3 -binding agent”). In some embodiments of the methods described herein, a method of treating a severe respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of an agent that specifically binds complement component C3. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of a respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of a severe respiratory illness in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments, the severe respiratory illness is selected from the group consisting of: pneumonia, acute respiratory disease (ARD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), atypical ARDS, respiratory distress syndrome (RDS), severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), and coronavirus 2019 (COVID-19). In some embodiments of the methods described herein, a method of treating ARDS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of ARDS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments, the subject has a suspected or confirmed viral infection.
[0013] In some embodiments of the methods described herein, a method of inhibiting complement pathway activation in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of inhibiting complement pathway activation in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent, wherein the subject has a suspected or confirmed viral infection.
[0014] In some embodiments of the methods described herein, a method of treating thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of treating thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent, wherein the subject has a suspected or confirmed viral infection. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of thrombosis in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent, wherein the subject has a suspected or confirmed viral infection. In some embodiments, the thrombosis is a pulmonary embolism (PE), large blood vessel blood clot, a deep vein thrombosis (DVT), or microvascular thrombosis. In some embodiments, the thrombosis has the potential to lead to a stroke. In some embodiments, the subject has symptoms of a stroke or has been diagnosed with a stroke.
[0015] In some embodiments of the methods described herein, the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus. In some embodiments, the viral infection is caused by a coronavirus. In some embodiments, the coronavirus is selected from the group consisting of: SARS-CoV-2, SARS-CoV-1, or MERS-CoV. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the viral infection is caused by SARS-CoV-2. In some embodiments, the subject has been diagnosed with the syndrome/disease that is caused by SARS-CoV-2 - COVID-19. [0016] In some embodiments of the methods described herein, a method of treating a coronavirus infection in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of treating a coronavirus infection or the resulting complications and/or sequelae of a coronavirus infection in a human subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent. In some embodiments, the coronavirus is selected from the group consisting of: SARS- CoV-2, SARS-CoV-1, or MERS-CoV. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, an infection with SARS-CoV-2 results in COVID-19.
[0017] In some embodiments of the methods described herein, a virus infection is associated with a dysregulated pro-inflammatory cytokine response or cytokine storm. In some embodiments, a coronavirus infection is associated with a dysregulated pro- inflammatory cytokine response or cytokine storm.
[0018] In some embodiments of the methods described herein, a method of treating SARS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of SARS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments, the subject has a suspected or confirmed SARS-CoV-1 infection.
[0019] In some embodiments of the methods described herein, a method of treating MERS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of MERS in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments, the subject has a suspected or confirmed MERS-CoV infection.
[0020] In some embodiments of the methods described herein, a method of treating COVID-19 in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments of the methods described herein, a method of preventing or inhibiting the development of COVID-19 in a human subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent. In some embodiments, the subject has a suspected or confirmed SARS-CoV-2 infection.
[0021] In some embodiments of the methods described herein, the subject has lung damage, respiratory failure, kidney damage, kidney failure, liver damage, heart damage, vascular damage, thrombosis, stroke, central nervous system injury, and/or multiple organ failure. In some embodiments of the methods described herein, the subject has one or more symptoms selected from the group consisting of: hypoxemia, cough, wheezing, dyspnea, hyperpnea, pulmonary /lung inflammation, shortness of breath, labored breathing, rapid breathing, accumulation of alveolar fluid, pulmonary edema, vascular leakage, lymphocyte infiltration in the lung, lymphopenia, fever, chills, shaking chills, increased heart rate, chest pain, low blood pressure, headache, confusion, seizures, extreme tiredness, sepsis, bluish coloring of nails or lips, toe rashes/redness, toe swelling, loss of sense of smell, loss of sense of taste, and diarrhea. In some embodiments of the methods described herein, the subject has mild, moderate or severe hypoxemia as determined by Partial Pressure of arterial oxygen/Fraction of inspired oxygen (PaC2/FiC2) or positive end-expiratory pressure (PEEP). In some embodiments of the methods described herein, the subject has severe hypoxemia with a PaCh/FiCh of less than 100.
[0022] In some embodiments of the methods described herein, a method comprises administering the C3 -binding agent as part of a combination therapy. In some embodiments, the combination therapy includes the use of a medical device. In some embodiments, the medical device is a ventilator or an ECMO machine. In some embodiments, the combination therapy includes the use of at least one additional therapeutic agent. In some instances, the additional therapeutic agent is one or more of: dexamethasone, remdesivir, baricitinib in combination with remdesivir, favipiravir, merimepodib, an anticoagulation drug selected from low-dose heparin or enoxaparin, bamlanivimab, a combination of bamlanivimab and etesevimab, a combination of casirivimab and imdevimab, convalescent plasma, an mRNA SARS-CoV-2 vaccine (e.g., those produced by Moderna and Pfizer), an attenuated SARS-CoV-2 virus vaccine, a dead SARS-CoV-2 virus vaccine, a viral vaccine against SARS-CoV-2 (e.g., an adenoviral vaccine such as those produced by Johnson & Johnson and Astrazeneca), an antibody or fragment thereof or small molecule that blocks interaction between hACE2 and the spike protein of SARS-CoV-2, protease inhibitors targeting SARS- CoV-2 S cleavage sites, viral fusion inhibitors, EK1C4, nelfmavir mesylate, furin inhibitors, or any other agent described in Huang et al., Acta Pharmacol ogica Sinica (2020) 41:1141-1149; https://doi.org/10.1038/s41401-020-0485-4), or unmethylated CpG dinucleotides in combination with any of these agents.
[0023] In some embodiments of the methods described herein, a method comprises administering to the subject a C3-binding agent described herein and at least one additional therapeutic agent.
[0024] In some embodiments of methods described herein, as well as other aspects and embodiments described herein, the subject is human.
[0025] In another aspect, the present disclosure provides one or more agents that bind C3, referred to herein as “a C3-binding agent” or “C3-binding agents.” In some embodiments, the C3-binding agents bind human C3 (e.g., SEQ ID NO:l or SEQ ID NO:2). In some embodiments, the C3-binding agents bind cynomolgus monkey (“cyno”) C3 (e.g., SEQ ID NO:7 or SEQ ID NO:8). In some embodiments, the C3- binding agents bind human C3 and cyno C3. In some embodiments, the C3-binding agents comprise an antibody. In some embodiments, the C3-binding agents comprise an antibody that binds human C3. In some embodiments, the C3-binding agents comprise an antibody that binds cyno C3. In some embodiments, the C3-binding agents comprise an antibody that binds human C3 and cyno C3. In some embodiments, the C3-binding agents comprise an antibody that binds human C3 and does not bind human C3b. In some embodiments, the C3-binding agents do not bind human C3b at a detectable level.
[0026] In some embodiments, the C3 -binding agents bind human C3 and have at least one or more (e.g, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) of the following properties: (a) binds to cyno C3, (b) is an antagonist of C3, (c) inhibits C3 activity, (d) inhibits C3 cleavage, (e) inhibits C3 cleavage and C3a release, (f) inhibits complement activation, (g) inhibits activation of the alternative complement pathway, (h) inhibits activation of the classical complement pathway, (i) inhibits activation of the alternative complement pathway and inhibits activation of the classical complement pathway, (j) does not detectably bind to Factor Bb, (k) does not detectably bind to C3d, and (1) does not detectably bind to C3a. In certain embodiments, the C3 -binding agents inhibit activation of the classical complement pathway. In some embodiments, the C3-binding agents inhibit activation of the alternative complement pathway. In some embodiments, the C3 -binding agents inhibit activation of the alternative complement pathway and classical complement pathway.
[0027] In some embodiments, a C3-binding agent (e.g., antibody) comprises: (1) a heavy chain variable region (VH) comprising VH complementarity determining region (CDR)1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:31, and a light chain variable region (VL) comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:32; (2) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 33, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (3) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:34, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (4) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:222, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (5) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:223, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (6) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:224, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35; (7) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 77, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:78; (8) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:95, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:96; (9) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:l 13, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 114; (10) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 131, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 132; (11) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 149, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:150; (12) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 167, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 168; (13) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 185, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 186; (14) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:203, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:204; or (15) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:220, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:221.
[0028] In some embodiments, a C3 -binding agent (e.g., antibody) comprises (a) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 16, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:20, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (b) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:25, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 16, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:25, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:26, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (c) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:27, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:28, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:29, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:27, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:30, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (d) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:48, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:49, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:50, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:48, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:51, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (e) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 52, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:53, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 54, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:52, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:55, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (f) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:56, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:57, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 58, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:56, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:59, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (g) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:60, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:64, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:75; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:61, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:65, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:75; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:60, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:67, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:75; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:62, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:64, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:75; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:63, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:68, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:70, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:72, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:74, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:76; (h) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:79, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:83, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:93; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:80, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:84, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:93; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:79, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:85, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:93; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:81, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:83, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:93; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:82, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:86, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:88, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:90, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:92, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:94; (i) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:97, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 111; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:98, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 102, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 111; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:97, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 103, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:l 11; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:99, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:l 11; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 100, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 104, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 106, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 108, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:l 10, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 112; (j) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 115, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 115, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 121, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 117, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 118, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 122, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 126, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 130; (k) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 133, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 138, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 133, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 139, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 135, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 136, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 140, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 142, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 144, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 146, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 148; (1) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:151, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 156, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:151, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 157, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 153, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 154, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 158, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 160, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 162, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 164, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 166; (m) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 169, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 173, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 170, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 174, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 169, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 175, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 171, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 173, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 171, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 172, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 176, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 178, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 180, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 182, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 184; (n) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 187, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 191, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 188, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 192, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 187, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 193, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 189, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 191, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 190, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 194, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:200, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:202; or (o) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:205, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:209, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218; (ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:206, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:210, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218; (iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:205, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:211, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218; (iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:207, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:209, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218; or (v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:208, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:212, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 214, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:215, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:217, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:219.
[0029] In some embodiments of the C3-binding agent (e.g., antibody), (a) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31; (b) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:33; (c) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:34; (d) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222; (e) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223; (f) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224; (g) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77; (h) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:95; (i) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 113; (j) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131; (k) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149; (1) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167; (m) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185; (n) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203; (o) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220; (p) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32; (q) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (r) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78; (s) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:96; (t) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 114; (u) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132; (v) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150; (w) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168; (x) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186; (y) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204; (z) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221; (aa) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 31 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32; (bb) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (cc) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (dd) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (ee) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (ff) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35; (gg) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78; (hh) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:95 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 96; (ii) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 113 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 114; (jj) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132; (kk) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150; (11) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168; (mm) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186; (nn) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204; or (oo) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221.
[0030] In some embodiments of the C3-binding agent (e.g., antibody), (a) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31; (b) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33; (c) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:34; (d) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222; (e) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223; (f) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224; (g) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77; (h) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:95; (i) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 113; (j) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131; (k) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149; (1) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167; (m) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185; (n) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203; (o) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220; (p) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32; (q) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (r) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78; (s) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:96; (t) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114; (u) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132; (v) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150; (w) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168; (x) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186; (y) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204; (z) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221; (aa) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 31 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32; (bb) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (cc) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (dd) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (ee) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (ff) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35; (gg) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78; (hh) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:95 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 96; (ii) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 113 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114; (jj) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132; (kk) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150; (11) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168; (mm) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186; (nn) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204; or (oo) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221.
[0031] In some embodiments of the C3-binding agent (e.g., antibody), (a) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:31; (b) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33; (c) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34; (d) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:222; (e) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:223; (f) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:224; (g) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:77; (h) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:95; (i) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 113; (j) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 131; (k) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 149; (1) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 167; (m) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 185; (n) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:203; (o) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:220; (p) the light chain variable region comprises amino acid sequence of SEQ ID NO:32; (q) the light chain variable region comprises amino acid sequence of SEQ ID NO:35; (r) the light chain variable region comprises amino acid sequence of SEQ ID NO:78; (s) the light chain variable region comprises amino acid sequence of SEQ ID NO:96; (t) the light chain variable region comprises amino acid sequence of SEQ ID NO: 114; (u) the light chain variable region comprises amino acid sequence of SEQ ID NO: 132; (v) the light chain variable region comprises amino acid sequence of SEQ ID NO: 150; (w) the light chain variable region comprises amino acid sequence of SEQ ID NO: 168; (x) the light chain variable region comprises amino acid sequence of SEQ ID NO: 186; (y) the light chain variable region comprises amino acid sequence of SEQ ID NO:204; (z) the light chain variable region comprises amino acid sequence of SEQ ID NO:221; (aa) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:31 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:32; (bb) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (cc) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (dd) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (ee) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (ff) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (gg) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:78; (hh) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:95 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:96; (ii) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 113 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 114; (jj) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 132; (kk) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 150; (11) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 168; (mm) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 186; (nn) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:204; or (oo) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:221.
[0032] In some embodiments, a C3-binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3- binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprises a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO: 52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO:52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3- binding agent comprises a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNTGTTYNQQFTG (SEQ ID NO:48), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHEGGTTYNQQFTG (SEQ ID NO:52), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, the agent is an antibody. In some embodiments, the C3-binding agent is an anti-C3 antibody. The present disclosure also provides an C3-binding agent that comprises a heavy chain variable region comprising the three VH CDRs according to any CDR definition provided in Table 1 A (for 38G10), Table IB (for Hz38G10 and Hz38G10(G56A)), and Table 1C (for Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q)) and a light chain variable region comprising the three VL CDRs according to any CDR definition provided in Table 1A (for 38G10), Table IB (for Hz38G10 and Hz38G10(G56A)), and Table 1C (for Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q)).
[0033] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to any one of SEQ ID NO: 31, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, and SEQ ID NO:224; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:32 or SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:31 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:32. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:33 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:34 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:222 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:223 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:224 and a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region comprising SEQ ID NO:31 and a light chain variable region comprising SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:33 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:34 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:222 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:223 and a light chain variable region comprising SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising SEQ ID NO:224 and a light chain variable region comprising SEQ ID NO:35.
[0034] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from: (a) the antibody designated 38G10 that comprises a heavy chain variable region comprising SEQ ID NO:31 and a light chain variable region comprising SEQ ID NO:32; (b) the antibody designated Hz38G10 that comprises a heavy chain variable region comprising SEQ ID NO:33 and a light chain variable region comprising SEQ ID NO:35; (c) the antibody designated Hz38G10(G56A) that comprises a heavy chain variable region comprising SEQ ID NO:34 and a light chain variable region comprising SEQ ID NO:35; (d) the antibody designated Hz38G10 (G56T) that comprises a heavy chain variable region comprising SEQ ID NO:222 and a light chain variable region comprising SEQ ID NO:35; (e) the antibody designated Hz38G10 (N55E) that comprises a heavy chain variable region comprising SEQ ID NO:223 and a light chain variable region comprising SEQ ID NO:35; or (f) the antibody designated Hz38G10 (N55Q) that comprises a heavy chain variable region comprising SEQ ID NO:224 and a light chain variable region comprising SEQ ID NO:35.
[0035] In some embodiments, the C3-binding agent is an anti-C3 antibody.
[0036] In some embodiments, a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:37 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and/or a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:39 and a light chain having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:41.
[0037] In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41.
In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an anti-C3 antibody that comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41.
[0038] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 3D8 that comprises a heavy chain variable region comprising SEQ ID NO:77 and a light chain variable region comprising SEQ ID NO:78. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:77 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:78. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 2.
[0039] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:77; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:78. In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:77; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:78. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:77; and a light chain variable region having the sequence of SEQ ID NO:78.
[0040] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 3G8 that comprises a heavy chain variable region comprising SEQ ID NO:95 and a light chain variable region comprising SEQ ID NO:96. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 95 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:96. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 3.
[0041] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:95; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:96. In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:95; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:96. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:95; and a light chain variable region having the sequence of SEQ ID NO:96.
[0042] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 1502 that comprises a heavy chain variable region comprising SEQ ID NO: 113 and a light chain variable region comprising SEQ ID NO: 114. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 113 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:l 14. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 4.
[0043] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 113; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:l 14. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 113; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 113; and a light chain variable region having the sequence of SEQ ID NO: 114. [0044] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 27A8 that comprises a heavy chain variable region comprising SEQ ID NO: 131 and a light chain variable region comprising SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 131 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO: 132. In some embodiments, the C3-binding agent is an anti-C3 antibody. For example, a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 5.
[0045] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 131; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 132. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 131; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 131; and a light chain variable region having the sequence of SEQ ID NO: 132. [0046] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 28C3 that comprises a heavy chain variable region comprising SEQ ID NO: 149 and a light chain variable region comprising SEQ ID NO: 150. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 149 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:150. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 6.
[0047] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 149; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 150. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 149; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 150. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 149; and a light chain variable region having the sequence of SEQ ID NO: 150. [0048] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 38F5 that comprises a heavy chain variable region comprising SEQ ID NO: 167 and a light chain variable region comprising SEQ ID NO: 168. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO: 167 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO: 168. In some embodiments, the C3-binding agent is an anti-C3 antibody. For example, a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 7.
[0049] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 167; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 168. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 167; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 168. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 167; and a light chain variable region having the sequence of SEQ ID NO: 168. [0050] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 62B11 that comprises a heavy chain variable region comprising SEQ ID NO: 185 and a light chain variable region comprising SEQ ID NO: 186. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VEl CDR2, and VH CDR3 of SEQ ID NO: 185 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:186. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 8.
[0051] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 185; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 186. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 185; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO: 185; and a light chain variable region having the sequence of SEQ ID NO: 186. [0052] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 62F2 that comprises a heavy chain variable region comprising SEQ ID NO:203 and a light chain variable region comprising SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:203 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:204. In some embodiments, the C3-binding agent is an anti-C3 antibody. For example, a C3- binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 9.
[0053] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:203; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:204. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:203; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:203; and a light chain variable region having the sequence of SEQ ID NO:204. [0054] In some embodiments, a C3-binding agent comprises the heavy chain CDR1, CDR2, and CDR3, and/or the light chain CDR1, CDR2, and CDR3 from the antibody designated 63 A3 that comprises a heavy chain variable region comprising SEQ ID NO:220 and a light chain variable region comprising SEQ ID NO:221. In some embodiments, the C3-binding agent is an anti-C3 antibody. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the VH CDR1, VH CDR2, and VH CDR3 of SEQ ID NO:220 and a light chain variable region comprising the VL CDR1, VL CDR2, and VL CDR3 of SEQ ID NO:221. For example, a C3-binding agent can comprise the six CDRs of any of the CDR definitions provided in Table 10.
[0055] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:220; and/or (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:221. In some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:220; and (b) a light chain variable region having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100%) sequence identity to SEQ ID NO:221. In some embodiments, a C3-binding agent comprises a heavy chain variable region having the sequence of SEQ ID NO:220; and a light chain variable region having the sequence of SEQ ID NO:221. [0056] In some embodiments of each of the aforementioned aspects and embodiments, as well as other aspects and embodiments described herein, a C3 -binding agent is an antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a whole or intact antibody. In some embodiments, the antibody is an IgG antibody. In some embodiments, the antibody is an IgGl antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the antibody is a human IgGl antibody, a human IgG2 antibody, a human IgG3 antibody, or a human IgG4 antibody. In some embodiments, the antibody comprises a human kappa chain or a human lambda light chain constant region. In some embodiments, the antibody is a bispecific antibody or a multispecific antibody. In some embodiments, the antibody is an antibody fragment. In some embodiments, the antibody or antibody fragment is a Fab, Fab’, F(ab’)2, Fv, scFv, (scFv)2, single chain antibody, dual variable region antibody, single variable region antibody, linear antibody, nanobody, or a V region antibody.
[0057] In some embodiments, a C3-binding agent (e.g, an antibody) is an scFv. In some embodiments, the scFv comprises a heavy chain variable region comprising SEQ ID NO:225 or SEQ ID NO:226. In some embodiments, the scFv comprises a light chain variable region comprising SEQ ID NO:227. In some embodiments, the scFv comprises a heavy chain variable region comprising SEQ ID NO:225 and a light chain variable region comprising SEQ ID NO:227. In some embodiments, the scFv comprises a heavy chain variable region comprising SEQ ID NO:226 and a light chain variable region comprising SEQ ID NO:227. In some embodiments, the scFv comprises SEQ ID NO:228. In some embodiments, the scFv comprises SEQ ID NO:229. In some embodiments, the scFv comprises SEQ ID NO:230. In some embodiments, the scFv comprises SEQ ID NO:231.
[0058] In some embodiments of each of the aforementioned aspects and embodiments, as well as other aspects and embodiments described herein, the anti-C3 antibody is an antagonistic antibody. In some embodiments, the anti-C3 antibody inhibits C3 activity. In some embodiments, the anti-C3 antibody inhibits activation of the complement system. In some embodiments, the anti-C3 antibody inhibits activation of the classical complement pathway. In some embodiments, the anti-C3 antibody inhibits activation of the alternative complement pathway. In some embodiments, the anti-C3 antibody inhibits activation of the classical complement pathway and the alternative complement pathway. [0059] In another aspect, the disclosure provides compositions comprising a C3- binding agent described herein. In some embodiments, the disclosure provides compositions comprising an anti-C3 antibody described herein.
[0060] In another aspect, the disclosure provides pharmaceutical compositions comprising a C3-binding agent described herein and a pharmaceutically acceptable carrier. In some embodiments, the disclosure provides pharmaceutical compositions comprising an anti-C3 antibody described herein and a pharmaceutically acceptable carrier.
[0061] In some embodiments of each of the aforementioned aspects, as well as other aspects and/or embodiments described elsewhere herein, the C3-binding agent is isolated. In some embodiments, the C3-binding agent is substantially pure.
[0062] In another aspect, the disclosure provides polynucleotides comprising a polynucleotide or polynucleotides that encode a C3 -binding agent described herein. In some embodiments, the polynucleotide or polynucleotides are isolated. In some embodiments, a vector or vectors comprise the polynucleotide or polynucleotides that encode a C3-binding agent described herein. In some embodiments, an isolated cell comprises the polynucleotide or polynucleotides that encode a C3-binding agent described herein. In some embodiments, an isolated cell comprises the vector or vectors comprising a polynucleotide or polynucleotides that encode a C3-binding agent described herein. In some embodiments, the disclosure provides a cell comprising a C3-binding agent described herein. In some embodiments, the disclosure provides a cell producing a C3 -binding agent described herein. In some embodiments, a cell produces an anti-C3 antibody described herein. In some embodiments, a cell is a monoclonal cell line. In some embodiments, a cell is a hybridoma.
[0063] Where aspects or embodiments of the disclosure are described in terms of a Markush group or other grouping of alternatives, the present disclosure encompasses not only the entire group listed as a whole, but also each member of the group individually and all possible subgroups of the main group, and also the main group absent one or more of the group members. The present disclosure also envisages the explicit exclusion of one or more of any of the group members in the claimed disclosure.
BRIEF DESCRIPTION OF THE FIGURES [0064] Figure 1. C3a release assay.
[0065] Figure 2. Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody 38G10, Ab2, and Ab3 were tested for their ability to inhibit complement activation.
[0066] Figure 3. Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody Hz38G10 and compstatin derivative (COMP) were tested for their ability to inhibit complement activation.
[0067] Figure 4. Alternative Pathway and Classical Pathway Hemolysis Assays. Antibody Hz38G10 and a pegylated version of the compstatin derivative (COMP -PEG) were tested for their ability to inhibit complement activation.
[0068] Figure 5. Binding Site Analysis. Hz38G10 and a compstatin derivative were tested in a competitive binding assay to assess whether the molecules bind to the same or different epitopes.
DETAILED DESCRIPTION
[0069] The present disclosure provides methods of treating respiratory illness, methods of preventing or inhibiting the development of respiratory illness, methods of inhibiting complement pathway activation, methods of treating thrombosis, methods of preventing or inhibiting the development of thrombosis, and methods of treating a coronavirus infection or the resulting complications and/or sequelae of a coronavirus infection. The methods provided herein comprise administering to a subject agents that specifically bind human complement component C3 (i.e., a C3-binding agent). In some embodiments, the C3 -binding agents include, but are not limited to, polypeptides, antibodies and antigen-binding fragments thereof, scaffold proteins, and heterodimeric molecules. C3-binding agents include, but are not limited to, antagonists of C3 activity, inhibitors of C3 activity, and/or agents that modulate C3 activity. Related polypeptides, polynucleotides, vectors, compositions comprising the agents, cells comprising the related polynucleotides or vectors, and methods of making the agents are also provided and/or used in the methods described herein.
I. Definitions
[0070] Unless otherwise defined herein, technical and scientific terms used in the present description have the meanings that are commonly understood by those of ordinary skill in the art. For purposes of interpreting this specification, the following description of terms will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. In the event that any description of a term set forth conflicts with any document incorporated herein by reference, the description of the term set forth below shall control.
[0071] The term “binding agent” as used herein refers to a molecule which binds a specific antigen or target ( e.g ., complement component C3). A binding agent may comprise a protein, peptide, nucleic acid, carbohydrate, lipid, or small molecular weight compound. In some embodiments, a binding agent comprises an antibody. In some embodiments, a binding agent is an antibody. In some embodiments, a binding agent comprises an alternative protein scaffold or artificial scaffold (e.g., a nonimmunoglobulin backbone). In some embodiments, a binding agent is a fusion protein comprising an antigen-binding site. In some embodiments, a binding agent is a bispecific or multispecific molecule comprising at least one antigen-binding site.
[0072] The term “antibody” as used herein refers to an immunoglobulin molecule, or antigen-binding fragment thereof, that recognizes and binds a target through at least one antigen-binding site. “Antibody” is used herein in the broadest sense and encompasses various antibody structures, including but not limited to, polyclonal antibodies, recombinant antibodies, monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, bispecific antibodies, multispecific antibodies, diabodies, tribodies, tetrabodies, single chain Fv (scFv) antibodies, and antibody fragments as long as they exhibit the desired antigen-binding activity. [0073] The term “intact antibody” or “full-length antibody” refers to an antibody having a structure substantially similar to a native antibody structure. This includes, for example, an antibody comprising two light chains each comprising a variable region and a light chain constant region (CL) and two heavy chains each comprising a variable region and at least a hinge region and heavy chain constant regions CHI, CH2, and CH3.
[0074] The term “antibody fragment” as used herein refers to a molecule other than an intact antibody that comprises a portion of an antibody and generally at least one antigen-binding site. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, single chain antibody molecules (e.g, scFv), disulfide-linked scFv (dsscFv), nanobodies, diabodies, tribodies, tetrabodies, minibodies, dual variable domain antibodies (DVD), single variable domain antibodies (e.g, camelid antibodies), and bispecific or multispecific molecules formed from antibody fragments.
[0075] The term “monoclonal antibody” as used herein refers to a substantially homogenous antibody population involved in the highly specific recognition and binding of a single antigenic determinant or epitope. The term “monoclonal antibody” encompasses intact and full-length monoclonal antibodies as well as antibody fragments (e.g, Fab, Fab', F(ab')2, Fv), single chain antibodies (e.g, scFv), fusion proteins comprising an antibody fragment, and any other modified immunoglobulin molecule comprising at least one antigen-binding site. Furthermore, “monoclonal antibody” refers to such antibodies made by any number of techniques, including but not limited to, hybridoma production, phage library display, recombinant expression, and transgenic animals.
[0076] The term “chimeric antibody” refers to an antibody in which a portion of the heavy and/or light chain is derived from a first source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
[0077] The term “humanized antibody” as used herein refers to an antibody that comprises a human heavy chain variable region and a light chain variable region wherein the native CDR residues are replaced by residues from corresponding CDRs from a nonhuman antibody (e.g, mouse, rat, rabbit, or nonhuman primate), wherein the nonhuman antibody has the desired specificity, affinity, and/or activity. In some embodiments, one or more framework region residues of the human heavy chain or light chain variable regions are replaced by corresponding residues from nonhuman antibody. Furthermore, humanized antibodies can comprise residues that are not found in the human antibody or in the nonhuman antibody. In some embodiments, these modifications are made to further refine and/or optimize antibody characteristics. In some embodiments, the humanized antibody comprises at least a portion of an immunoglobulin constant region ( e.g ., hinge region, CHI, CH2, CH3, and/or Fc), typically that of a human immunoglobulin.
[0078] The term “human antibody” as used herein refers to an antibody that possesses an amino acid sequence that corresponds to an antibody produced by a human and/or an antibody that has been made using any of the techniques that are known to those of skill in the art for making human antibodies. These techniques include, but not limited to, phage display libraries, yeast display libraries, transgenic animals, recombinant protein production, and B-cell hybridoma technology.
[0079] The terms “epitope” and “antigenic determinant” are used interchangeably herein and refer to that portion of an antigen or target capable of being recognized and bound by a particular antibody. When the antigen or target is a polypeptide, epitopes can be formed both from contiguous amino acids and noncontiguous amino acids juxtaposed by tertiary folding of the protein. Epitopes formed from contiguous amino acids (also referred to as linear epitopes) are typically retained upon protein denaturing, whereas epitopes formed by tertiary folding (also referred to as conformational epitopes) are typically lost upon protein denaturing. An epitope typically includes at least 3, and more usually, at least 5, 6, 7, or 8-10 amino acids in a unique spatial conformation. Epitopes can be predicted using any one of a large number of software bioinformatic tools available on the internet. X-ray crystallography may be used to characterize an epitope on a target protein by analyzing the amino acid residue interactions of an antigen/antibody complex.
[0080] The term “specifically binds” as used herein refers to an agent (e.g., an antibody) that interacts more frequently, more rapidly, with greater duration, with greater affinity, or with some combination of the above to a particular antigen, epitope, protein, or target molecule than with alternative substances. A binding agent that specifically binds an antigen can be identified, for example, by immunoassays, ELISAs, SPR ( e.g ., Biacore), or other techniques known to those of skill in the art. In some embodiments, an agent that specifically binds an antigen (e.g., human C3) can bind related antigens (e.g, cyno C3). A binding agent that specifically binds an antigen can bind the target antigen at a higher affinity than its affinity for a different antigen. The different antigen can be a related antigen. In some embodiments, a binding agent that specifically binds an antigen can bind the target antigen with an affinity that is at least 20 times greater, at least 30 times greater, at least 40 times greater, at least 50 times greater, at least 60 times greater, at least 70 times greater, at least 80 times greater, at least 90 times greater, or at least 100 times greater, than its affinity for a different antigen. In some embodiments, a binding agent that specifically binds a particular antigen binds a different antigen at such a low affinity that binding cannot be detected using an assay described herein or otherwise known in the art. In some embodiments, affinity is measured using SPR technology in a Biacore system as described herein or as known to those of skill in the art.
[0081] The terms “polypeptide” and “peptide” and “protein” are used interchangeably herein and refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by nonamino acids. The terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification.
Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid, including but not limited to, unnatural amino acids, as well as other modifications known in the art. It is understood that, because the polypeptides of this disclosure may be based upon antibodies, the term “polypeptide” encompasses polypeptides as a single chain and polypeptides of two or more associated chains. [0082] The terms “polynucleotide” and “nucleic acid” and “nucleic acid molecule” are used interchangeably herein and refer to polymers of nucleotides of any length, and include DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase.
[0083] The terms “identical” or percent “identity” in the context of two or more nucleic acids or polypeptides, refer to two or more sequences or subsequences that are the same or have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned (introducing gaps, if necessary) for maximum correspondence, not considering any conservative amino acid substitutions as part of the sequence identity. The percent identity may be measured using sequence comparison software or algorithms or by visual inspection. Various algorithms and software that may be used to obtain alignments of amino acid or nucleotide sequences are well- known in the art. These include, but are not limited to, BLAST, ALIGN, Megalign, BestFit, GCG Wisconsin Package, and variants thereof. In some embodiments, two nucleic acids or polypeptides of the disclosure are substantially identical, meaning they have at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, and in some embodiments at least 95%, 96%, 97%, 98%, 99% nucleotide or amino acid residue identity, when compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection. In some embodiments, identity exists over a region of the sequences that is at least 10, at least 20, at least 20- 40, at least 40-60 nucleotides or amino acid residues, at least 60-80 nucleotides or amino acid residues in length or any integral value there between. In some embodiments, identity exists over a longer region than 60-80 nucleotides or amino acid residues, such as at least 80-100 nucleotides or amino acid residues, and in some embodiments the sequences are substantially identical over the full length of the sequences being compared, for example, (i) the coding region of a nucleotide sequence or (ii) an amino acid sequence.
[0084] The phrase “conservative amino acid substitution” as used herein refers to a substitution in which one amino acid residue is replaced with another amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been generally defined in the art, including basic side chains ( e.g ., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g, glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g, threonine, valine, isoleucine) and aromatic side chains (e.g, tyrosine, phenylalanine, tryptophan, histidine). For example, substitution of a valine for a leucine is considered to be a conservative substitution. Generally, conservative substitutions in the sequences of polypeptides and/or antibodies do not abrogate the binding of the polypeptide or antibody to the target binding site. Methods of identifying nucleotide and amino acid conservative substitutions that do not eliminate binding are well-known in the art.
[0085] The term “vector” as used herein means a construct, which is capable of delivering, and usually expressing, one or more gene(s) or sequence(s) of interest in a host cell. Examples of vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid, or phage vectors, DNA or RNA expression vectors associated with cationic condensing agents, and DNA or RNA expression vectors encapsulated in liposomes.
[0086] The term “isolated” as used herein refers to a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition which is in a form not found in nature. An “isolated” antibody is substantially free of material from the cellular source from which it is derived. In some embodiments, isolated polypeptides, soluble proteins, antibodies, polynucleotides, vectors, cells, or compositions are those which have been purified to a degree that they are no longer in a form in which they are found in nature. In some embodiments, a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition which is isolated is substantially pure. A polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition may be isolated from a natural source (e.g, tissue) or from a source such as an engineered cell line. [0087] The term “substantially pure” as used herein refers to material which is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 98% pure, or at least 99% pure.
[0088] The term “subject” refers to any animal ( e.g ., a mammal), including, but not limited to, humans, non-human primates, canines, felines, rabbits, rodents, and the like. [0089] The term “pharmaceutically acceptable” as used herein refers to a substance approved or approvable by a regulatory agency or listed in the U.S. Pharmacopeia, European Pharmacopeia, or other generally recognized pharmacopeia for use in animals, including humans.
[0090] The terms “pharmaceutically acceptable excipient, carrier, or adjuvant” or “acceptable pharmaceutical carrier” as used herein refer to an excipient, carrier, or adjuvant that can be administered to a subject (e.g., a human), together with at least one therapeutic agent and which is generally safe, non-toxic, and has no effect on the pharmacological activity of the therapeutic agent. In general, those of skill in the art and governmental agencies consider a pharmaceutically acceptable excipient, carrier, or adjuvant to be an inactive ingredient of any formulation.
[0091] The term “pharmaceutical formulation” or “pharmaceutical composition” as used herein refers to a preparation which is in such form as to permit the biological activity of the agent to be effective. A pharmaceutical formulation or composition generally comprises additional components, such as a pharmaceutically acceptable excipient, carrier, adjuvant, buffers, etc.
[0092] The term “effective amount” or “therapeutically effective amount” as used herein refers to the amount of an agent which is sufficient to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder or condition in a subject, and/or (ii) a symptom in a subject. The term also encompasses an amount of an agent necessary for the (i) reduction or amelioration of the advancement or progression of a given disease, disorder, or condition, (ii) reduction or amelioration of the recurrence, development, or onset of a given disease, disorder, or condition, and/or (iii) the improvement or enhancement of the prophylactic or therapeutic effect(s) of another agent or therapy (e.g, an agent other than the binding agents provided herein). [0093] The term “therapeutic effect” as used herein refers to the effect and/or ability of an agent to reduce and/or ameliorate the severity and/or duration of (i) a disease, disorder, or condition in a subject, and/or (ii) a symptom in a subject. The term also encompasses the ability of an agent to (i) reduce or ameliorate the advancement or progression of a given disease, disorder, or condition, (ii) reduce or ameliorate the recurrence, development, or onset of a given disease, disorder, or condition, and/or (iii) to improve or enhance the prophylactic or therapeutic effect(s) of another agent or therapy ( e.g an agent other than the binding agents provided herein).
[0094] The term “treat” or “treatment” or “treating” or “to treat” or “alleviate” or alleviation” or “alleviating” or “to alleviate” as used herein refers to therapeutic measures that aim to cure, slow down, lessen symptoms of, and/or halt progression of a pathologic condition or disorder.
[0095] The term “prevent” or “prevention” or “preventing” as used herein refers to the partial or total inhibition of the development, recurrence, onset, or spread of a disease, disorder, or condition, or a symptom thereof in a subject.
[0096] As used herein, reference to “about” or “approximately” a value or parameter includes (and describes) embodiments that are directed to that value or parameter. For example, a description referring to “about X” includes description of “X”.
[0097] As used in the present disclosure and claims, the singular forms “a”, “an” and “the” include plural forms unless the context clearly dictates otherwise.
[0098] It is understood that wherever embodiments are described herein with the term “comprising” otherwise analogous embodiments described in terms of “consisting of’ and/or “consisting essentially of’ are also provided. It is also understood that wherever embodiments are described herein with the phrase “consisting essentially of’ otherwise analogous embodiments described in terms of “consisting of’ are also provided.
[0099] The term “and/or” as used in a phrase such as “A and/or B” herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term “and/or” as used in a phrase such as “A, B, and/or C” is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). II. C3-binding agents
[00100] Amino acid (aa) sequences for human C3 (UniProtKB No. P01024) and cynomolgus monkey (“cyno”) (NCBI Ref No. XP_005587776.1) are provided herein as SEQ ID NO: 1 and SEQ ID NO:7, respectively. As used herein, reference to amino acid positions of C3 refer to the numbering of amino acid sequences including the signal sequence.
[00101] The present disclosure provides C3-binding agents for use in the methods described herein. In some embodiments, the C3 -binding agent binds human C3 and has at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12) of the following properties: (a) binds to cyno C3, (b) is an antagonist of C3, (c) inhibits C3 activity, (d) inhibits C3 cleavage, (e) inhibits C3 cleavage and C3a release, (f) inhibits complement activation, (g) inhibits activation of the alternative complement pathway, (h) inhibits activation of the classical complement pathway, (i) inhibits activation of the alternative complement pathway and inhibits activation of the classical complement pathway, (j) does not detectably bind to Factor Bb, (k) does not detectably bind to C3d, and (1) does not detectably bind to C3a. In certain embodiments, the C3 -binding agent inhibits activation of the classical complement pathway. In some embodiments, the C3-binding agent inhibits activation of the alternative complement pathway and classical complement pathway. In some embodiments, a C3-binding agent binds C3. In some embodiments, a C3-binding agent binds a fragment of C3. In some embodiments, a C3- binding agent binds within a specific region of C3. In some embodiments, a C3-binding agent binds an epitope on C3. In some embodiments, a C3-binding agent binds a linear epitope on C3. In some embodiments, a C3-binding agent binds a conformational epitope on C3. In some embodiments, a C3-binding agent binds human C3. In some embodiments, a C3 -binding agent binds cyno C3. In some embodiments, a C3 -binding agent binds human C3 and cyno C3. In some embodiments, a C3-binding agent binds C3 and does not bind C3b. In some embodiments, a C3 -binding agent binds C3 and does not bind C3b at a detectable level. [00102] Assays for determining inhibition of C3 cleavage and C3a release are known to those skilled in the art. In certain embodiments, the C3 -binding agent inhibits C3 cleavage and C3a release at a concentration of 0.1 to 1000 nM. In certain embodiments, a C3-binding agent inhibits C3 cleavage and C3a release if there is a reduction by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% in the amount of C3 cleavage and C3a release compared to C3 cleavage and C3a release in the absence of the C3-binding agent.
[00103] Assays for determining whether a C3 -binding agent inhibits activation of the alternative complement pathway or the classical complement pathway are known to those skilled in the art.
[00104] Assays for determining the binding of a C3 -binding agent to factor Bb, C3d, or C3a are known in the art. In certain embodiments, a C3-binding agent is determined to not detectably bind to factor Bb, C3d, or C3a if the C3 -binding agent does not bind to factor Bb, C3d, or C3a, respectively, at a concentration of up to 1 mM according to an assay described herein ( e.g ., BiaCore).
[00105] In some embodiments, a C3-binding agent is an antibody. In some embodiments, the antibody is a recombinant antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In some embodiments, the antibody is an IgGl antibody. In some embodiments, the antibody is an IgG2 antibody. In some embodiments, the antibody is an IgG3 antibody. In some embodiments, the antibody is an IgG4 antibody. In some embodiments, the antibody is a human IgGl antibody. In some embodiments, the antibody is a human IgG2 antibody. In some embodiments, the antibody is a human IgG3 antibody. In some embodiments, the antibody is a human IgG4 antibody. In some embodiments, the antibody comprises a human kappa light chain constant region. In some embodiments, the antibody comprises a human lambda light chain constant region. In some embodiments, the antibody is an antibody fragment comprising an antigen-binding site. In some embodiments, the antibody is a scFv. In some embodiments, the antibody is a disulfide-linked scFv. In some embodiments, the antibody is a bispecific antibody or a multispecific antibody. In some embodiments, the antibody is a monovalent antibody. In some embodiments, the antibody is a monospecific antibody. In some embodiments, the antibody is a bivalent antibody.
[00106] In some embodiments, the antibody is isolated. In some embodiments, the antibody is substantially pure.
[00107] In some embodiments, a C3-binding agent is a polyclonal antibody.
Polyclonal antibodies can be prepared by any method known to those of skill in the art. In some embodiments, polyclonal antibodies are produced by immunizing an animal ( e.g ., a rabbit, rat, mouse, goat, donkey) with an antigen of interest (e.g, a purified peptide fragment, a recombinant protein, or a fusion protein) using multiple subcutaneous or intraperitoneal injections. In some embodiments, the antigen is conjugated to a carrier such as keyhole limpet hemocyanin (KLH), serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor. The antigen (with or without a carrier protein) is diluted in sterile saline and usually combined with an adjuvant (e.g, Complete or Incomplete Freund's Adjuvant) to form a stable emulsion. After a period of time, polyclonal antibodies are recovered from the immunized animal (e.g, from blood or ascites). In some embodiments, the polyclonal antibodies are purified from serum or ascites according to standard methods in the art including, but not limited to, affinity chromatography, ion-exchange chromatography, gel electrophoresis, and/or dialysis.
[00108] In some embodiments, a C3-binding agent is a monoclonal antibody. Monoclonal antibodies can be prepared by any method known to those of skill in the art. In some embodiments, monoclonal antibodies are prepared using hybridoma methods known to one of skill in the art. For example, using a hybridoma method, a mouse, rat, rabbit, hamster, or other appropriate host animal, is immunized as described above. In some embodiments, lymphocytes are immunized in vitro. In some embodiments, the immunizing antigen is a human protein or a fragment thereof. In some embodiments, the immunizing antigen is a mouse protein or a fragment thereof. [00109] Following immunization, lymphocytes are isolated and fused with a suitable myeloma cell line using, for example, polyethylene glycol. The hybridoma cells are selected using specialized media as known in the art and unfused lymphocytes and myeloma cells do not survive the selection process. Hybridomas that produce monoclonal antibodies directed specifically against a chosen antigen can be identified by a variety of methods including, but not limited to, immunoprecipitation, immunoblotting, and in vitro binding assays (e.g, flow cytometry, FACS, ELISA, SPR ( e.g. , Biacore), and radioimmunoassay). Once hybridoma cells that produce antibodies of the desired specificity, affinity, and/or activity are identified, the clones may be subcloned by limiting dilution techniques. The hybridomas can be propagated either in in vitro culture using standard methods or in vivo as ascites tumors in an animal. The monoclonal antibodies can be purified from the culture medium or ascites fluid according to standard methods in the art including, but not limited to, affinity chromatography, ion-exchange chromatography, gel electrophoresis, and dialysis. [00110] In some embodiments, C3-binding agents (e.g., monoclonal antibodies) are made using recombinant DNA techniques as known to one skilled in the art. For example, a polynucleotide or polynucleotides encoding an antibody are isolated from mature B-cells or hybridoma cells, such as by RT-PCR using oligonucleotide primers that specifically amplify the genes encoding the heavy and light chains of the antibody, and their sequence is determined using standard techniques. The isolated polynucleotides encoding the heavy and light chains are then cloned into suitable expression vectors which produce the monoclonal antibodies when transfected into host cells such as E. coli , simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins.
[00111] In some embodiments, a C3-binding agent is an antibody that is made using recombinant DNA techniques as known to one skilled in the art. For example, a polynucleotide or polynucleotides encoding the heavy and light chains thenare cloned into suitable expression vectors which produce the antibodies when transfected into host cells such as E. coli, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins.
[00112] In some embodiments, recombinant antibodies (e.g., monoclonal antibodies) are isolated from phage display libraries expressing variable domains or CDRs of a desired species. Screening of phage libraries can be accomplished by various techniques known in the art.
[00113] In some embodiments, a C3-binding agent (e.g., a monoclonal antibody) is modified by using recombinant DNA technology to generate alternative antibodies. In some embodiments, the constant domains of the light chain and heavy chain of a mouse monoclonal antibody are substituted for constant regions of a human antibody to generate a chimeric antibody. In some embodiments, the constant regions are truncated or removed to generate a desired antibody fragment of a monoclonal antibody. In some embodiments, site-directed or high-density mutagenesis of the variable region(s) is used to optimize specificity and affinity of a monoclonal antibody.
[00114] In some embodiments, a C3-binding agent is a humanized antibody. Various methods for generating humanized antibodies are known in the art. In some embodiments, a humanized antibody comprises one or more amino acid residues that have been introduced into it from a source that is non-human. In some embodiments, humanization is performed by substituting one or more non-human CDR sequences for the corresponding CDR sequences of a human antibody. In some embodiments, the humanized antibodies are constructed by substituting all six CDRs of a non-human antibody (e.g., a mouse antibody) for the corresponding CDRs of a human antibody. [00115] The choice of which human heavy chain variable region and/or light chain variable region is used for generating humanized antibodies can be made based on a variety of factors and by a variety of methods known in the art. In some embodiments, the “best-fit” method is used where the sequence of the variable region of a non-human (e.g, rodent) antibody is screened against the entire library of known human variable region sequences. The human sequence that is most similar to that of the non-human (e.g, rodent) sequence is selected as the human variable region framework for the humanized antibody. In some embodiments, a particular variable region framework derived from a consensus sequence of all human antibodies of a particular subgroup of light or heavy chains is selected as the variable region framework. In some embodiments, the variable region framework sequence is derived from the consensus sequences of the most abundant human subclasses. In some embodiments, human germline genes are used as the source of the variable region framework sequences. [00116] Other methods for humanization include, but are not limited to, a method called “superhumanization” which is described as the direct transfer of CDRs to a human germline framework, a method termed Human String Content (HSC) which is based on a metric of “antibody humanness”, methods based on generation of large libraries of humanized variants (including phage, ribosomal, and yeast display libraries), and methods based on framework region shuffling.
[00117] In some embodiments, a C3-binding agent is a human antibody. Human antibodies can be prepared using various techniques known in the art. In some embodiments, human antibodies are generated from immortalized human B lymphocytes immunized in vitro. In some embodiments, human antibodies are generated from lymphocytes isolated from an immunized individual. In any case, cells that produce an antibody directed against a target antigen can be generated and isolated. In some embodiments, a human antibody is selected from a phage library, where that phage library expresses human antibodies. Alternatively, phage display technology may be used to produce human antibodies and antibody fragments in vitro , from immunoglobulin variable region gene repertoires from unimmunized donors. Techniques for the generation and use of antibody phage libraries are well known in the art. Once antibodies are identified, affinity maturation strategies known in the art, including but not limited to, chain shuffling and site-directed mutagenesis, may be employed to generate higher affinity human antibodies. In some embodiments, human antibodies are produced in transgenic mice that contain human immunoglobulin loci. Upon immunization these mice are capable of producing the full repertoire of human antibodies in the absence of endogenous immunoglobulin production.
[00118] In some embodiments, a C3 -binding agent is an antibody fragment. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv, single chain antibody molecules, scFv, disulfide-linked scFv (dsscFv), nanobodies, diabodies, tribodies, tetrabodies, minibodies, dual variable domain antibodies (DVD), single variable domain antibodies ( e.g ., camelid antibodies), and multispecific antibodies comprising antibody fragments.
[00119] In some embodiments, a C3-binding agent is an scFv antibody. ScFv antibodies are molecules that comprise a variable heavy chain region and a variable light chain region linked to form a single polypeptide. In some embodiments, a scFv comprises a disulfide bond formed between the heavy chain variable region and the light chain variable region. In some embodiments, a scFv antibody comprises a disulfide bond that increases stability of the scFv molecule. In some embodiments, a scFv antibody comprises a disulfide bond that increases thermostability of the scFv molecule. ScFv antibodies can be produced using recombinant technologies known in the art.
[00120] Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody. The antibody fragments described herein can be produced using recombinant technologies known in the art (e.g., E.coli or phage expression).
[00121] In some embodiments, a C3-binding agent is a bispecific antibody. Bispecific antibodies are capable of recognizing and binding at least two different antigens or epitopes. The different epitopes can either be within the same molecule (e.g, two epitopes on C3) or on different molecules (e.g, one epitope on C3 and one epitope on a different target). In some embodiments, a bispecific antibody has enhanced potency as compared to an individual antibody or to a combination of more than one antibody. In some embodiments, a bispecific antibody has reduced toxicity as compared to an individual antibody or to a combination of more than one antibody. It is known to those of skill in the art that any therapeutic agent may have unique pharmacokinetics (PK) (e.g, circulating half-life). In some embodiments, a bispecific antibody has the ability to synchronize the PK of two active binding agents wherein the two individual binding agents have different PK profiles. In some embodiments, a bispecific antibody has the ability to concentrate the actions of two agents in a common area (e.g, tissue) in a subject. In some embodiments, a bispecific antibody has the ability to concentrate the actions of two agents to a common target ( e.g ., a specific cell type). In some embodiments, a bispecific antibody has the ability to target the actions of two agents to more than one biological pathway or function. In some embodiments, a bispecific antibody has the ability to target two different cells and bring them closer together. [00122] In some embodiments, a bispecific antibody has decreased toxicity and/or side effects. In some embodiments, a bispecific antibody has decreased toxicity and/or side effects as compared to a mixture of the two individual antibodies or the antibodies as single agents. In some embodiments, a bispecific antibody has an increased therapeutic index. In some embodiments, a bispecific antibody has an increased therapeutic index as compared to a mixture of the two individual antibodies or the antibodies as single agents.
[00123] Several techniques for making bispecific antibodies are known by those skilled in the art. In some embodiments, the bispecific antibodies comprise heavy chain constant regions with modifications in the amino acids that are part of the interface between the two heavy chains. These modifications are made to enhance heterodimer formation and generally reduce or eliminate homodimer formation. In some embodiments, the bispecific antibodies are generated using a knobs-into-holes (KIH) strategy. In some embodiments, the bispecific antibodies comprise variant hinge regions incapable of forming disulfide linkages between identical heavy chains (e.g., reduce homodimer formation). In some embodiments, the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered electrostatic interactions. In some embodiments, the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered hydrophobic/hydrophilic interactions. [00124] In some embodiments, a bispecific antibody is an intact/full length antibody.
In some embodiments, a bispecific antibody comprises antibody fragments comprising two antigen-binding sites.
[00125] C3-binding agents with more than two valencies are also contemplated. In some embodiments, trispecific or tetraspecific antibodies are generated. [00126] In some embodiments, a C3-binding agent is an antibody that binds C3. In some embodiments, an anti-C3 antibody binds human C3. In some embodiments, an anti-C3 antibody binds cyno C3. In some embodiments, an anti-C3 antibody binds human C3 and cyno C3. In some embodiments, an anti-C3 antibody does not bind rat C3. In some embodiments, an anti-C3 antibody binds a C3 epitope. In some embodiments, an anti-C3 antibody does not bind C3a, C3b, C3c, C3d, or iC3b. In some embodiments, an anti-C3 antibody does not bind C3a, C3b, C3c, C3d, and iC3b. In some embodiments, an anti-C3 antibody does not bind C3a. In some embodiments, an anti-C3 antibody does not bind C3c. In some embodiments, an anti-C3 antibody does not bind C3d. In some embodiments, an anti-C3 antibody does not bind iC3b. In some embodiments, an anti-C3 antibody does not bind C3b. In some embodiments, an anti- C3 antibody does not bind C3a, C3b, C3c, C3d, or iC3b at a detectable level. In some embodiments, an anti-C3 antibody binds C3 with an affinity that is at least 20-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b. In some embodiments, an anti-C3 antibody binds C3 with an affinity that is at least 50-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b. In some embodiments, an anti-C3 antibody binds C3 with an affinity that is at least 100-fold greater than the antibody’s affinity to C3a, C3c, C3d, iC3b or C3b. In some embodiments, an anti-C3 antibody binds C3 and does not bind C3b at a detectable level. [00127] In some embodiments, a C3-binding agent is a variant of an anti-C3 antibody described herein. A variant of an anti-C3 antibody described herein must still bind C3.
In some embodiments, a variant of an anti-C3 antibody described herein comprises one to thirty amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to twenty-five amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to twenty amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to fifteen amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to ten amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to five amino acid substitutions. In some embodiments, the variant of the anti-C3 antibody comprises one to three amino acid substitutions. In some embodiments, the amino acid substitution(s) is in a CDR of the antibody. In some embodiments, the amino acid substitution(s) is not in a CDR of the antibody. In some embodiments, the amino acid substitution(s) is in a framework region of the antibody.
In some embodiments, the amino acid substitution(s) is a conservative amino acid substitution.
[00128] CDRs of an antibody are defined by a variety of methods/sy stems by those skilled in the art. These systems and/or definitions have been developed and refined over a number of years and include Rabat, Chothia, IMGT, AbM, and Contact. The Rabat definition is based on sequence variability and generally is the most commonly used. The Chothia definition is based on the location of the structural loop regions.
The IMGT system is based on sequence variability and location within the structure of the variable domain. The AbM definition is a compromise between Rabat and Chothia. The Contact definition is based on analyses of the available antibody crystal structures. An Exemplary system is a combination of Rabat and Chothia. Software programs ( e.g ., abYsis) are available and known to those of skill in the art for analysis of antibody sequences and determination of CDRs.
[00129] The specific CDR sequences defined herein are generally based on a combination of Rabat and Chothia definitions (Exemplary system). However, it will be understood that reference to a heavy chain CDR or CDRs and/or a light chain CDR or CDRs of a specific antibody will encompass all CDR definitions as known to those of skill in the art.
[00130] In some embodiments, a C3-binding agent comprises one, two, three, four, five, and/or six CDRs of any one of the antibodies described herein. In some embodiments, an anti-C3 antibody comprises one, two, three, four, five, and/or six CDRs of any one of the antibodies described herein. Anti-C3 antibodies have been described in U.S. Patent Application No. 2019/0322730 including antibodies 38G10, Hz38G10, Hz38G10(G56A), Hz38G10(G56T), Hz38G10(N55E), Hz38G10(N55Q), 3D8, 3G8, 1502, 27A8, 28C3, 38F5, 62B11, 62F2, and 63 A3 described herein.
[00131] In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 1 A, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 1A. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table IB, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table IB. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 1C, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 1C.
[00132] In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 2, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 2. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 3, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 3. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 4, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 4. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 5, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 5. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 6, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 6. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 7, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 7. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 8, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 8. In some embodiments, a C3 -binding agent comprises (i) a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 9, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 9. In some embodiments, a C3-binding agent comprises (i) a heavy chain variable region comprising a heavy chain CDR1, a heavy chain CDR2, and/or a heavy chain CDR3 from Table 10, and/or (ii) a light chain variable region comprising a light chain CDR1, a light chain CDR2, and/or a light chain CDR3 from Table 10.
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Table 1A: Antibody 38G10 Sequences
Figure imgf000075_0001
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Table IB: Antibody Hz38G10 and Variant Hz38G10(G56A) Sequences
Figure imgf000076_0001
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Table 1C: Antibody Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q) Sequences
Figure imgf000077_0001
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Table 1C (Continued): Antibody Hz38G10(G56T), Hz38G10(N55E) and Hz38G10(N55Q) Sequences
Figure imgf000078_0001
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Table 2: Antibody 3D8 Sequences
Figure imgf000079_0001
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Table 3: Antibody 3G8 Sequences
Figure imgf000080_0001
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Table 4: Antibody 15C12 Sequences
Figure imgf000081_0001
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Table 5: Antibody 27 A8 Sequences
Figure imgf000082_0001
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Table 6: Antibody 28C3 Sequences
Figure imgf000083_0001
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Table 7: Antibody 38F5 Sequences
Figure imgf000084_0001
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Table 8: Antibody 62B11 Sequences
Figure imgf000085_0001
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Table 9: Antibody 62F2 Sequences
Figure imgf000086_0001
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Table 10: Antibody 63A3 Sequences
Figure imgf000087_0001
[00134] In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from an antibody described herein. In some embodiments, a C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from an antibody described herein. In some embodiments, a C3-binding agent comprises a humanized version or humanized variant of an antibody described herein. In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from antibody 38G10 or a humanized version thereof. In some embodiments, a C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and/or a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10 or variants thereof (e.g., HzG38G10(G56A), HzG38G10(G56T), HzG38G10(N55E), and HzG38G10(N55Q)). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody 38G10. In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10. In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(G56A). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(G56T). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(N55E). In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, CDR3 from antibody Hz38G10(N55Q). [00135] In some embodiments, a C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 using any of the CDR definitions exemplified in Table 1A, Table IB, Table 1C, or Tables 2-10.
[00136] In some embodiments, a C3-binding agent comprises one or more of the following CDRs: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHN GGTT YN QNF T G (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3- binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprises a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprises a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3- binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
[00137] In certain embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence YPHNGG (SEQ ID NO: 16), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (c) GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTT (SEQ ID NO: 17), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (d) DFYMD (SEQ ID NO: 18), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO:l 1), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); or (e) TDFYMD (SEQ ID NO: 19), a heavy chain CDR2 comprising the amino acid sequence WIGYIYPHNGGTT (SEQ ID NO:20), a heavy chain CDR3 comprising the amino acid sequence ARRGGFDFD (SEQ ID NO:21), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence DTYVSWY (SEQ ID NO:22), a light chain CDR2 comprising the amino acid sequence LLIYGASNRY (SEQ ID NO:23), and a light chain CDR3 comprising the amino acid sequence GQSHSYPL (SEQ ID NO:24). In certain embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence YPHNGG (SEQ ID NO: 16), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (c) GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTT (SEQ ID NO: 17), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (d) DFYMD (SEQ ID NO: 18), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO:l 1), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); or (e) TDFYMD (SEQ ID NO: 19), a heavy chain CDR2 comprising the amino acid sequence WMGYIYPHNGGTT (SEQ ID NO:26), a heavy chain CDR3 comprising the amino acid sequence ARRGGFDFD (SEQ ID NO:21), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence DTYVSWY (SEQ ID NO:22), a light chain CDR2 comprising the amino acid sequence LLIYGASNRY (SEQ ID NO:23), and a light chain CDR3 comprising the amino acid sequence GQSHSYPL (SEQ ID NO:24). In certain embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO: 9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO:l 1), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (b) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDF (SEQ ID NO: 15), a heavy chain CDR2 comprising the amino acid sequence YPHNAG (SEQ ID NO:28), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (c) GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTT (SEQ ID NO:29), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); (d) DFYMD (SEQ ID NO: 18), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14); or (e) TDFYMD (SEQ ID NO: 19), a heavy chain CDR2 comprising the amino acid sequence WMGYIYPHNAGTT (SEQ ID NO:30), a heavy chain CDR3 comprising the amino acid sequence ARRGGFDFD (SEQ ID NO:21), and a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence DTYVSWY (SEQ ID NO:22), a light chain CDR2 comprising the amino acid sequence LLIYGASNRY (SEQ ID NO:23), and a light chain CDR3 comprising the amino acid sequence GQSHSYPL (SEQ ID NO:24).
[00138] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3- binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3- binding agent comprises a light chain variable region comprising light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
[00139] In some embodiments, a C3-binding agent is a variant of an agent described herein. A C3-binding agent can comprise a heavy chain variable region comprising a heavy chain CDR1 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR1 described herein; a heavy chain CDR2 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR2 described herein; a heavy chain CDR3 with 1, 2, 3, or 4 amino acid substitutions of a heavy chain CDR3 described herein; a light chain CDR1 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR1 described herein; a light chain CDR2 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR2 described herein; and a light chain CDR3 with 1, 2, 3, or 4 amino acid substitutions of a light chain CDR3 described herein. For example, in some embodiments, a C3 -binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9) or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10) or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14), or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions.
[00140] In some embodiments, a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification reduces deamidation. In some embodiments, a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to reduce deamidation within the CDR sequence. Deamidation is a chemical reaction in which an amide functional group in the side chain of the amino acids asparagine (Asn or N) or glutamine (Gin or Q) is removed or converted to another functional group. Generally, asparagine is converted to aspartic acid or isoaspartic acid and glutamine is converted to glutamic acid or polyglutamic acid. In some situations, deamidation may change the structure, function, and/or stability of a polypeptide, potentially resulting in decreased biological activity.
[00141] In some embodiments, a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification reduces isomerization. In some embodiments, a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to reduce isomerization. Isomerization is a chemical process by which a compound is transformed into any of its isomeric forms, i.e., forms with the same chemical composition but with different structure or configuration and, potentially with different physical and chemical properties. Studies have shown that aspartate (Asp or D) isomerization within a CDR can impact antibody binding and/or stability.
[00142] In some embodiments, a C3-binding agent comprises a heavy chain variable region and/or a light chain variable region that comprises a modification within the amino acid sequence wherein the modification eliminates a glycosylation site. In some embodiments, a C3-binding agent comprises one or more heavy chain variable region CDRs or light chain variable region CDRs that have been modified to eliminate a glycosylation site. The consensus glycosylation site for N-linked glycans is N-X-S/T, wherein X can be any amino acid except proline. Generally, a glycosylation site within a variable region and/or within a CDR will impact antibody structure, binding, and/or stability.
[00143] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprise a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). [00144] In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and/or (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11). In some embodiments, a C3-binding agent comprise a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments, a C3-binding agent comprises: (a) a heavy chain variable region comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). [00145] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, or SEQ ID NO:224; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32 or SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 31. In some embodiments, a C3- binding agent comprises a light chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:33. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:34. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:222. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:223. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:224. In some embodiments, a C3-binding agent comprises a light chain variable region having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO:35. [00146] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31 and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 31 and a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31 and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 31 and a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 31 and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:31 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:32.
[00147] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:34 and a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
[00148] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:222; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
[00149] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:223; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
[00150] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:224; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
[00151] In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:32. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:32. [00152] In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:33 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:34 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
[00153] In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:222, SEQ ID NO:223 or SEQ ID NO:224. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:222 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:223 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, a C3 -binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:224 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35.
[00154] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:77; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:78. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:78. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:77 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:78.
[00155] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:96. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:96. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:95; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:96. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:95. In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:96. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:95 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:96.
[00156] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 113; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 113; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:l 13; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 113.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 114. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 113 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 114.
[00157] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 131; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 131.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 132. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 131 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 132.
[00158] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 149; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 149.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 149 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 150.
[00159] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3 -binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 167; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 167.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 167 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 168.
[00160] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 185; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 185.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 185 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 186.
[00161] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:203; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:203.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:204. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:203 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:204.
[00162] In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3-binding agent comprises a heavy chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:220; and/or a light chain variable region having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3-binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:220.
In some embodiments, a C3-binding agent comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:221. In some embodiments, a C3- binding agent comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:220 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 221.
[00163] In some embodiments, a C3-binding agent described herein comprises an antibody in which at least one or more of the constant regions has been modified or deleted. In some embodiments, the antibodies comprise at least one modification in the hinge region of the heavy chain. In some embodiments, the antibodies comprise modifications to one or more of the three heavy chain constant regions (CHI, CH2 or CH3) and/or to the light chain constant region (CL). In some embodiments, the heavy chain constant region of the modified antibodies comprises at least one human constant region. In some embodiments, the heavy chain constant region of the modified antibodies comprises more than one human constant region. In some embodiments, modifications to the constant region comprise additions, deletions, or substitutions of one or more amino acids in one or more regions. In some embodiments, one or more regions are partially or entirely deleted from the constant regions of the modified antibodies. In some embodiments, the entire CH2 domain has been removed from an antibody (ΔCH2 constructs). In some embodiments, a deleted constant region is replaced by a short amino acid spacer that provides some of the molecular flexibility typically imparted by the absent constant region. In some embodiments, a modified antibody comprises a CH3 domain directly fused to the hinge region of the antibody. In some embodiments, a modified antibody comprises a peptide spacer inserted between the hinge region and modified CH2 and/or CH3 domains.
[00164] It is known in the art that the constant region(s) of an antibody mediates several effector functions and these effector functions can vary depending on the isotype of the antibody. For example, binding of the Cl component of complement to the Fc region of IgG or IgM antibodies (bound to antigen) activates the complement system. Activation of complement is important in the opsonization and lysis of cell pathogens. The activation of complement also stimulates the inflammatory response and can be involved in autoimmune hypersensitivity. In addition, the Fc region of an antibody can bind a cell expressing a Fc receptor (FcR). There are a number of Fc receptors which are specific for different classes of antibody, including IgG (gamma receptors), IgE (epsilon receptors), IgA (alpha receptors) and IgM (mu receptors). Binding of antibody to Fc receptors on cell surfaces triggers a number of important and diverse biological responses including engulfment and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target cells by killer cells (called antibody-dependent cell cytotoxicity or ADCC), release of inflammatory mediators, placental transfer, and control of immunoglobulin production.
[00165] In some embodiments, an antibody comprises a variant Fc region. The amino acid sequences of the Fc region of human IgGl, IgG2, IgG3, and IgG4 are known to those of ordinary skill in the art ( e.g ., a representative human IgGl is SEQ ID NO:42). In some cases, Fc regions with amino acid variations have been identified in native antibodies. In some embodiments, a variant Fc region is engineered with substitutions at specific amino acid positions as compared to a native Fc region ( e.g ., SEQ ID NOs:43-46).
[00166] In some embodiments, the modified antibodies provide for altered effector functions that, in turn, affect the biological profile of the antibody. For example, in some embodiments, the deletion or inactivation (through point mutations or other means) of a constant region reduces Fc receptor binding of the modified antibody as it circulates. In some embodiments, the constant region modifications increase the serum half-life of the antibody. In some embodiments, the constant region modifications reduce the serum half-life of the antibody. In some embodiments, the constant region modifications decrease or remove ADCC and/or complement dependent cytotoxicity (CDC) of the antibody. In some embodiments, specific amino acid substitutions in a human IgGl Fc region with corresponding IgG2 or IgG4 residues reduce effector functions (e.g., ADCC and CDC) in the modified antibody. In some embodiments, an antibody does not have one or more effector functions. In some embodiments, the antibody has no ADCC activity and/or no CDC activity. In some embodiments, the antibody does not bind an Fc receptor and/or complement factors. In some embodiments, the antibody has no effector function(s) (e.g, “effectorless” antibodies). In some embodiments, the constant region modifications increase or enhance ADCC and/or CDC of the antibody. In some embodiments, the constant region is modified to eliminate disulfide linkages or oligosaccharide moieties. In some embodiments, the constant region is modified to add/substitute one or more amino acids to provide one or more cytotoxin, oligosaccharide, or carbohydrate attachment sites.
[00167] In certain embodiments, a C3-binding agent comprises (a) a heavy chain comprising a heavy chain CDR1, a heavy chain CDR2, and a heavy chain CDR3 using any one of the CDR definitions as shown in Tables 1 A- 1C, and (b) a light chain comprising a light chain CDR1, a light chain CDR2, and a light chain CDR3 using any of the CDR definitions as shown in Tables 1 A-1C, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and a light chain comprising a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14), wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO: 10, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:25, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:27, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:48, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:52, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO:39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41. In certain embodiments, a C3-binding agent comprises a heavy chain comprising a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 9, a heavy chain CDR2 comprising the amino acid sequence SEQ ID NO:56, a heavy chain CDR3 comprising the amino acid sequence SEQ ID NO: 11, and a light chain comprising a light chain CDR1 comprising the amino acid sequence SEQ ID NO: 12, a light chain CDR2 comprising the amino acid sequence SEQ ID NO: 13, and a light chain CDR3 comprising the amino acid sequence SEQ ID NO: 14, wherein the heavy chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:37 or SEQ ID NO: 39, and wherein the light chain comprises at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or 100% sequence identity to the sequence of SEQ ID NO:41.
[00168] In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39 and/or a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39. In some embodiments, a C3-binding agent is an antibody that comprises a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:37 and a light chain of SEQ ID NO:41. In some embodiments, a C3-binding agent is an antibody that comprises a heavy chain of SEQ ID NO:39 and a light chain of SEQ ID NO:41.
[00169] Modifications to the constant region of antibodies described herein may be made using well known biochemical or molecular engineering techniques. In some embodiments, antibody variants are prepared by introducing appropriate nucleotide changes into the encoding DNA, and/or by synthesis of the desired antibody or polypeptide. Using these antibody variants it may be possible to disrupt the activity or effector function provided by a specific sequence or region while substantially maintaining the structure, binding activity, and other desired characteristics of the modified antibody.
[00170] The present disclosure further embraces additional variants and equivalents that are substantially homologous to the recombinant, monoclonal, chimeric, humanized, and human antibodies, or antibody fragments thereof, described herein. In some embodiments, it is desirable to improve the binding affinity of the antibody. In some embodiments, it is desirable to modulate biological properties of the antibody, including but not limited to, specificity, thermostability, expression level, effector function(s), glycosylation, immunogenicity, or solubility. Those skilled in the art will appreciate that amino acid changes may alter post-translational processes of an antibody, such as changing the number or position of glycosylation sites or altering membrane anchoring characteristics.
[00171] Variations may be a substitution, deletion, or insertion of one or more nucleotides encoding the antibody or polypeptide that results in a change in the amino acid sequence as compared with the native antibody or polypeptide sequence. In some embodiments, amino acid substitutions are the result of replacing one amino acid with another amino acid having similar structural and/or chemical properties, such as the replacement of a leucine with a serine, e.g., conservative amino acid substitutions. Insertions or deletions may optionally be in the range of about 1 to 5 amino acids. In some embodiments, the substitution, deletion, or insertion includes less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the parent molecule. In some embodiments, variations in the amino acid sequence that are biologically useful and/or relevant are determined by systematically making insertions, deletions, or substitutions in the sequence and testing the resulting variant proteins for activity as compared to the parental antibody.
[00172] In some embodiments, variants may include addition of amino acid residues at the amino- and/or carboxyl-terminal end of the antibody or polypeptide. The length of additional amino acids residues may range from one residue to a hundred or more residues. In some embodiments, a variant comprises an N-terminal methionyl residue. In some embodiments, a variant does not comprise an N-terminal methionyl residue. In some embodiments, the variant comprises an additional polypeptide/protein, /. e. , a fusion protein. In some embodiments, a variant is engineered to be detectable and may comprise a detectable label and/or protein (e.g, an enzyme).
[00173] In some embodiments, a cysteine residue not involved in maintaining the proper conformation of an antibody may be substituted or deleted to modulate the antibody’s characteristics, for example, to improve oxidative stability and/or prevent aberrant disulfide crosslinking. Conversely, in some embodiments, one or more cysteine residues may be added to create disulfide bond(s) to improve stability.
[00174] In some embodiments, an antibody of the present disclosure is “deimmunized”. The deimmunization of antibodies generally consists of introducing specific amino acid mutations (e.g., substitutions, deletions, additions) that result in removal of T-cell epitopes without significantly reducing the binding affinity or other desired activities of the antibody. [00175] The variant antibodies or polypeptides described herein may be generated using methods known in the art, including but not limited to, site-directed mutagenesis, alanine scanning mutagenesis, and PCR mutagenesis.
[00176] In some embodiments, C3-binding agents described herein are chemically modified. In some embodiments, the C3 -binding agents are anti-C3 antibodies that have been chemically modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, and/or linkage to a cellular ligand or other protein. Any of numerous chemical modifications may be carried out by known techniques.
[00177] In some embodiments, a C3-binding agent is attached, either directly or indirectly, to a half-life extending moiety including, but not limited to, a Fc region, a CH3 domain of an immunoglobulin, polyethylene glycol (PEG), a PEG mimetic, XTEN®, serum albumin, polysialic acid, N-(2-hydroxypropyl)methacrylamide, or dextran. In some embodiments, an anti-C3 antibody described herein is attached, either directly or indirectly, to a half-life extending moiety including, but not limited to, polyethylene glycol (PEG), a PEG mimetic, XTEN®, serum albumin, polysialic acid, N-(2-hydroxypropyl)methacrylamide, or dextran.
[00178] The present disclosure encompasses C3-binding agents built upon nonimmunoglobulin backbones, wherein the agents bind to the same epitope or essentially the same epitope as an anti-C3 antibody disclosed herein. In some embodiments, a non- immunoglobulin-based binding agent is an agent that competes with an anti-C3 antibody described herein in a competitive binding assay. In some embodiments, alternative C3-binding agents comprise a scaffold protein. Generally, scaffold proteins can be assigned to one of three groups based on the architecture of their backbone (1) scaffolds consisting of a-helices; (2) small scaffolds with few secondary structures or an irregular architecture of a-helices and b-sheets; and (3) scaffolds consisting of predominantly b-sheets. Scaffold proteins include, but are not limited to, anticalins, which are based upon the lipocalin scaffold; adnectins, which are based on the 10th domain of human fibronectin type 3; affibodies, which are based on the B-domain in the Ig-binding region of Staphylococcus aureus protein A; darpins, which are based on ankyrin repeat domain proteins; fynomers, which are based on the SH3 domain of the human Fyn protein kinase; affitins, which are based on Sac7d from Sulfolobus acidocaldarius ; affilins, which are based on human g-B-crystallin or human ubiquitin; avimers, which are based on the A-domains of membrane receptor proteins; knottins (cysteine knot miniproteins), which are based upon a stable 30-amino acid anti -parallel b-strand protein fold; and Kunitz domain inhibitor scaffolds, which are based upon a structure that contains three disulfide bonds and three loops. In some embodiments, a C3-binding agent comprises an engineered scaffold protein comprising a heavy chain CDR1, CDR2, and CDR3 and a light chain CDR1, CDR2, and CDR3 shown in Tables 1A-1C or Tables 2-10. In some embodiments, a C3-binding agent comprises an engineered scaffold protein comprising a heavy chain CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain CDR2 comprising the amino acid sequence YIYPHNGGTTYNQNFTG (SEQ ID NO: 10),
YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), a light chain CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
[00179] Generally speaking, antigen-antibody interactions are non-covalent and reversible, formed by a combination of hydrogen bonds, hydrophobic interactions, electrostatic and van der Waals forces. When describing the strength of an antigen- antibody complex, the terms affinity and/or avidity are commonly used mentioned. The binding of an antibody to its antigen is a reversible process, and the affinity of the binding is typically reported as an equilibrium dissociation constant (KD). KD is the ratio of an antibody dissociation rate (k0ff) (how quickly it dissociates from its antigen) to the antibody association rate (k0n) (how quickly it binds to its antigen). In some embodiments, KD values are determined by measuring the k0n and k0ff rates of a specific antibody/antigen interaction and then using a ratio of these values to calculate the KD value. KD values may be used to evaluate and rank order the strength of individual antibody/antigen interactions. The lower the KD of an antibody, the higher the affinity of the antibody for its target. In some embodiments, affinity is measured using SPR technology in a Biacore system. Avidity gives a measure of the overall strength of an antibody-antigen complex. It is dependent on three major parameters: (i) affinity of the antibody for the target, (ii) valency of both the antibody and antigen, and (iii) structural arrangement of the parts that interact.
[00180] In some embodiments, a C3 -binding agent binds C3 with a dissociation constant (KD) of about 1 mM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, about 0.1 nM or less, 50 pM or less, 10 pM or less, or 1 pM or less. In some embodiments, a C3 -binding agent binds C3 with a KD of about 20 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 10 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 1 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 0.5 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 0.1 nM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 50 pM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 25 pM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 10 pM or less. In some embodiments, a C3-binding agent binds C3 with a KD of about 1 pM or less. In some embodiments, the dissociation constant of the binding agent for C3 is the dissociation constant determined using a C3 protein immobilized on a Biacore chip and the binding agent flowed over the chip. In some embodiments, the dissociation constant of the binding agent for C3 is the dissociation constant determined using the binding agent captured by an anti-human IgG antibody on a Biacore chip and soluble C3 flowed over the chip.
[00181] In some embodiments, a C3 -binding agent binds C3 with a half maximal effective concentration (EC50) of about 1 pM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 nM or less. In some embodiments, a C3 -binding agent binds to human C3 with an EC50 of about 1 pM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 nM or less. In some embodiments, a C3-binding agent binds cyno C3 and/or human C3 with an EC50 of about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less or about 0.1 nM or less.
[00182] The C3 -binding agents described herein can be produced by any suitable method known in the art. Such methods range from direct protein synthesis methods to constructing a DNA sequence encoding polypeptide sequences and expressing those sequences in a suitable host. In some embodiments, a DNA sequence is constructed using recombinant technology by isolating or synthesizing a DNA sequence encoding a wild-type protein of interest. Optionally, the sequence can be mutagenized by site- specific mutagenesis to provide functional variants thereof. In some embodiments, a DNA sequence encoding a polypeptide of interest is constructed by chemical synthesis using an oligonucleotide synthesizer. Oligonucleotides can be designed based on the amino acid sequence of the desired polypeptide and selecting those codons that are favored in the host cell in which the recombinant polypeptide of interest will be produced. Standard methods can be applied to synthesize a polynucleotide sequence encoding an isolated polypeptide of interest. For example, a complete amino acid sequence can be used to construct a back-translated gene. Further, a DNA oligomer containing a nucleotide sequence coding for the particular isolated polypeptide can be synthesized. For example, several small oligonucleotides coding for portions of the desired polypeptide can be synthesized and then ligated. The individual oligonucleotides typically contain 5' or 3' overhangs for complementary assembly. [00183] Once assembled (by synthesis, site-directed mutagenesis, or another method), the polynucleotide sequences encoding a particular polypeptide of interest can be inserted into an expression vector and operatively linked to an expression control sequence appropriate for expression of the protein in a desired host. Proper assembly can be confirmed by nucleotide sequencing, restriction enzyme mapping, and/or expression of a biologically active polypeptide in a suitable host. As is well-known in the art, in order to obtain high expression levels of a transfected gene in a host, the gene must be operatively linked to transcriptional and translational expression control sequences that are functional in the chosen expression host. [00184] In some embodiments, recombinant expression vectors are used to amplify and express DNA encoding antibodies, or fragments thereof, against human C3. For example, recombinant expression vectors can be replicable DNA constructs which have synthetic or cDNA-derived DNA fragments encoding a polypeptide chain of a C3- binding agent, such as an anti-C3 antibody, or antigen-binding fragment thereof, operatively linked to suitable transcriptional and/or translational regulatory elements derived from mammalian, microbial, viral or insect genes. A transcriptional unit generally comprises an assembly of (1) a genetic element or elements having a regulatory role in gene expression, for example, transcriptional promoters or enhancers, (2) a structural or coding sequence which is transcribed into mRNA and translated into protein, and (3) appropriate transcription and translation initiation and termination sequences. Regulatory elements can include an operator sequence to control transcription. The ability to replicate in a host, usually conferred by an origin of replication, and a selection gene to facilitate recognition of transformants can additionally be incorporated. DNA regions are “operatively linked” when they are functionally related to each other. For example, DNA for a signal peptide (secretory leader) is operatively linked to DNA for a polypeptide if it is expressed as a precursor which participates in the secretion of the polypeptide; a promoter is operatively linked to a coding sequence if it controls the transcription of the sequence; or a ribosome binding site is operatively linked to a coding sequence if it is positioned so as to permit translation. In some embodiments, structural elements intended for use in yeast expression systems include a leader sequence enabling extracellular secretion of translated protein by a host cell. In some embodiments, in situations where recombinant protein is expressed without a leader or transport sequence, a polypeptide may include an N-terminal methionine residue. This residue can optionally be subsequently cleaved from the expressed recombinant protein to provide a final product. [00185] The choice of an expression control sequence and an expression vector generally depends upon the choice of host. A wide variety of expression host/vector combinations can be employed. Useful expression vectors for eukaryotic hosts include, for example, vectors comprising expression control sequences from SV40, bovine papilloma virus, adenovirus, and cytomegalovirus. Useful expression vectors for bacterial hosts include known bacterial plasmids, such as plasmids from E. coli , including pCRl, pBR322, pMB9 and their derivatives, and wider host range plasmids, such as Ml 3 and other filamentous single-stranded DNA phages.
[00186] The C3-binding agents of the present disclosure can be expressed from one or more vectors. For example, in some embodiments, a heavy chain polypeptide is expressed by one vector and a light chain polypeptide is expressed by a second vector.
In some embodiments, a heavy chain polypeptide and a light chain polypeptide are expressed by one vector.
[00187] Suitable host cells for expression of a C3-binding agent or a C3 protein or fragment thereof to use as an antigen or immunogen include prokaryotes, yeast cells, insect cells, or higher eukaryotic cells under the control of appropriate promoters. Prokaryotes include gram-negative or gram-positive organisms, for example E. coli or Bacillus. Higher eukaryotic cells include established cell lines of mammalian origin as described herein. Cell-free translation systems may also be employed. Appropriate cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cellular hosts, as well as methods of protein production, including antibody production are well known in the art.
[00188] Various mammalian culture systems may be used to express recombinant polypeptides. Expression of recombinant proteins in mammalian cells may be desirable because these proteins are generally correctly folded, appropriately modified, and biologically functional. Examples of suitable mammalian host cell lines include, but are not limited to, COS-7 (monkey kidney-derived), L-929 (murine fibroblast-derived),
Cl 27 (murine mammary tumor-derived), 3T3 (murine fibroblast-derived), CHO (Chinese hamster ovary-derived), HeLa (human cervical cancer-derived), BHK (hamster kidney fibroblast-derived), HEK-293 (human embryonic kidney-derived) cell lines and variants thereof. Mammalian expression vectors can comprise non- transcribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, and other 5' or 3' flanking non-transcribed sequences, and 5' or 3' non-translated sequences, such as necessary ribosome binding sites, a polyadenylation site, splice donor and acceptor sites, and transcriptional termination sequences.
[00189] Expression of recombinant proteins in insect cell culture systems ( e.g ., baculovirus) also offers a robust method for producing correctly folded and biologically functional proteins. Baculovirus systems for production of heterologous proteins in insect cells are well-known to those of skill in the art.
[00190] Thus, the present disclosure provides cells comprising the C3-binding agents described herein. In some embodiments, the cells produce the C3-binding agents described herein. In some embodiments, the cells produce an antibody. In some embodiments, the cells produce an antibody that binds human C3. In some embodiments, the cells produce an antibody that binds cyno C3. In some embodiments, the cells produce an antibody that binds human C3 and cyno C3. In some embodiments, the cells produce an antibody designated 38G10. In some embodiments, the cells produce a humanized version of antibody 38G10, referred to as Hz38G10. In some embodiments, the cells produce a variant of Hz38G10. In some embodiments, the cells produce antibody Hz38G10(G56A). In some embodiments, the cell is a prokaryotic cell. In some embodiments, the cell is an eukaryotic cell. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell is a hybridoma cell.
[00191] Proteins produced by a host cell can be purified according to any suitable method. Standard methods include chromatography (e.g., ion exchange, affinity, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for protein purification. Affinity tags such as hexa-histidine (SEQ ID NO:47), maltose binding domain, influenza coat sequence, and glutathione-S- transferase can be attached to the protein to allow easy purification by passage over an appropriate affinity column. Affinity chromatography used for purifying immunoglobulins can include Protein A, Protein G, and Protein L chromatography. Isolated proteins can be physically characterized using such techniques as proteolysis, size exclusion chromatography (SEC), mass spectrometry (MS), nuclear magnetic resonance (NMR), isoelectric focusing (IEF), high performance liquid chromatography (HPLC), and x-ray crystallography. The purity of isolated proteins can be determined using techniques known to those of skill in the art, including but not limited to, SDS- PAGE, SEC, capillary gel electrophoresis, IEF, and capillary isoelectric focusing (cIEF).
[00192] In some embodiments, supernatants from expression systems which secrete recombinant protein into culture media are first concentrated using a commercially available protein concentration filter, for example, an Amicon® or Millipore Pellicon® ultrafiltration unit. Following the concentration step, the concentrate can be applied to a suitable purification matrix. In some embodiments, an anion exchange resin is employed, for example, a matrix or substrate having pendant diethylaminoethyl (DEAE) groups. The matrices can be acrylamide, agarose, dextran, cellulose, or other types commonly employed in protein purification. In some embodiments, a cation exchange step is employed. Suitable cation exchangers include various insoluble matrices comprising sulfopropyl or carboxymethyl groups. In some embodiments, a hydroxyapatite media is employed, including but not limited to, ceramic hydroxyapatite (CHT). In some embodiments, one or more reverse-phase HPLC steps employing hydrophobic RP-HPLC media, e.g, silica gel having pendant methyl or other aliphatic groups, are employed to further purify a recombinant protein. In some embodiments, hydrophobic interaction chromatography (HIC) is used to separate recombinant proteins based on their hydrophobicity. HIC is a useful separation technique for purifying proteins while maintaining biological activity due to the use of conditions and matrices that operate under less denaturing conditions than some other techniques. Some or all of the foregoing purification steps, in various combinations, can be employed to provide a homogeneous recombinant protein.
[00193] Anti-C3 antibodies of the present disclosure may be analyzed for their physical/chemical properties and/or biological activities by various assays known in the art. In some embodiments, an anti-C3 antibody is tested for its ability to bind C3 (e.g, human C3 and/or cyno C3). Binding assays include, but are not limited to, SPR (e.g, Biacore), ELISA, and FACS. In some embodiments, an anti-C3 antibody is tested for its ability to inhibit, reduce, or block complement activity. In some embodiments, an anti-C3 antibody is tested for its ability to inhibit, reduce, or block C3 activity. Assays include, but are not limited to, hemolysis assays and C3a release assays. In addition, antibodies may be evaluated for solubility, stability, thermostability, viscosity, expression levels, expression quality, and/or purification efficiency.
[00194] In some embodiments, monoclonal antibodies generated against C3 are grouped based upon the epitope each individual antibody recognizes, a process known as “epitope binning”. Generally, antibodies are tested in a pairwise combinatorial manner and antibodies that compete with each other are grouped together into bins. For example, in a premix binning assay, a first antibody is immobilized on a surface and a premixed solution of a second antibody and antigen is flowed over the immobilized first antibody. In tandem, the antigen is immobilized on a surface and the two antibodies are flowed over the immobilized antigen and compete to bind. Using these techniques, antibodies that block one another can be identified. A competitive blocking profile is created for each antibody relative to the other antibodies. The blocking results determine which bin each antibody is placed in. High-throughput methods of epitope binning are known in the art and allow for screening and characterization of large numbers of antibodies within a short period of time. Antibodies that bind similar epitopes often share similar functions and/or capabilities. Conversely, antibodies that bind different epitopes may have different functional activities.
[00195] Epitope mapping is the process of identifying the binding site, or epitope on a target protein/antigen where an antibody (or other binding agent) binds. A variety of methods are known in the art for mapping epitopes on target proteins. These methods include mutagenesis, including but not limited to, shotgun mutagenesis, site-directed mutagenesis, and alanine scanning; domain or fragment scanning, peptide scanning ( e.g ., Pepscan technology); display methods (e.g, phage display, microbial display, and ribosome/mRNA display); methods involving proteolysis and mass spectroscopy; and structural determination (e.g, x-ray crystallography and NMR).
[00196] In some embodiments, purified anti-C3 antibodies are characterized by assays including, but not limited to, N-terminal sequencing, amino acid analysis, high pressure liquid chromatography (HPLC), mass spectrometry, ion exchange chromatography, and papain digestion.
[00197] In some embodiments, assays are provided for identifying anti-C3 antibodies that affect C3 activity. In some embodiments, hemolysis assays are used to assess the functional activity of the complement system. Hemolysis assays have been modified over the years to assess the activity of the different complement pathways and/or individual complement components of the cascade. In some embodiments, a hemolysis assay is used to assess activity of the alternative pathway. In some embodiments, a hemolysis assay is used to assess activity of the classical pathway.
[00198] In some embodiments, a C3 -binding agent binds C3 and inhibits C3 activation of the alternative pathway. In some embodiments, a C3-binding agent binds C3 and inhibits activation of the alternative pathway, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3-binding agent inhibits activation of the alternative pathway by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%. In some embodiments, percent inhibition is used to calculate an ICso (half maximal inhibitory concentration) for the C3 -binding agent. In some embodiments, a C3 -binding agent that inhibits activation of the alternative pathway is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
[00199] In some embodiments, a C3 -binding agent described herein binds C3 and inhibits C3 activation of the classical pathway. In some embodiments, a C3-binding agent binds C3 and inhibits activation of the classical pathway, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3-binding agent inhibits activation of the classical pathway by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%. In some embodiments, a C3-binding agent that inhibits activation of the classical pathway is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
[00200] In some embodiments, a C3 -binding agent described herein binds C3 and inhibits C3 activation of the classical and alternative pathways. In some embodiments, a C3 -binding agent binds C3 and inhibits activation of the classical and alternative pathways, wherein the inhibition is evaluated by a hemolysis assay. In certain embodiments, the C3 -binding agent inhibits activation of the classical and alternative pathways by at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, at least 90%, or about 100%. In some embodiments, a C3 -binding agent that inhibits activation of the classical and alternative pathways is antibody 38G10, antibody Hz38G10, antibody Hz38G10(G56A), antibody Hz38G10(G56T), antibody Hz38G10(N55E), antibody Hz38G10(N55Q), antibody 3D8, antibody 3G8, antibody 15C12, antibody 27A8, antibody 28C3, antibody 38F5, antibody 62B11, antibody 62F2, or antibody 63 A3 described herein.
[00201] In some embodiments, an immunochemical assay to determine the presence and/or amount of individual components ( e.g ., C3a) is used to assess the complement cascade. In some embodiments, a C3-binding agent (e.g., an antibody) described herein binds C3 and inhibits the release of C3a.
[00202] The present disclosure also provides conjugates comprising an anti-C3 antibody described herein. In some embodiments, the antibody is attached to a second molecule. In some embodiments, the antibody is conjugated to a cytotoxic agent or moiety. In some embodiments, the antibody is conjugated to a cytotoxic agent to form an ADC (antibody-drug conjugate). In some embodiments, the cytotoxic agent is a chemotherapeutic agent including, but not limited to, methotrexate, adriamycin/doxorubicin, melphalan, mitomycin C, chlorambucil, duocarmycin, daunorubicin, pyrrolobenzodiazepines (PBDs), or other intercalating agents. In some embodiments, the cytotoxic agent is a microtubule inhibitor including, but not limited to, auristatins, maytansinoids (e.g., DM1 and DM4), and tubulysins. In some embodiments, the cytotoxic agent is an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof, including, but not limited to, diphtheria A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. In some embodiments, an antibody is conjugated to one or more small molecule toxins, such as calicheamicins, maytansinoids, trichothenes, and CC1065. A derivative of any one of these toxins may be used as long as the derivative retains the cytotoxic activity of the parent molecule. [00203] Conjugates comprising an anti-C3 antibody described herein may be made using any suitable method known in the art. In some embodiments, conjugates are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3- (2-pyridyidithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HC1), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis(p- azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis-(p- diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as toluene 2,6-diisocyanate), and bis-active fluorine compounds (such as l,5-difluoro-2, 4-dinitrobenzene).
[00204] In some embodiments, an anti-C3 antibody described herein is conjugated to a detectable substance or molecule that allows the antibody to be used for diagnosis and/or detection. A detectable substance can include but is not limited to, enzymes, such as horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and acetylcholinesterase; prosthetic groups, such as biotin and flavine(s); fluorescent materials, such as, umbelliferone, fluorescein, fluorescein isothiocyanate (FITC), rhodamine, tetramethylrhodamine isothiocyanate (TRITC), dichlorotriazinylamine fluorescein, dansyl chloride, cyanine (Cy3), and phycoerythrin; bioluminescent materials, such as luciferase; radioactive materials, such as 212Bi, 14C, 57Co, 51Cr, 67Cu, 18F, 68Ga, 67Ga, 153Gd, 159Gd, 68Ge, ¾, 166Ho, 131I, 125I, 123I, 121I, 115In, 113In, 112In, mIn, 140La, 177LU, 54Mn, "Mo, 32P, 103Pd, 149Pm, 142Pr, 186Re, 188Re, 105Rh, 97Ru, 35 S, 47Sc, 75Se, 153Sm, 113Sn, 117Sn, 85Sr, 99mTc, 201Ti, 133Xe, 90Y, 69Yb, 175Yb, 65Zn; positron emitting metals; and magnetic metal ions. [00205] An anti-C3 antibody described herein can also be conjugated to a second antibody to form an antibody heteroconjugate.
[00206] An anti-C3 antibody as described herein may be attached to a solid support. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene. In some embodiments, immobilized anti-C3 antibodies are used in immunoassays. In some embodiments, immobilized anti-C3 antibodies are used in purification of the target antigen.
III. Polynucleotides
[00207] The disclosure encompasses a polynucleotide composition comprising a polynucleotide or polynucleotides that encode a C3 -binding agent described herein.
The term “a polynucleotide or polynucleotides that encode a C3 -binding agent” encompasses a polynucleotide or polynucleotides which include only coding sequences for a C3 -binding agent as well as a polynucleotide or polynucleotides which include additional coding and/or non-coding sequences. The polynucleotide or polynucleotides of the disclosure can be in the form of RNA or in the form of DNA. DNA includes cDNA, genomic DNA, and synthetic DNA; and can be double-stranded or single- stranded, and if single stranded can be the coding strand or non-coding (anti-sense) strand.
[00208] In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, provided herein is a polynucleotide or polynucleotides encoding a polypeptide or polypeptides comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41.
[00209] In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:36. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:37. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:38. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:39. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:36 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:37 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:38 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:39 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:36 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:37 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:41. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:38 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:40. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:39 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:41.
[00210] In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NOs:77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224. In some embodiments, provided herein is a polynucleotide or polynucleotides encoding a polypeptide or polypeptides comprising more than one amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149,
150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
[00211] In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:77. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 95. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:96. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 113. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:l 14. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 131. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 132. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 149. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 167. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 168. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 185. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:203. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:204. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:220. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:222. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:223. In some embodiments, the polynucleotide comprises a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:224. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:77 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:95 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:96. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 113 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 114. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 131 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 132. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 149 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 167 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 168. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO: 185 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:203 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:204. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:220 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:222 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:223 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide comprises a polynucleotide encoding (i) a polypeptide comprising an amino acid sequence of SEQ ID NO:224 and (ii) a polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:77 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:78. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:95 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:96. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 113 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 114. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 131 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 132. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 149 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 150. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 167 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 168. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO: 185 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO: 186. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:203 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:204. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:220 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:221. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:222 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:223 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35. In some embodiments, the polynucleotide composition comprises (i) a polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:224 and (ii) a polynucleotide encoding polypeptide comprising an amino acid sequence of SEQ ID NO:35.
[00212] In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98%, or 99% identical to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. Also provided is a polynucleotide that comprises a polynucleotide that hybridizes to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs:31-41. In some embodiments, the hybridization is under conditions of high stringency as is known to those skilled in the art.
[00213] In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98%, or 99% identical to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NO s: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224. Also provided is a polynucleotide that comprises a polynucleotide that hybridizes to a polynucleotide encoding an amino acid sequence selected from the group consisting of: SEQ ID NOs: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224. In some embodiments, the hybridization is under conditions of high stringency as is known to those skilled in the art.
[00214] In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence selected from the group consisting of: SEQ ID NOs:36-41. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:36 or SEQ ID NO:37. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:38 or SEQ ID NO:39. In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence of SEQ ID NO:40 or SEQ ID NO:41.
[00215] In some embodiments, a polynucleotide comprises a polynucleotide having a nucleotide sequence that encodes an amino acid sequence selected from the group consisting of: SEQ ID NO s: 77, 78, 95, 96, 113, 114, 131, 132, 149, 150, 167, 168, 185, 186, 203, 204, 220, 221, and 222-224.
[00216] In some embodiments, a polynucleotide comprises the coding sequence for a polypeptide fused in the same reading frame to a polynucleotide which aids in expression and secretion of a polypeptide from a host cell ( e.g ., a leader sequence which functions as a secretory sequence for controlling transport of a polypeptide). The polypeptide can have the leader sequence cleaved by the host cell to form a “mature” form of the polypeptide.
[00217] In some embodiments, a polynucleotide comprises the coding sequence for a polypeptide fused in the same reading frame to a marker or tag sequence. For example, in some embodiments, a marker sequence is a hexa-histidine tag (SEQ ID NO:47) (HIS- tag) that allows for efficient purification of the polypeptide fused to the marker. In some embodiments, a marker sequence is a hemagglutinin (HA) tag derived from the influenza hemagglutinin protein when a mammalian host is used. In some embodiments, the marker sequence is a FLAG™ tag. In some embodiments, a marker may be used in conjunction with other markers or tags.
[00218] The present disclosure also provides variants of the polynucleotides described herein, wherein the variant encodes, for example, fragments, analogs, and/or derivatives of a polypeptide. In some embodiments, the present disclosure provides a polynucleotide comprising a polynucleotide having a nucleotide sequence at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, and in some embodiments, at least 96%, 97%, 98% or 99% identical to a polynucleotide sequence encoding a polypeptide described herein.
[00219] As used herein, the phrase “a polynucleotide having a nucleotide sequence at least 95% identical to a polynucleotide sequence” is intended to mean that the nucleotide sequence of the polynucleotide is identical to a reference sequence except that the polynucleotide sequence can include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence can be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence can be inserted into the reference sequence. It is understood by those of skill in the art that an appropriate calculation would be made for other “% identical” statements, for example, 90% identical or 85% identical. These mutations of the reference sequence can occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.
[00220] The polynucleotide variants can contain alterations in the coding regions, noncoding regions, or both. In some embodiments, a polynucleotide variant contains alterations which produce silent substitutions, additions, or deletions, but does not alter the properties or activities of the encoded polypeptide. In some embodiments, a polynucleotide variant comprises silent substitutions that results in no change to the amino acid sequence of the polypeptide (due to the degeneracy of the genetic code). In some embodiments, a polynucleotide variant comprises one or more mutated codons comprising one or more ( e.g ., 1, 2, or 3) substitutions to the codon that change the amino acid encoded by that codon. Methods for introducing one or more substitutions into a codon are known in the art, such as, e.g., PCR mutagenesis and site-directed mutagenesis. Polynucleotide variants can be produced for a variety of reasons, for example, to optimize codon expression for a particular host (e.g, change codons in the human mRNA to those preferred by a bacterial host such as E. coli). In some embodiments, a polynucleotide variant comprises at least one silent mutation in a noncoding or a coding region of the sequence.
[00221] In some embodiments, a polynucleotide variant is produced to modulate or alter expression (or expression levels) of the encoded polypeptide. In some embodiments, a polynucleotide variant is produced to increase expression of the encoded polypeptide. In some embodiments, a polynucleotide variant is produced to decrease expression of the encoded polypeptide. In some embodiments, a polynucleotide variant has increased expression of the encoded polypeptide as compared to a parental polynucleotide sequence. In some embodiments, a polynucleotide variant has decreased expression of the encoded polypeptide as compared to a parental polynucleotide sequence.
[00222] In some embodiments, a polynucleotide is isolated. In some embodiments, a polynucleotide is substantially pure.
[00223] Vectors and cells comprising each and every one of the polynucleotides described herein are also provided. In some embodiments, an expression vector comprises a polynucleotide or polynucleotides encoding a C3-binding agent described herein. In some embodiments, an expression vector or vectors comprise a polynucleotide or polynucleotides encoding a C3 -binding agent described herein. In some embodiments, an expression vector or vectors comprise a polynucleotide or polynucleotides encoding a polypeptide or polypepides that are part of a C3 -binding agent described herein. In some embodiments, a host cell comprises an expression vector or vectors comprising a polynucleotide or polynucleotides encoding a C3- binding agent described herein. In some embodiments, a host cell comprises an expression vector or vectors comprising a polynucleotide or polynucleotides encoding a polypeptide or polypeptides that are part of a C3 -binding agent described herein. In some embodiments, a host cell comprises a polynucleotide or polynucleotides encoding a C3 -binding agent described herein.
IV. Methods of use and pharmaceutical compositions
[00224] The present disclosure provides methods useful in a variety of applications including, but not limited to, therapeutic treatment methods, such as treatment of diseases or disorders associated with complement activation. The methods comprise use of a C3 -binding agent described herein. In some embodiments, a C3 -binding agent described herein is useful in methods for inhibiting complement activation. In some embodiments, a C3-binding agent described herein is useful in methods for inhibiting C3 activity. In some embodiments, a C3-binding agent described herein is useful in methods for treating respiratory illness. In some embodiments, a C3 -binding agent described herein is useful in methods for treating severe respiratory illness. In some embodiments, a C3-binding agent described herein is useful in methods of preventing or inhibiting the development of respiratory illness. In some embodiments, a C3 -binding agent described herein is useful in methods of preventing or inhibiting the development of severe respiratory illness. In some embodiments, a C3 -binding agent described herein is useful in methods of inhibiting complement pathway activation. In some embodiments, a C3-binding agent described herein is useful in methods of treating thrombosis. In some embodiments, a C3-binding agent described herein is useful in methods of preventing or inhibiting the development of thrombosis. In some embodiments, a C3-binding agent described herein is useful in methods of treating a coronavirus infection. In some embodiments, a C3-binding agent described herein is useful in methods of treating the sequelae of a coronavirus infection.
[00225] In some embodiments, a method of inhibiting complement activation in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of inhibiting complement activation of a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of inhibiting C3 activity in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein.
In some embodiments, a method of inhibiting C3 activity in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein.
[00226] In some embodiments, a method of treating a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3- binding agent described herein. In some embodiments, a method of treating a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of treating a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of treating a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting development of a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting development of a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting development of a respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting development of a severe respiratory illness in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments of the methods described herein, the severe respiratory illness is selected from the group consisting of: pneumonia, acute respiratory disease (ARD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), atypical ARDS, respiratory distress syndrome (RDS), severe acute respiratory syndrome (SARS),
Middle East respiratory syndrome (MERS), or coronavirus 2019 (COVID-19). In some embodiments of the methods described herein, the respiratory illness is pneumonia. In some embodiments of the methods described herein, the respiratory illness is ARD. In some embodiments of the methods described herein, the respiratory illness is ALI. In some embodiments of the methods described herein, the respiratory illness is ARDS.
The general definition of ARDS (Berlin definition 2012) is: ARDS is an acute diffuse, inflammatory lung injury, leading to increased pulmonary vascular permeability, increased lung weight, and loss of aerated lung tissue with hypoxemia and bilateral radiographic opacities, associated with increased venous admixture, increased physiological dead space and decreased lung compliance. The definition identifies three mutually exclusive categories of increasingly severe ARDS based on the degree of arterial hypoxemia as measured by the PaO2/FiO2 ratio (P/F): (i) mild - P/F 201 to 300 mm Hg; (ii) moderate - P/F 101 to 200 mm Hg; and (iii) severe - P/F < 100 mm Hg. In some embodiments of the methods described herein, the respiratory illness is atypical ARDS. Symptoms include hypoxia, shortness of breath, rapid breathing, and bluish skin coloration. For those who survive, a decreased quality of life is common. Atypical ARDS has been observed in some subjects infected with SARS-CoV-2. As used herein, “atypical ARDS” is characterized by moderate or severe hypoxia, without the feeling of shortness of breath. It is also characterized by severe hypoxia but well preserved lung gas volume, which is not seen in ARDS. In some embodiments of the methods described herein, the respiratory illness is RDS. In some embodiments of the methods described herein, the respiratory illness is SARS. In some embodiments of the methods described herein, the respiratory illness is MERS. In some embodiments of the methods described herein, the respiratory illness is COVID-19.
[00227] In some embodiments, a method of treating ARDS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating ARDS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of ARDS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of ARDS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed viral infection. In some embodiments, the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor.
In some embodiments, the viral infection is caused by a coronavirus, including but not limited to, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS- CoV-1, or Middle East respiratory syndrome coronavirus (MERS-CoV), optionally wherein the virus binds human ACE2 receptor. In some embodiments, the coronavirus is SARS-CoV-2.
[00228] In some embodiments, a method of inhibiting complement pathway activation in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of inhibiting complement pathway activation in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed viral infection. In some embodiments, the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor. In some embodiments, the viral infection is caused by a coronavirus, including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor. In some embodiments, the coronavirus is SARS-CoV-2.
[00229] In some embodiments, a method of treating thrombosis in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating thrombosis in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of thrombosis in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of thrombosis in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the thrombosis is a pulmonary embolism (PE), large blood vessel blood clot, a deep vein thrombosis (DVT), or microvascular thrombosis. In some embodiments, the thrombosis has the potential to lead to a stroke. In some embodiments, the subject has symptoms of a stroke. In some embodiments, the subject has been diagnosed with a stroke. In some embodiments, the subject has a suspected or confirmed viral infection. In some embodiments, the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor. In some embodiments, the viral infection is caused by a coronavirus, including but not limited to, SARS-CoV-2, SARS-CoV-1, or MERS-CoV, optionally wherein the virus binds human ACE2 receptor. In some embodiments, the coronavirus is SARS-CoV-2.
[00230] In some embodiments, a method of treating a coronavirus infection in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of treating a coronavirus infection in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the coronavirus, includes but is not limited to, SARS-CoV-2, SARS- CoV-1, or MERS-CoV. In some embodiments, the coronavirus is SARS-CoV-2. In some embodiments, the infection with SARS-CoV-2 results in the patient being diagnosed with COVID-19. In some embodiments, the infection with SARS-CoV-1 results in the patient being diagnosed with SARS. In some embodiments, the infection with MERS-CoV results in the patient being diagnosed with MERS.
[00231] In some embodiments, a method of treating SARS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating SARS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of SARS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of SARS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed SARS-CoV-1 infection.
[00232] In some embodiments, a method of treating MERS in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating MERS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of MERS in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of MERS in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed MERS-CoV infection.
[00233] In some embodiments, a method of treating COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of a C3-binding agent described herein. In some embodiments, a method of treating COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, a method of preventing or inhibiting the development of COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of a C3 -binding agent described herein. In some embodiments, a method of preventing or inhibiting the development of COVID-19 in a subject comprises administering to the subject a therapeutically effective amount of an anti-C3 antibody described herein. In some embodiments, the subject has a suspected or confirmed SARS-CoV-2 infection.
[00234] In some embodiments of the methods described herein, the viral infection is associated with a dysregulated pro-inflammatory cytokine response, which may be referred to as “severe cytokine release syndrome” or “cytokine storm”. In some embodiments, the pro-inflammatory cytokine response includes, but is not limited to, TNF-a, IL-6, IL-10, IL-la, IL-Ib, and IL-12. In some embodiments, there is an elevated level of cytokines in a sample obtained from a subject, wherein the cytokines include, but are not limited to, TNF-a, IL-6, IL-10, IL-la, IL-Ib, and IL-12. In some embodiments, there is an elevated level of C-reactive protein in a sample obtained from a subject.
[00235] In some embodiments of the methods described herein, the subject has lung damage, respiratory failure, kidney damage, kidney failure, liver damage, heart damage, vascular damage, thrombosis, stroke, central nervous system injury, and/or multiple organ failure. In some embodiments of the methods described herein, the subject has one or more symptoms selected from the group consisting of: hypoxemia, cough, wheezing, dyspnea, hyperpnea, pulmonary /lung inflammation, shortness of breath, labored breathing, rapid breathing, accumulation of alveolar fluid, pulmonary edema, vascular leakage, lymphocyte infiltration in the lung, lymphopenia, fever, chills, shaking chills, increased heart rate, chest pain, low blood pressure, headache, confusion, seizures, extreme tiredness, sepsis, bluish coloring of nails or lips, toe rashes/redness, toe swelling, loss of sense of smell, loss of sense of taste, and diarrhea.
[00236] In some embodiments of any of the methods described herein, the subject has mild, moderate or severe hypoxemia as determined by Partial Pressure of arterial oxygen/Fraction of inspired oxygen (PaCk/FiCk) or positive end-expiratory pressure (PEEP). In some embodiments of any of the methods described herein, the subject has severe hypoxemia with a Pa02/Fi02 of less than 100.
[00237] In some embodiments of the methods described herein, the subject is human. [00238] In some embodiments of the methods described herein, the C3 -binding agent has at least one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8,) of the following properties: (a) is an antagonist of C3; (b) inhibits C3 activity; (c) inhibits C3 cleavage; (d) inhibits C3 cleavage and C3a release; (e) inhibits complement activation; (f) inhibits activation of the alternative complement pathway; (g) inhibits activation of the classical complement pathway; and (h) inhibits activation of the alternative complement pathway and classical complement pathway.
[00239] In some embodiments of the methods described herein, the C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 38G10, e.g., as exemplified in Table 1 A. In some embodiments of the methods described herein, the C3 -binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10, e.g., as exemplified in Table IB. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(G56A), e.g., as exemplified in Table IB. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(G56T), e.g., as exemplified in Table 1C. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(N55E), e.g., as exemplified in Table 1C. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody Hz38G10(N55Q), e.g., as exemplified in Table 1C. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 3D8, e.g., as exemplified in Table 2. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 3G8, e.g., as exemplified in Table 3. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 1502, e.g., as exemplified in Table 4. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 27A8, e.g., as exemplified in Table 5. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 28C3, e.g., as exemplified in Table 6. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 38F5, e.g., as exemplified in Table 7. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 62B11, e.g., as exemplified in Table 8. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 62F2, e.g., as exemplified in Table 9. In some embodiments of the methods described herein, the C3-binding agent comprises a heavy chain variable region CDR1, CDR2, and CDR3 and a light chain variable region CDR1, CDR2, and CDR3 of antibody 63A3, e.g., as exemplified in Table 10.
[00240] In some embodiments of the methods described herein, the C3 -binding agent is an anti-C3 antibody. In some embodiments of the methods described herein, the anti- C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25) YIYPHNAGTTYNQQFTG (SEQ ID NO:27), YIYPHNTGTTYNQQFTG (SEQ ID NO:48), YIYPHEGGTTYNQQFTG (SEQ ID NO:52), or YIYPHQGGTTYNQQFTG (SEQ ID NO:56), and a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13); and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNGGTTYNQQFTG (SEQ ID NO:25), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNTGTTYNQQFTG (SEQ ID NO:48), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHEGGTTYNQQFTG (SEQ ID NO: 52), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14). In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHQGGTTYNQQFTG (SEQ ID NO:56), a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and (b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
[00241] In some embodiments of the methods described herein, the anti-C3 antibody comprises: (a) a heavy chain variable region of SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:222, SEQ ID NO:223, or SEQ ID NO:224; and (b) a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:33 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:34 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:222 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:223 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain variable region of SEQ ID NO:224 and a light chain variable region of SEQ ID NO:35. In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10. In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(G56A). In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(G56T). In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(N55E). In some embodiments of the methods described herein, the anti-C3 antibody is Hz38G10(N55Q).
[00242] In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:37. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:39. In some embodiments of the methods described herein, the anti-C3 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:37 and a light chain comprising the amino acid sequence of SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:39 and a light chain comprising the amino acid sequence of SEQ ID NO:41.
[00243] In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:37. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:39. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO:37 and a polypeptide of amino acids set forth in SEQ ID NO:41. In some embodiments of the methods described herein, the anti-C3 antibody comprises a polypeptide of amino acids set forth in SEQ ID NO: 39 and a polypeptide of amino acids set forth in SEQ ID NO:41.
[00244] In some embodiments of the methods described herein, a method comprises using a C3-binding agent in combination therapy. As used herein, combination therapy includes, but is not limited to, (i) combination with a medical device, for example, a ventilator or ECMO machine and (ii) combination with at least one additional therapeutic agent. In some embodiments, combination therapy comprises administration of a C3-binding agent in combination with at least one additional therapeutic agent and the usage of a medical device. In some embodiments of the methods described herein, a method comprises administering a C3-binding agent described herein in combination with usage of a medical device. In some embodiments of the methods described herein, a method comprises administering a C3-binding agent described herein in combination with at least one additional therapeutic agent.
Treatment with two or more therapeutic agents often uses agents that work by different mechanisms of action, although this is not required. Combination therapy using agents with different mechanisms of action may result in additive or synergetic effects. Combination therapy may allow for a lower dose of each agent than is used in monotherapy, thereby reducing toxic side effects and/or increasing the therapeutic index of the agent(s). Combination therapy may decrease the likelihood that resistance to an agent will develop.
[00245] In some embodiments, the combination of a C3 -binding agent described herein and at least one additional therapeutic agent results in additive or synergistic results. In some embodiments, the combination therapy results in an increase in the therapeutic index of the C3 -binding agent. In some embodiments, the combination therapy results in an increase in the therapeutic index of the additional therapeutic agent(s). In some embodiments, the combination therapy results in a decrease in the toxicity and/or side effects of the C3 -binding agent. In some embodiments, the combination therapy results in a decrease in the toxicity and/or side effects of the additional therapeutic agent(s). [00246] In some embodiments, a combination treatment comprises one additional therapeutic agent or two or more additional therapeutic agents. In some embodiments, treatment with a C3-binding agent can occur prior to, concurrently with, or subsequent to administration of the additional therapeutic agents. In some embodiments, combined administration includes co-administration, either in a single pharmaceutical formulation or using separate formulations, or consecutive administration in either order but generally within a time period such that all active agents can exert their biological activities. In some embodiments, preparation of agents and/or dosing schedules for additional therapeutic agents are according to manufacturers' instructions or as determined empirically by the skilled practitioner.
[00247] In some embodiments of the methods described herein, a C3-binding agent is administered to a subject in combination with one or more additional therapeutic agents. In some embodiments, an additional therapeutic agent is compstatin or an analog or derivative of compstatin ( e.g ., POT-4; APL-2, AMY- 101). In some embodiments, an additional therapeutic agent is a C5 inhibitor. In some embodiments, a C5 inhibitor is selected from the group including, but not limited to, eculizumab (SOLIRIS), LFG316, or Zimura (anti-C5 aptamer). In some embodiments, an additional therapeutic agent is a properdin inhibitor (e.g., an anti-properdin antibody). In some embodiments, an additional therapeutic agent is a Factor D inhibitor. In some embodiments, a Factor D inhibitor is an anti-Factor D antibody (e.g, lampalizumab). In some embodiments, an additional therapeutic agent is an anti-viral agent. In some embodiments, the anti-viral agent is remdisivir. In some instances, the additional therapeutic agent is one or more of: dexamethasone, remdesivir, baricitinib in combination with remdesivir, favipiravir, merimepodib, an anticoagulation drug selected from low-dose heparin or enoxaparin, bamlanivimab, a combination of bamlanivimab and etesevimab, a combination of casirivimab and imdevimab, convalescent plasma, an mRNA SARS-CoV-2 vaccine (e.g., those produced by Moderna and Pfizer), an attenuated SARS-CoV-2 virus vaccine, a dead SARS-CoV-2 virus vaccine, a viral vaccine against SARS-CoV-2 (e.g., an adenoviral vaccine such as those produced by Johnson & Johnson and Astrazeneca), an antibody or fragment thereof or small molecule that blocks interaction between hACE2 and the spike protein of SARS-CoV-2, protease inhibitors targeting SARS- CoV-2 S cleavage sites, viral fusion inhibitors, EK1C4, nelfmavir mesylate, furin inhibitors, or any other agent described in Huang et ak, Acta Pharmacol ogica Sinica (2020) 41:1141-1149; https://doi.org/10.1038/s41401-020-0485-4), or unmethylated CpG dinucleotides in combination with any of these agents.
[00248] It will be appreciated that the combination of a C3 -binding agent described herein and at least one additional therapeutic agent may be administered in any order or concurrently. In some embodiments, a C3-binding agent is administered to subjects that have previously undergone treatment with a therapeutic agent. In some embodiments, a C3-binding agent and a second therapeutic agent are administered substantially simultaneously or concurrently.
[00249] For the treatment of a disease, the appropriate dosage of a C3-binding agent of the present disclosure depends on the disorder or disease to be treated, the severity and course of the disorder or disease, the responsiveness of the disorder or disease, whether the agent is administered for therapeutic or preventative purposes, previous therapy, the subject’s condition, the subject's clinical history, and so on. A C3-binding agent can be administered one time or over a series of treatments lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. In some embodiments, the dose of a C3 -binding agent depends on the site of administration. In some embodiments, the dose of a C3-binding agent depends on the delivery system.
[00250] The present disclosure provides compositions comprising a C3-binding agent described herein. The present disclosure also provides pharmaceutical compositions comprising a C3-binding agent described herein and a pharmaceutically acceptable vehicle. In some embodiments, a pharmaceutical composition comprises an anti-C3 antibody described herein and a pharmaceutically acceptable vehicle. In some embodiments, a pharmaceutical composition comprises antibody Hz38G10 and a pharmaceutically acceptable vehicle. In some embodiments, a pharmaceutical composition comprises antibody Hz38G10(G56A) and a pharmaceutically acceptable vehicle.
[00251] Formulations are prepared for storage and use by combining a purified antibody or agent of the present disclosure with a pharmaceutically acceptable vehicle (i e.g ., a carrier or excipient). Those of skill in the art generally consider pharmaceutically acceptable carriers, excipients, and/or stabilizers to be inactive ingredients of a formulation or pharmaceutical composition.
[00252] Suitable pharmaceutically acceptable vehicles include, but are not limited to, nontoxic buffers such as phosphate, citrate, and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives such as octadecyldimethylbenzyl ammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens, such as methyl or propyl paraben, catechol, resorcinol, cyclohexanol, 3- pentanol, and m-cresol; low molecular weight polypeptides ( e.g ., less than about 10 amino acid residues); proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; carbohydrates such as monosaccharides, disaccharides, glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes such as Zn-protein complexes; and non-ionic surfactants such as TWEEN or polyethylene glycol. (. Remington : The Science and Practice of Pharmacy, 22nd Edition, 2012, Pharmaceutical Press, London.). In some embodiments, the formulation is in the form of an aqueous solution. In some embodiments, the formulation is stored in a lyophilized or in an alternative dried form. [00253] The binding agents of the present disclosure can be formulated in any suitable form for delivery to a target cell/tissue. In some embodiments, a C3-binding agent can be formulated as a liposome, microparticle, microcapsule, albumin microsphere, microemulsion, nano-particle, nanocapsule, or macroemulsion. In some embodiments, the pharmaceutical formulation includes an agent of the present disclosure complexed with liposomes. Methods to produce liposomes are known to those of skill in the art. For example, some liposomes can be generated by reverse phase evaporation with a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE).
[00254] In some embodiments, a C3-binding agent is formulated as a sustained-release preparation. Suitable examples of sustained-release preparations include semi- permeable matrices of solid hydrophobic polymers containing an agent, where the matrices are in the form of shaped articles (e.g., films or microcapsules). Sustained- release matrices include but are not limited to polyesters, hydrogels such as poly(2- hydroxy ethyl -methacrylate) or poly(vinyl alcohol), polylactides, copolymers ofL- glutamic acid and 7 ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid. [00255] The pharmaceutical compositions or formulations of the present disclosure can be administered in any number of ways for either local or systemic treatment. Administration can be (i) topical by epidermal or transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders, (ii) pulmonary by inhalation or insufflation of powders or aerosols, including by nebulizer, intratracheal, and intranasal, (iii) oral, or (iv) parenteral including intravenous, intraarterial, intratumoral, subcutaneous, intraperitoneal, intramuscular (e.g, injection or infusion), or intracranial (e.g, intrathecal or intraventricular).
EXAMPLES
Example 1
Generation of antibodies
[00256] Anti-C3 antibodies were generated using human C3 as the immunogen.
Single cell suspensions of lymphocytes were obtained from the spleens and lymph nodes of immunized mice after the individual animals had been determined to have suitable antibody titers. Lymphocytes were fused with murine myeloma cells by standard methods. Cells were dispersed into 96-well plates in HAT-containing selection media. Cell supernatants (approximately 10,000) were screened by ELISA for binding to human C3 and cynomolgus monkey (“cyno”) C3. Approximately 1600 cell clones were selected based on binding assay results.
Example 2
C3 Alternative Pathway Biochemical Assay
[00257] Anti-C3 antibodies identified in C3 binding assays were tested for their ability to inhibit the in vitro generation of C3a. C3a is produced by the cleavage of C3 by the C3 convertase complex. The C3a assay was based upon a previously described method (Loyet et al., 2014, J Pharmacol and Exp Ther\ 351:527-537). Briefly, 20 pL of solution of 50 nM human C3 (Complement Technologies) in GVB (bovine skin gelatin in veronal buffer) was mixed with 20 pL test anti-C3 antibodies or controls diluted in GVB for 10 minutes. Controls included (i) no C3; (ii) no Factor B and Factor D; (iii) no inhibitor; and (iv) compstatin as positive control. 100 nM human Factor B and human 100 nM Factor D (Complement Technologies) in 0.05 M EGTA/0.1 M MgC12/GVB were added to the C3/Ab samples and the mixtures were incubated for an additional 10 minutes. 20 pL of 50% rabbit serum (diluted 1 :2 in GVB) was added and incubated for 10 minutes, followed by addition of 20 pL of 0.5 M EDTA to quench the reaction. The addition of rabbit serum cleaves C3a produced by C3 convertase in the reaction mixtures to C3a des Arg (C3a des Arg has been found to be more stable than C3a). C3 des Arg concentrations were subsequently measured by ELISA. Briefly, a mouse anti-C3a/C3a des Arg antibody (Therm oFisher) was coated on ELISA plates at 1.0 pg/mL and incubated overnight at 4°C. The following day, the plates were washed and 60 pL of SuperBlock blocking buffer (Pierce) was added to the plates and incubated for 60 minutes at room temperature. Prior to adding samples, blocking buffer was decanted from the wells. Samples from the C3/Ab reactions were diluted, added to the plates, and incubated for 90 minutes. The plates were washed, biotinylated anti-human C3/C3a/C3a des Arg antibody (Biolegend) was added to the wells (0.5 pg/mL), and the plates were incubated for 60 minutes. The plates were washed, 20 pL poly-HRP streptavidin (Pierce) was added to each well and the plates were incubated for 30 minutes. For detection, SuperSignal PICO ELISA chemiluminescent substrate (ThermoFisher Scientific) was added to each well. The plates were read in a luminometer with a 0.2 second read time per well at 425 nm.
[00258] Under these assay conditions, test anti-C3 antibodies were ranked for their ability to inhibit C3a des Arg production. Several dozen exemplary antibodies were observed to strongly inhibit C3 cleavage and C3a release. The activities of three exemplary antibodies, 38G10, Ab2 (3D8), and Ab3 (15C12), at concentrations ranging from 0.1 to 1000 nM, are shown in Figure 1. Example 3
Alternative and classical pathway-dependent hemolysis assays
[00259] Hemolytic assays have traditionally been used to assess the functional activity of the complement system. For the alternative pathway (AP) assay, rabbit erythrocytes were included with C3-depleted human serum and 50 nM of human C3 in the presence of increasing concentrations of anti-C3 antibodies 38G10, Ab2, and Ab3. For the classical pathway (CP), IgM-coated sheep erythrocytes were incubated with C3- depleted human serum and increasing concentration of anti-C3 antibodies 38G10, Ab2, and Ab3 in the presence of 10 nM human C3. Hemolysis was determined by reading the absorbance of the supernatant at 412 nm. Results were expressed as a percentage of hemolysis observed in control samples without any inhibitor and can be calculated into IC50s. These assays were duplicated using cyno serum as a source of C3.
[00260] Results of the AP and CP assays using human C3 and cyno C3 are shown in Figure 2 for antibodies 38G10, Ab2 (3D8), and Ab3 (1502) (% hemolysis). The results are also summarized in Table 11.
[00261] These results showed that the anti-C3 antibodies inhibited activation of both the alternative and classical pathways as demonstrated by the inhibition of hemolysis.
Table 11
Figure imgf000157_0001
Example 4
Binding Affinity
[00263] The binding affinities of exemplary anti-C3 antibodies to human C3 and cyno C3 were measured using a Biacore system (GE Healthcare LifeSciences). Briefly, anti- Fc antibody (Sigma-Aldrich) was immobilized on all four flow cells of a CM5 chip using amine coupling reagents (GE Healthcare LifeSciences). Antibodies were captured on flow cells 2, 3, and 4 using flow cell 1 as a reference. Concentrations ranging from 0.62 - 40 nM of human C3 or cyno C3 were injected at a flow rate of 50 μL/min at 37°C. Kinetic data were collected over time and fit using the simultaneous global fit equation to yield affinity constants (KD values) for each antibody.
[00264] Antibody 38G10 bound to human C3 with a KD of 1.9 x 10'10 M and to cyno C3 with a KD of 1.6 X lO-10 M. [00265] Table 12 shows the binding affinities for antibodies 38G10, 3D8, and 15C12.
Table 12
Figure imgf000158_0001
Example 5
Binding Profile
[00266] The anti-C3 antibodies described herein were characterized for their ability to bind a number of other components of the complement cascade. Antibodies were screened for binding to C3, C3a, C3b, C3c, C3d, iC3b, and Factor Bb. A representative set of the results is shown in Table 13. All the anti-C3 antibodies included in Table 13 were able to block complement activation. The data are relative to the binding of the negative control mIgG2 antibody.
Table 13
Figure imgf000159_0001
[00267] These data indicated that antibody 38G10 had a unique C3 binding profile, in that it bound preferentially to C3 with no detectable or very limited binding to any of the fragments of the complement cascade.
Example 6
Characterization and humanization of anti-C3 antibodies
[00268] Antibody 38G10 was humanized by methods known to those skilled in the art and humanized 38G10 is referred to herein as Hz38G10. Antibody 38G10 was found to have a potential glycosylation site in CDR2 of the heavy chain variable region, YIYPHNGGTTYNONFTG (SEQ ID NO: 10). The consensus glycosylation site for N- linked glycans is N-X-S/T, wherein X can be any amino acid except proline. During the humanization process, the heavy chain CDR2 was reengineered to remove the glycosylation site resulting in antibody Hz38G10 comprising a heavy chain CDR2 comprising YIYPHNGGTTYNQQFTG (SEQ ID NO:25). The heavy chain variable sequence of antibody 38G10 is SEQ ID NO:31 and the light chain variable sequence is SEQ ID NO:32; the heavy chain variable sequence of antibody Hz38G10 is SEQ ID NO:33 and the light chain variable sequence is SEQ ID NO:35; CDRs for 38G10 and Hz38G10 are disclosed in Tables 1A and IB.
Example 7
Characterization of Hz38G10
[00269] The binding affinity of Hz38G10 was determined using a Biacore system as described herein and compared with the binding affinity of the parental 38G10 antibody. Antibody Hz38G10 had a binding affinity to human C3 of 3 x 10'10 M and to cyno C3 of 1.8 x 10'9 M as compared to the parental antibody’s binding affinity of approximately 1 x 10'10 M to both human and cyno C3 (at 37°C). These results demonstrated that the humanization process for antibody 38G10, as well as the removal of a glycosylation site within CDR2 of the heavy chain, did not have a significant effect on the binding capabilities to human C3 or cyno C3.
[00270] Due to a lack of anti-C3 antibodies in the clinic or licensed by the FDA, the binding affinity of Hz38G10 to human C3 was compared against compstatin, a 13- residue cyclic peptide with potent C3 inhibitory activity that was identified several years ago (Morikis et ah, 1998, Protein Science ; 7:619-627). More recently, a number of compstatin analogs and derivatives have been generated and studied. The binding affinity of Hz38G10 to human C3 was compared to the binding affinity of a compstatin derivative (referred to herein as “COMP”) and a pegylated version of the compstatin derivative (referred to herein as “COMP -PEG”). A Biacore system was used to assess the binding affinities of the different molecules to human C3. Binding affinity of Hz38G10 to human C3 at 25°C and 37°C was carried out as described in Example 4. Evaluation of COMP or COMP -PEG binding to human C3 was carried out as follows. Human C3 was immobilized on flow cell 2 of a CM5 chip using amine-coupling reagents (GE Healthcare LifeSciences) and flow cell 1 (immobilized with a similar amount of human IgG) was used as a reference. Concentrations ranging from 1.56 - lOOnM of COMP were injected at a flow rate of 50 pL/min for evaluation of binding affinity at 25°C and 37°C. Similar experiments were set up with COMP-PEG (concentration range of 25 - 400 nM) for evaluation at 37°C. COMP -PEG binding to human C3 was also evaluated at 25°C with modifications. Briefly, rat anti-PEG antibody (Abeam Inc.) was immobilized on a CM5 chip surface using amine coupling reagents and COMP-PEG was captured on flow cell 2. Human C3 (concentration range 12 - 100 nM) was injected at a flow rate of 50 pL/min. Kinetic data were collected over time and fit using the simultaneous global fit equation to yield affinity constants (KD values) for each antibody.
[00271] A set of representative results is shown in Table 14.
Table 14
Figure imgf000161_0001
[00272] These results suggest that antibody Hz38G10 binds to human C3 with a significantly greater affinity than a compstatin derivative or the pegylated version (ranging from approximately 50 - 80 times greater).
[00273] To confirm that the humanized version of antibody 38G10 retained the ability to inhibit complement activation, Hz38G10 was assessed in alternative and classical pathway-dependent hemolysis assays as described herein and compared to COMP and COMP -PEG. Hz38G10 and COMP or antibody Hz38G10 and COMP -PEG were tested for their respective abilities to inhibit hemolysis in the assays.
[00274] As shown in Figures 3 and 4 and Table 15, the results of these experiments indicated that antibody Hz38G10 was more effective at inhibiting complement activation than a compstatin derivative or a pegylated version of the compstatin derivative. Antibody Hz38G10 had similar inhibitory activity on both the classical and alternative pathways. In contrast, COMP and COMP -PEG appeared to have greater activity on the alternative pathway than the classical pathway. Antibody Hz38G10 was approximately 10-fold more effective than COMP or COMP -PEG in inhibiting the alternative pathway and approximately 40- to 60-fold more effective than the compstatin derivatives on inhibiting the classical pathway. Table 15
Figure imgf000162_0001
[00275] An additional experiment was undertaken to determine whether antibody Hz38G10 and the compstatin derivative (COMP) bound to the same or similar region on human C3. COMP was immobilized on a CM5 chip using amine coupling reagents (GE Healthcare LifeSciences) at a high density (approx. 3000 RUs). In separate reactions in a 96-well plate, human C3 (5 nM) was titrated with increasing concentrations of antibody Hz38G10 or COMP (0.05 - 100 nM). After an incubation of approximately 3 hours at room temperature, the C3-COMP or C3-Hz38G10 reactions were injected over the COMP-coated surface.
[00276] As shown in Figure 5, increasing concentrations of COMP reduced binding of C3 to COMP surface due to self-blocking (positive control). However, increasing concentration of Hz38G10 did not prevent C3 from binding to the COMP-coated chip surface. The results of this experiment indicated that antibody Hz38G10 and COMP bind different epitopes on human C3.
Example 8
Generation and characterization of Hz38G10 variants
[00277] Sequence analysis of the heavy chain CDRs of antibody Hz38G10 identified a potential spot for deamidation within CDR2, YIYPHNGGTTYNQQFTG (SEQ ID NO:25). An elevated pH stress test was performed at pH 8.5 and deamidation at the asparagine (N55 of SEQ ID NO:33) was determined to be 29.75% on Day 0 and 48.23% on Day 7 (an increase of 18.5%). A variant of antibody Hz38G10 was generated with a N55D mutation in the heavy chain CDR2, a change that would mimic a fully deamidated molecule. This variant was assayed for binding to human C3 using a Biacore system as described herein and was shown to have reduced binding as compared to Hz38G10.
[00278] Four additional Hz38G10 variants were generated with amino acid substitutions within the heavy chain CDR2 at position N55 or G56 of SEQ ID NO:33. The Hz38G10 variants contained (i) a N55E mutation; (ii) a N55Q mutation; (iii) a G56A mutation; or (iv) a G56T mutation. The heavy chain variable region sequence of antibody Hz38G10(G56A) is SEQ ID NO:34, Hz38G10(G56T) is SEQ ID NO:222, Hz38G10(N55E) is SEQ ID NO:223, and Hz38G10(N55Q) is SEQ ID NO:224; the light chain variable region sequence for all variants is SEQ ID NO:35; CDRs are disclosed in Tables 1B-1C.
[00279] These antibodies were assayed for binding to human C3 using a Biacore system as described herein and compared to antibody Hz38G10. The results are shown in Table 16.
Table 16
Figure imgf000163_0001
[00280] These results indicated that a mutation at position G56 retained binding affinity to a greater extent than a mutation at position N55.
[00281] To determine if the substitution of alanine for glycine at position 56 of the heavy chain variable region (SEQ ID NO:33) reduced deamidation within the CDR2
(YIYPHNAGTTYNQQFTG - SEQ ID NO: 27), a pH stress test was performed with antibody Hz38G10(G56A) as described above. For this variant, deamidation at N55 of Hz38G10(G56A) (SEQ ID NO:34) was determined to be 0.66% on Day 0 and 1.0% on Day 7. These results showed that antibody Hz38G10(G56A) was very stable under deamidation stressed conditions.
[00282] To determine whether antibody Hz38G10(G56A) retained the ability to inhibit complement activation, the antibody was assessed in alternative and classical pathway hemolysis assays as described herein and compared to antibody Hz38G10. Hz38G10 and Hz38G10(G56A) were tested for their ability to inhibit hemolysis in the assays. [00283] As shown in Table 17, antibody Hz38G10(G56A) showed almost identical inhibition activity when compared to antibody Hz38G10 for human C3 and cyno C3.
Table 17
Figure imgf000164_0001
Example 9
Generation of Hz38G10 scFv
[00284] A scFv version of an exemplary anti-C3 antibody was generated. scFv molecules based on the heavy chain variable region (VH) and the light chain variable region (VL) of Hz38G10 were designed in two orientations and with or without disulfide stabilization. The four molecules were (i) VL-linker-VH scFv, (ii) VH-linker- VL scFv, (iii) VL-linker-VH dsscFv; and (iv) VH-linker-VL dsscFv, wherein dsscFv indicates a scFv with an engineered disulfide bond between the VL and VH regions. These four molecules were tested in an E.coli expression system and based on several parameters, for example, expression titer and percent monomer, the VL-linker-VH dsscFv molecule was selected for additional studies and characterization. The sequence of Hz38G10 dsscFv is disclosed herein as SEQ ID NO:228.
[00285] Hz38G10 dsscFv antibodies were produced via expression as inclusion bodies. BL21(DE3) E. coli were transformed with a plasmid encoding Hz38G10 VL-linker-VH dsscFv, referred to hereafter as Hz38G10 dsscFv. Transformed BL21(DE3) E. coli were harvested by low speed centrifugation. The bacterial pellet was resuspended by magnetic stirring in 50 mM Tris-HCl pH 7.5 for 15 minutes at room temperature. For cell disruption, the suspension was passed 3 times through a microfluidizer at 200 Bar. Inclusion bodies were recovered from broken cells by centrifugation at 4°C in a fixed angle rotor operating at 13,000 xg for 45 minutes. The resulting inclusion body pellet was washed with cold Milli-Q® water until the lipid and cell wall layers were removed. The washed inclusion body pellet was resuspended in 5 volumes of Milli-Q® water by magnetic stirring and collected by centrifugation at 13,000 xg for 30 minutes. Washed inclusion bodies were solubilized in 10 volumes of 8 M guanidine-HCl, 50 mM Tris- HCl pH 7.5, 1 mM TCEP using magnetic stirring and incubated overnight at 4°C. The following day, a dounce homogenizer with loose pestle was used to resuspend any remaining solid inclusion body particles. Solubilized inclusion bodies were diluted dropwise 1 :25 into refold buffer (3 M urea, 50 mM Tris-HCl pH 8.0, 160 mM L- arginine, 1 mM cysteine, 3 mM cystamine) under gentle magnetic stirring at room temperature to a final concentration of 0.1 mg/ml. Refolding was carried out for 48 hours at 4°C and disulfide bond formation was monitored by SDS-PAGE under nonreducing conditions. The solution containing the refolded dsscFv was concentrated 5- fold by ultrafiltration/diafiltration (UF/DF) on a 0.22 m2 10,000 DaNMWCO tangential flow filtration membrane. The concentrate was cleared of misfolded dsscFv by rapid acidification to pH 2.8 with 4 volumes of 100 mM glycine pH 2.5. The solution was held at pH 2.8 as the solution was further concentrated 5-fold. After 15 minutes, the pH was quickly returned to neutral with addition of concentrated Tris-HCl solution. The resulting acid-treated solution was buffer-exchanged with diavolumes of Protein-L affinity binding buffer (50 mM Tris-HCl pH 7.5, 50 mM NaCl). Recovered diafilitration product was cleared of precipitate by centrifugation at 13,000 xg for 45 minutes at 4°C and subsequently filtered using a 0.2 pm membrane.
[00286] Hz38G10 dsscFv antibodies were purified in two chromatography steps: (i) a Protein L affinity column for capture and (ii) a cation exchange (CEX) column for polishing. For Protein L affinity chromatography, the suspension containing Hz38G10 dsscFv was applied to a 5 mL MiniChrom TOYOPEARL® AF-rProtein L-650F column (Tosoh Bioscience), washed extensively with Tris-buffered saline, and eluted with a step gradient of 100 mM glycine pH 2.5. The resulting elution pool (Pool M) was dialyzed against CEX binding buffer (20 mM sodium acetate pH 5.0). Dialyzed Pool M was applied to a HiTrap® SP HP cation exchange column (GE Healthcare), washed with 20 mM sodium acetate, pH 5.0, and eluted with an isocratic gradient of 50 sodium chloride in 20 mM sodium acetate, pH 5.0. Hz38G10 dsscFv-enriched fractions were pooled and evaluated by SDS-PAGE and analytical SEC methods. Subsequently, a Hz38G10(G56A) dsscFv antibody (SEQ ID NO:229) was generated following the same general methods.
Example 10
Characterization of Hz38G10 dsscFv
[00287] The solubility parameters of Hz38G10 dsscFv were investigated. The purified antibodies were dialyzed into buffer consisting of 10 mM sodium phosphate, 40 mM sodium chloride, 0.03% polysorbate 20, and 5% sucrose, pH 6.2. The solution was micro-concentrated using 10,000 Da NMWCO spin concentrator at 3,000 xg at room temperature. Protein concentration was measured on an Agilent Cary 60 UV-VIS Solo VPE spectrophotometer. Protein concentration reached 229 mg/ml with no visible aggregation.
[00288] Hz38G10 dsscFv were compared to Hz38G10 intact antibody and a Fab version of Hz38G10 in several assays. Binding affinity to human C3 was assessed using a Biacore system as described herein. In addition, inhibition of the alternative pathway and classical pathway by the three antibodies was assessed in hemolysis assays as described herein. [00289] The results are summarized in Table 18.
Table 18
Figure imgf000167_0001
[00290] These results showed that Hz38G10 dsscFv had a similar KD as compared to the intact antibody. In addition, Hz38G10 dsscFv had a similar level of inhibition of hemolytic activity in both the AP and CP pathways as compared to the intact antibody or the Fab version. It should be noted that the intact Hz38G10 antibody has two antigen-binding sites, while the Fab and dsscFv antibodies have only one antigenbinding site.
[00291] Although the foregoing present disclosure has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the present disclosure. The embodiments of the present disclosure described herein are intended to be merely exemplary, and those skilled in the art will recognize numerous equivalents to the specific procedures described herein. All such equivalents are considered to be within the scope of the present disclosure and are covered by the embodiments.
[00292] All publications,, patents, patent applications, internet sites, and accession numbers/database sequences including both polynucleotide and polypeptide sequences cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, internet site, or accession number/database sequence were specifically and individually indicated to be so incorporated by reference. [00293] Following are sequences disclosed in the application. CDR sequences are listed in Tables 1A and IB.
Human C3 amino acid sequence with predicted signal sequence underlined (SEQ ID NO:l)
MGPTSGPSLLLLLLTHLPLALGSPMYS 11TPNILRLESEETMVLEAHDAQGDVPVTVT VHDFPGKKLVLSSEKTVLTPATNHMGNVTFTIPANRE FKSEKGRNKFVTVQATFGTQV VEKVVLVSLQSGYLFIQTDKTIYTPGSTVLYRI FTVNHKLLPVGRTVMVNIENPEGIP VKQDSLSSQNQLGVLPLSWDIPELVNMGQWKIRAYYENSPQQVFSTEFEVKEYVLPSF EVIVEPTEKFYYIYNEKGLEVTITARFLYGKKVEGTAFVI FGIQDGEQRISLPESLKR IPIEDGSGEW LSRKVLLDGVQNPRAEDLVGKSLYVSATVILHSGSDMVQAERSGIPI VTSPYQIHFTKTPKYFKPGMPFDLMVFVTNPDGSPAYRVPVAVQGEDTVQSLTQGDGV AKLSINTHPSQKPLSITVRTKKQELSEAEQATRTMQALPYSTVGNSNNYLHLSVLRTE LRPGETLNVNFLLRMDRAHEAKIRYYTYLIMNKGRLLKAGRQVREPGQDLW LPLSIT TDFIPSFRLVAYYTLIGASGQREW ADSVWVDVKDSCVGSLW KSGQSEDRQPVPGQQ MTLKIEGDHGARW LVAVDKGVFVLNKKNKLTQSKIWDW EKADIGCTPGSGKDYAGV FSDAGLTFTSSSGQQTAQRAELQCPQPAARRRRSVQLTEKRMDKVGKYPKELRKCCED GMRENPMRFSCQRRTRFISLGEACKKVFLDCCNYITELRRQHARASHLGLARSNLDED IIAEENIVSRSEFPESWLWNVEDLKEPPKNGISTKLMNI FLKDSITTWEILAVSMSDK KGICVADPFEVTVMQDFFIDLRLPYSW RNEQVEIRAVLYNYRQNQELKVRVELLHNP AFCSLATTKRRHQQTVTIPPKSSLSVPYVIVPLKTGLQEVEVKAAVYHHFISDGVRKS LKW PEGIRMNKTVAVRTLDPERLGREGVQKEDIPPADLSDQVPDTESETRILLQGTP VAQMTEDAVDAERLKHLIVTPSGCGEQNMIGMTPTVIAVHYLDETEQWEKFGLEKRQG ALELIKKGYTQQLAFRQPSSAFAAFVKRAPSTWLTAYW KVFSLAVNLIAIDSQVLCG AVKWLILEKQKPDGVFQEDAPVIHQEMIGGLRNNNEKDMALTAFVL ISLQEAKDICEE QVNSLPGSITKAGDFLEANYMNLQRSYTVAIAGYALAQMGRLKGPLLNKFLTTAKDKN RWEDPGKQLYNVEATSYALLALLQLKDFDFVPPW RWLNEQRYYGGGYGSTQATEMVF QALAQYQKDAPDHQELNLDVSLQLPSRSSKITHRIHWESASLLRSEETKENEGFTVTA EGKGQGTLSW TMYHAKAKDQLTCNKFDLKVTIKPAPETEKRPQDAKNTMILEICTRY RGDQDATMSILDISMMTGFAPDTDDLKQLANGVDRYISKYELDKAFSDRNTLI IYLDK VSHSEDDCLAFKVHQYFNVELIQPGAVKVYAYYNLEESCTRFYHPEKEDGKLNKLCRD ELCRCAEENCFIQKSDDKVTLEERLDKACEPGVDYVYKTRLVKVQLSNDFDEYIMAIE QTIKSGSDEVQVGQQRTFISPIKCREALKLEEKKHYLMWGLSSDFWGEKPNLSYI IGK DTWVEHWPEEDECQDEENQKQCQDLGAFTESMW FGCPN
Human C3 amino acid sequence without predicted signal sequence (SEQ ID NO:2)
SPMYSIITPNILRLESEETMVLEAHDAQGDVPVTVTVHDFPGKKLVLSSEKTVLTPAT NHMGNVTFTIPANREFKSEKGRNKFVTVQATFGTQW EKW LVSLQSGYLFIQTDKTI YTPGSTVLYRIFTVNHKLLPVGRTVMVNIENPEGIPVKQDSLSSQNQLGVLPLSWDIP ELVNMGQWKIRAYYENSPQQVFSTEFEVKEYVLPSFEVIVEPTEKFYYI YNEKGLEVT ITARFLYGKKVEGTAFVIFGIQDGEQRISLPESLKRIPIEDGSGEVVLSRKVLLDGVQ NPRAEDLVGKSLYVSATVILHSGSDMVQAERSGIPIVTSPYQIHFTKTPKYFKPGMPF DLMVFVTNPDGSPAYRVPVAVQGEDTVQSLTQGDGVAKLS INTHPSQKPLSITVRTKK QELSEAEQATRTMQALPYSTVGNSNNYLHLSVLRTELRPGETLNVNFLLRMDRAHEAK IRYYTYLIMNKGRLLKAGRQVREPGQDLW LPLSITTDFIPSFRLVAYYTLIGASGQR EW ADSVWVDVKDSCVGSLW KSGQSEDRQPVPGQQMTLKIEGDHGARW LVAVDKGV FVLNKKNKLTQSKIWDW EKADIGCTPGSGKDYAGVFSDAGLTFTSSSGQQTAQRAEL QCPQPAARRRRSVQLTEKRMDKVGKYPKELRKCCEDGMRENPMRFSCQRRTRFISLGE ACKKVFLDCCNYITELRRQHARASHLGLARSNLDEDI IAEENIVSRSEFPESWLWNVE DLKEPPKNGISTKLMNIFLKDSITTWEILAVSMSDKKGICVADPFEVTVMQDFFIDLR LPYSW RNEQVEIRAVLYNYRQNQELKVRVELLHNPAFCSLATTKRRHQQTVTIPPKS SLSVPYVIVPLKTGLQEVEVKAAVYHHFISDGVRKSLKW PEGIRMNKTVAVRTLDPE RLGREGVQKEDIPPADLSDQVPDTESETRILLQGTPVAQMTEDAVDAERLKHLIVTPS GCGEQNMIGMTPTVIAVHYLDETEQWEKFGLEKRQGALELIKKGYTQQLAFRQPSSAF AAFVKRAPSTWLTAYW KVFSLAVNLIAIDSQVLCGAVKWLILEKQKPDGVFQEDAPV IHQEMIGGLRNNNEKDMALTAFVLISLQEAKDICEEQVNSLPGS ITKAGDFLEANYMN LQRSYTVAIAGYALAQMGRLKGPLLNKFLTTAKDKNRWEDPGKQLYNVEATSYALLAL LQLKDFDFVPPW RWLNEQRYYGGGYGSTQATEMVFQALAQYQKDAPDHQELNLDVSL QLPSRSSKITHRIHWESASLLRSEETKENEGFTVTAEGKGQGTLSW TMYHAKAKDQL TCNKFDLKVTIKPAPETEKRPQDAKNTMILEICTRYRGDQDATMS ILDISMMTGFAPD TDDLKQLANGVDRYISKYELDKAFSDRNTLI IYLDKVSHSEDDCLAFKVHQYFNVELI QPGAVKVYAYYNLEESCTRFYHPEKEDGKLNKLCRDELCRCAEENCFIQKSDDKVTLE ERLDKACEPGVDYVYKTRLVKVQLSNDFDEYIMAIEQTIKSGSDEVQVGQQRTFISPI KCREALKLEEKKHYLMWGLSSDFWGEKPNLSYI IGKDTWVEHWPEEDECQDEENQKQC QDLGAFTESMW FGCPN
Human C3 and C3b beta chain (amino acids 23-667) (SEQ ID NO:3)
SPMYSIITPNILRLESEETMVLEAHDAQGDVPVTVTVHDFPGKKLVLSSEKTVLTPAT NHMGNVTFTIPANREFKSEKGRNKFVTVQATFGTQW EKW LVSLQSGYLFIQTDKTI YTPGSTVLYRIFTVNHKLLPVGRTVMVNIENPEGIPVKQDSLSSQNQLGVLPLSWDIP ELVNMGQWKIRAYYENSPQQVFSTEFEVKEYVLPSFEVIVEPTEKFYYI YNEKGLEVT ITARFLYGKKVEGTAFVIFGIQDGEQRISLPESLKRIPIEDGSGEVVLSRKVLLDGVQ NPRAEDLVGKSLYVSATVILHSGSDMVQAERSGIPIVTSPYQIHFTKTPKYFKPGMPF DLMVFVTNPDGSPAYRVPVAVQGEDTVQSLTQGDGVAKLS INTHPSQKPLSITVRTKK QELSEAEQATRTMQALPYSTVGNSNNYLHLSVLRTELRPGETLNVNFLLRMDRAHEAK IRYYTYLIMNKGRLLKAGRQVREPGQDLW LPLSITTDFIPSFRLVAYYTLIGASGQR EW ADSVWVDVKDSCVGSLW KSGQSEDRQPVPGQQMTLKIEGDHGARW LVAVDKGV FVLNKKNKLTQSKIWDW EKADIGCTPGSGKDYAGVFSDAGLTFTSSSGQQTAQRAEL QCPQPAA
Human C3 alpha chain (amino acids 672-1663) (SEQ ID NO:4)
SVQLTEKRMDKVGKYPKELRKCCEDGMRENPMRFSCQRRTRFISLGEACKKVFLDCCN
YITELRRQHARASHLGLARSNLDEDI IAEENIVSRSEFPESWLWNVEDLKEPPKNGIS
TKLMNIFLKDSITTWEILAVSMSDKKGICVADPFEVTVMQDFFIDLRLPYSW RNEQV
EIRAVLYNYRQNQELKVRVELLHNPAFCSLATTKRRHQQTVTIPPKSSLSVPYVIVPL
KTGLQEVEVKAAVYHHFISDGVRKSLKW PEGIRMNKTVAVRTLDPERLGREGVQKED
IPPADLSDQVPDTESETRILLQGTPVAQMTEDAVDAERLKHLIVTPSGCGEQNMIGMT
PTVIAVHYLDETEQWEKFGLEKRQGALELIKKGYTQQLAFRQPSSAFAAFVKRAPSTW LTAYW KVFSLAVNLIAIDSQVLCGAVKWLILEKQKPDGVFQEDAPVIHQEMIGGLRN NNEKDMALTAFVLISLQEAKDICEEQVNSLPGS ITKAGDFLEANYMNLQRSYTVAIAG YALAQMGRLKGPLLNKFLTTAKDKNRWEDPGKQLYNVEATSYALLALLQLKDFDFVPP W RWLNEQRYYGGGYGSTQATEMVFQALAQYQKDAPDHQELNLDVSLQLPSRSSKITH RIHWESASLLRSEETKENEGFTVTAEGKGQGTLSW TMYHAKAKDQLTCNKFDLKVTI KPAPETEKRPQDAKNTMILEICTRYRGDQDATMS ILDISMMTGFAPDTDDLKQLANGV DRYISKYELDKAFSDRNTLIIYLDKVSHSEDDCLAFKVHQYFNVELIQPGAVKVYAYY NLEESCTRFYHPEKEDGKLNKLCRDELCRCAEENCFIQKSDDKVTLEERLDKACEPGV DYVYKTRLVKVQLSNDFDEYIMAIEQTIKSGSDEVQVGQQRTFISPIKCREALKLEEK KHYLMWGLSSDFWGEKPNLSYIIGKDTWVEHWPEEDECQDEENQKQCQDLGAFTESMV VFGCPN
Human C3b alpha chain (amino acids 749-1663) (SEQ ID NO:5)
SNLDEDIIAEENIVSRSEFPESWLWNVEDLKEPPKNGISTKLMNI FLKDSITTWEILA VSMSDKKGICVADPFEVTVMQDFFIDLRLPYSW RNEQVEIRAVLYNYRQNQELKVRV ELLHNPAFCSLATTKRRHQQTVTIPPKSSLSVPYVIVPLKTGLQEVEVKAAVYHHFIS DGVRKSLKW PEGIRMNKTVAVRTLDPERLGREGVQKEDIPPADLSDQVPDTESETRI LLQGTPVAQMTEDAVDAERLKHLIVTPSGCGEQNMIGMTPTVIAVHYLDETEQWEKFG LEKRQGALELIKKGYTQQLAFRQPSSAFAAFVKRAPSTWLTAYW KVFSLAVNLIAID SQVLCGAVKWLILEKQKPDGVFQEDAPVIHQEMI GGLRNNNEKDMALTAFVLISLQEA KDICEEQVNSLPGSITKAGDFLEANYMNLQRSYTVAIAGYALAQMGRLKGPLLNKFLT TAKDKNRWEDPGKQLYNVEATSYALLALLQLKDFDFVPPW RWLNEQRYYGGGYGSTQ ATEMVFQALAQYQKDAPDHQELNLDVSLQLPSRSSKITHRIHWESASLLRSEETKENE GFTVTAEGKGQGTLSW TMYHAKAKDQLTCNKFDLKVTIKPAPETEKRPQDAKNTMIL EICTRYRGDQDATMSILDISMMTGFAPDTDDLKQLANGVDRYISKYELDKAFSDRNTL IIYLDKVSHSEDDCLAFKVHQYFNVELIQPGAVKVYAYYNLEESCTRFYHPEKEDGKL NKLCRDELCRCAEENCFIQKSDDKVTLEERLDKACEPGVDYVYKTRLVKVQLSNDFDE YIMAIEQTIKSGSDEVQVGQQRTFISPIKCREALKLEEKKHYLMWGLSSDFWGEKPNL SYIIGKDTWVEHWPEEDECQDEENQKQCQDLGAFTESMW FGCPN
Human C3a (amino acids 672-748) (SEQ ID NO:6)
SVQLTEKRMDKVGKYPKELRKCCEDGMRENPMRFSCQRRTRFISLGEACKKVFLDCCN
YITELRRQHARASHLGLAR
Cynomolgus monkey C3 amino acid sequence with predicted signal sequence underlined (SEQ ID NO:7)
MGLTSGPSLLLLLLIHLPLALGTPMYSM ITPNVLRLESEETVVLEAHDANGDVPVTVT VHDFPGKKLVLSSEKTVLTPATSHMGSVTIRIPANKE FKSEKGHNKFVTVQATFGAQV VEKW LVSLQSGYLFIQTDKTIYTPGSTVLCRIFTVNHKLLPVGRTVW NIENPDGIP VKQDSLSSQNQFGILPLSWDIPELVNMGQWKIRAYYENSPQQVFSTEFEVKEYVLPSF EVIVEPTEKFYYIYNQKGLEVTITARFLYGKKVEGTAFVI FGIQDGEQRISLPESLKR IQIEDGSGDAVLSRKVLLDGVQNPRPEDLVGKSLYVSVTVILHSGSDMVQAERSGIPI VTSPYQIHFTKTPKYFKPGMPFDLMVFVTNPDGSPAYRVPVAVQGEDAVQSLTQGDGV AKLSINTHPSQKPLSITVRTKKRELSEAEQATRTMEAQPYSTVGNSNNYLHLSVPRAE LRPGETLNVNFLLRMDRTQEAKIRYYTYLIMNKGKLLKVGRQVREPGQDLW LPLSIT TDFIPSFRLVAYYTLIGANGQREW ADSVWVDVKDSCVGSLW KSGQSEDRQPLPGQQ MTLKIEGDHGARVGLVAVDKGVFVLNKKNKLTQSKIWDW EKADIGCTPGSGKDYAGV FSDAGLTFASSSGQQTAQRAELQCPQPAARRRRSVQLAEKRMDKVGQYPKELRKCCEH GMRENPMRFSCQRRTRYITLDEACKKAFLDCCNYITELRRQHARASHLGLARSNLDED IIAEENIVSRSEFPESWLWKIEELKEAPKNGISTKLMNI FLKDSITTWEILAVSLSDK KGICVADPFEVTVMQDFFIDLRLPYSW RNEQVEIRAVLYNYRQNQELKVRVELLHNP AFCSLATAKRRHQQTVTIPPKSSLSVPYVIVPLKTGQQEVEVKAAVYHFFISDGVRKS LKW PEGIRMNKTVAVRTLDPERLGQEGVQREDVPPADLSDQVPDTESETRILLQGTP VAQMTEDAIDAERLKHLIVTPSGCGEQNMITMTPTVIAVHYLDETEQWEKFGPEKRQG ALELIKKGYTQQLAFRQPSSAFAAFLNRAPSTWLTAYW KVFSLAVNLIAIDSQVLCG AVKWLILEKQKPDGVFQEDAPVIHQEMTGGFRNTNEKDMALTAFVLISLQEAKEICEE QVNSLPGSITKAGDFLEANYMNLQRSYTVAIAAYALAQMGRLKGPLLNKFLTTAKDKN RWEEPGQQLYNVEATSYALLALLQLKDFDFVPPW RWLNEQRYYGGGYGSTQATEMVF QALAQYQKDVPDHKELNLDVSLQLPSRSSKI IHRIHWESASLLRSEETKENEGFTVTA EGKGQGTLSW TMYHAKAKGQLTCNKFDLKVTIKPAPETEKRPQDAKNTMILEICTRY RGDQDATMSILDISMMTGFVPDTDDLKQLANGVDRYISKYELDKAFSDRNTLI IYLDK VSHSEDDCIAFKVHQYFNVELIQPGAVKVYAYYNLAESCTRFYHPEKEDGKLNKLCRD ELCRCAEENCFIQKLDDKVTLEERLDKACEPGVDYVYKTRLVKAQLSNDFDEYIMAIE QIIKSGSDEVQVGQQRTFISPIKCREALKLEERKHYLMWGLSSDFWGEKPNLSYI IGK DTWVEHWPEEDECQDEENQKQCQDLGTFTENMW FGCPN
Cynomolgus monkey C3 amino acid sequence without predicted signal sequence (SEQ ID NO:8)
TPMYSMITPNVLRLESEETW LEAHDANGDVPVTVTVHDFPGKKLVLSSEKTVLTPAT SHMGSVTIRIPANKEFKSEKGHNKFVTVQATFGAQW EKW LVSLQSGYLFIQTDKTI YTPGSTVLCRIFTVNHKLLPVGRTVVVNIENPDGIPVKQDSLSSQNQFGILPLSWDIP ELVNMGQWKIRAYYENSPQQVFSTEFEVKEYVLPSFEVIVEPTEKFYYI YNQKGLEVT ITARFLYGKKVEGTAFVIFGIQDGEQRISLPESLKRIQIEDGSGDAVLSRKVLLDGVQ NPRPEDLVGKSLYVSVTVILHSGSDMVQAERSGIPIVTSPYQIHFTKTPKYFKPGMPF DLMVFVTNPDGSPAYRVPVAVQGEDAVQSLTQGDGVAKLS INTHPSQKPLSITVRTKK RELSEAEQATRTMEAQPYSTVGNSNNYLHLSVPRAELRPGETLNVNFLLRMDRTQEAK IRYYTYLIMNKGKLLKVGRQVREPGQDLW LPLSITTDFIPSFRLVAYYTLIGANGQR EW ADSVWVDVKDSCVGSLW KSGQSEDRQPLPGQQMTLKIEGDHGARVGLVAVDKGV FVLNKKNKLTQSKIWDW EKADIGCTPGSGKDYAGVFSDAGLTFASSSGQQTAQRAEL QCPQPAARRRRSVQLAEKRMDKVGQYPKELRKCCEHGMRENPMRFSCQRRTRYITLDE ACKKAFLDCCNYITELRRQHARASHLGLARSNLDEDI IAEENIVSRSEFPESWLWKIE ELKEAPKNGISTKLMNIFLKDSITTWEILAVSLSDKKGICVADPFEVTVMQDFFIDLR LPYSW RNEQVEIRAVLYNYRQNQELKVRVELLHNPAFCSLATAKRRHQQTVTIPPKS SLSVPYVIVPLKTGQQEVEVKAAVYHFFISDGVRKSLKW PEGIRMNKTVAVRTLDPE RLGQEGVQREDVPPADLSDQVPDTESETRILLQGTPVAQMTEDAIDAERLKHLIVTPS GCGEQNMITMTPTVIAVHYLDETEQWEKFGPEKRQGALELIKKGYTQQLAFRQPSSAF AAFLNRAPSTWLTAYW KVFSLAVNLIAIDSQVLCGAVKWLILEKQKPDGVFQEDAPV IHQEMTGGFRNTNEKDMALTAFVLISLQEAKEICEEQVNSLPGS ITKAGDFLEANYMN LQRSYTVAIAAYALAQMGRLKGPLLNKFLTTAKDKNRWEEPGQQLYNVEATSYALLAL LQLKDFDFVPPW RWLNEQRYYGGGYGSTQATEMVFQALAQYQKDVPDHKELNLDVSL QLPSRSSKIIHRIHWESASLLRSEETKENEGFTVTAEGKGQGTLSW TMYHAKAKGQL TCNKFDLKVTIKPAPETEKRPQDAKNTMILEICTRYRGDQDATMS ILDISMMTGFVPD TDDLKQLANGVDRYISKYELDKAFSDRNTLI IYLDKVSHSEDDCIAFKVHQYFNVELI QPGAVKVYAYYNLAESCTRFYHPEKEDGKLNKLCRDELCRCAEENCFIQKLDDKVTLE ERLDKACEPGVDYVYKTRLVKAQLSNDFDEYIMAIEQI IKSGSDEVQVGQQRTFISPI KCREALKLEERKHYLMWGLSSDFWGEKPNLSYI IGKDTWVEHWPEEDECQDEENQKQC QDLGTFTENMW FGCPN
38G10 Heavy chain variable region amino acid sequence (SEQ ID NO:31)
EVQLQQSGPELVKPGDSVKMSCKASGYTFTDFYMDWVKQSHGKSLEWIGYI YPHNGGT TYNQNFTGKATLTVDKSSNTAYMELHSLTSEDSAVYYCARRGGFDFDYWGQGTTLTVS S
38G10 Light chain variable region amino acid sequence (SEQ ID NO:32)
NIVMTQSPKSMSLSVGERVTLRCKASENVDTYVSWYQQKPEQSPKLLI YGASNRYTGV PDRFTGSGSATEFTLTISSVQAEDLVGYHCGQSHSYPLTFGAGTKLELK
Hz38G10 Heavy chain variable region amino acid sequence (SEQ ID NO:33)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGY IYPHNGGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 (G56A) Heavy chain variable region amino acid sequence (SEQ ID NO:34)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYI YPHNAGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 (G56T) Heavy chain variable region (SEQ ID NO:222) QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYI YPHNTGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 (N55E) Heavy chain variable region (SEQ ID NO:223) QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYI YPHEGGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 (N55Q) Heavy chain variable region (SEQ ID NO:224) QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYIYPHQGGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 Light chain variable region amino acid sequence (SEQ ID NO:35)
DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGQGTKLEIK
Hz38G10 Light chain variable region (SEQ ID NO:35) DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGQGTKLEIK
3D8 Heavy chain variable region (SEQ ID NO:77) EIQLQQSGAELVKPGASVKISCKASGYSFTGYNMNWVKQSHGKSLEWIGNINPYYGST NYNQKFKGKATLTVDKSSTTAYMQLDSLTSEDSAVYYCARGYYGGNYPFAYWGQGTLV TVSA
3D8 Light chain variable region (SEQ ID NO:78) DIQMTQSPASLSASVGETVTITCRASENIYSYLAWYQQKQGKSPQLLVYNAKTLAEGV PSRFSGSGSGTQFSLKINSLQPEDFGSYYCQHYYGTPYTFGGGSKVEIK
3G8 Heavy chain variable region (SEQ ID NO:95) EIQLQQSGAELVKPGASVKISCMASGYSFTGYNMNWVKQSHGKGLEWIGNINPYYDST SYNQKFKGKATLTVDKSSSTAYMQLNSLTSEDSAVYYCARENYDFVGFAYWGQGTLVT VSA
3G8 Light chain variable region (SEQ ID NO:96) QIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWFQQKPGSSPKPWI YVTSNLASGVP PRFSGSGSGTSYSLTISRVEAEDAATYYCQQWSTNPLTFGAGTKLELK
15C12 Heavy chain variable region (SEQ ID NO:113) EIQLQQSGAELEKPGASVKISCKASGYSFTGYNMHWVKQSHGKSLEWIGNINPYYGTT NSNQKFEDKATLTVDKSSSTAYMQLNSLTSEDSAVYYCARGI YYYGTGYPYFDFWGQG TTLTVSS
15C12 Light chain variable region (SEQ ID NO:114) DIQMTQTTSSLSASLGDRVTISCRASQDINNYLNWYLQKPDGTVKLLI YYTSRLHSGV PSRFSGSGSGTDYSLTISNLEQEDLATYFCQQGITLPWTFGGGTKLEIK
27A8 Heavy chain variable region (SEQ ID NO:131) QVQLLQPGAEFVKPGASVKLSCKASGYTFTDYWINWVKQRPGQGLEWIGNI YPGSTSA NYNEKFKSKATLTIDTSSITAYMQLSSLTSDDSAVYYCARYGYDSWFAYWGQGTLVTV SA
27A8 Light chain variable region (SEQ ID NO:132) DW LTQTPLSLPVNIGDQASISCKSTKSLLNSDGFTYLDWFLQKPGQSPHLLIYLVSN
RFSGIPDRFSGSGSETDFTLKISRVEAEDLGVYYCFQSNYLPLTFGSGTKLEIK
28C3 Heavy chain variable region (SEQ ID NO:149) QVQLQQSGPELVKPGASVKISCKASGYAFNSCWMNWVKQRPGKGLEWIGRIYPGDGDT NYNGKFKGKATLTADKSSTTAYMQLSSLTSEDSAVYFCAREGRNYGYEDYWGQGTTLT VSS
28C3 Light chain variable region (SEQ ID NO:150)
DIVLTQSPASLAVSLGQRATISCKASQSVDYDGDSYMNWYQQKPGQPPKLLIYAASDL
ESGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQANEDPRTFGGGTKLEIK
38F5 Heavy chain variable region (SEQ ID NO:167) EVMLVESGGALVKPGGSLKLSCTASGFTFSNYAMSWVRQTPEKRLEWVAQTISSGGRY TYYPDSVKGRFTISRDNARNTLYLQMSSLRSEDTAMYYCVRRYYGNSYWYFDVWGAGT TVTVSS
38F5 Light chain variable region (SEQ ID NO:168)
DIVMTQSPSSLSVSAGEKVTMNCKSSQSLLNSGNQKHYLTWYQQKPGQPPKLLIYGAS
TRGSGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDHSYPYTFGGGTKLEIK
62B11 Heavy chain variable region (SEQ ID NO:185) EVKLVESGGGLVQPGGSLKLSCAASGFTFSSYTMSWVRQTPEKRLEWVAYISSGGGTT YYPDTVKGRFTVSRDNAKNTLYLQMSSLRSEDTAMYSCARRYYRGSSLWYFDVWGAGT TVTVSS
62B11 Light chain variable region (SEQ ID NO:186)
DIVMTQSPSSLTVTAGEKVTMSCKSSQSLFNSGSQKNFLTWYQQRPGQPPKLLIYWAS
TRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLELK
62F2 Heavy chain variable region (SEQ ID NO:203) DVQLQESGPDLVKPSQSLSLTCTVTGYS ITSGYSLHWIRQFPGNKLEWMGYIHYSGST NYNPSLKSRISITRDTSKNQFFLKLNSVTSEDTATYHCARAWDYLDYWGQGTTLTVSS
62F2 Light chain variable region (SEQ ID NO:204) DIQMTQSPASLSASVGETVTITCRASENIYSQLAWYQQKQGKSPQLLVYDAKTLAEGV PSRFSGSGSDTQFSLKIISLQPEDFGRYYCHHHFGILYTFGGGTKLEMK
63A3 Heavy chain variable region (SEQ ID NO:220) DVQLQESGPGLVKPSQSLSLTCSVTGYS ITSGYYWNWIRQFPGNKLEWMGYIRYDGSN NYNPSLKNRISITRDTSKNQVFLKLNSVTPEDTATYYCARHYGYDGGAFDFWGQGTTL TVSS
63A3 Light chain variable region (SEQ ID NO:221) DIQMTQSPASLSASVGETVTITCRTSENIYNYLVWYQQKQGKSPQLLVYNAKTLEEGV PSRFSGSGSGTQFSLKVNSLQPEDFGSYYCQHHYGTPFTFGSGTKLEIK
Hz38G10 Heavy chain amino acid sequence with signal sequence underlined (SEQ ID NO:36)
MDMRVPAQLLGLLLLWLRGARCQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDW VRQAPGQRLEWMGYIYPHNGGTTYNQQFTGRVTITVDKSAS TAYMELSSLRSEDTAVY YCARRGGFDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK
Hz38G10 Heavy chain amino acid sequence without signal sequence (SEQ ID NO:37)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYI YPHNGGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ALAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV YTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK
Hz38G10 (G56A) Heavy chain amino acid sequence with signal sequence underlined (SEQ ID NO:38)
MDMRVPAQLLGLLLLWLRGARCQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDW VRQAPGQRLEWMGYIYPHNAGTTYNQQFTGRVT ITVDKSASTAYMELSSLRSEDTAVY YCARRGGFDFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTK VDKKVEPKSCDKTHTCPPCPAPALAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK
Hz38G10 (G56A) Heavy chain amino acid sequence without signal sequence (SEQ ID NO:39)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQRLEWMGYI YPHNAGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS SASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ALAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTK PREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQV
YTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL
YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Hz38G10 Light chain amino acid sequence with signal sequence underlined (SEQ ID NO:40)
MDMRVPAQLLGLLLLWLRGARCDIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWY QQKPGKAPKLLIYGASNRYTGVPSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSY PLTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASW CLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN RGEC
Hz38G10 Light chain amino acid sequence without signal sequence (SEQ ID NO:41)
DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGQGTKLEIKRTVAAPSVF IFPPSDEQLKSGTASW CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYS LSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
Human IgGl constant region (SEQ ID NO:42)
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ
SSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE
LLGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTKP
REEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Human IgGl constant region E233A/L235A (SEQ ID NO:43)
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ
SSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPA
LAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVEVHNAKTKP
REEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY
TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY
SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Human IgGl constant region L234A/L235A (SEQ ID NO:44)
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT
HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK Human IgGl constant region L234A/L235A/P329G (SEQ ID NO:45)
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF
PAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT
HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALG
APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE
SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK
Human IgGl constant region L234F/L235E/P331S (SEQ ID NO:46)
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALP ASIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK
His Tag (SEQ ID NO:47)
HHHHHH
Hz38G10 dsscFv heavy chain variable region (SEQ ID NO:225)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYI YPHNGGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 (G56A) dsscFv heavy chain variable region (SEQ ID NO:226)
QVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYI YPHNAGT TYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQGTLVTVS S
Hz38G10 dsscFv light chain variable region (SEQ ID NO:227)
DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGCGTKLEIK
Hz38G10 dsscFv (SEQ ID NO:228)
MDIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTG VPSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGCGTKLEIKGGGGSGGG GSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYI YPHNGGTTYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQ GTLVTVSS
Hz38G10 (G56A) dsscFv (SEQ ID NO:229)
MDIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTG VPSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGCGTKLEIKGGGGSGGG GSGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYI YPHNAGTTYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQ GTLVTVSS
Hz38G10 dsscFv (SEQ ID NO:230)
DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGCGTKLEIKGGGGSGGGG SGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYIY PHNGGTTYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQG TLVTVSS
Hz38G10 (G56A) dsscFv (SEQ ID NO:231)
DIQMTQSPSSLSASVGDRVTITCKASENVDTYVSWYQQKPGKAPKLLI YGASNRYTGV PSRFSGSGSGTDFTFTISSLQPEDIATYHCGQSHSYPLTFGCGTKLEIKGGGGSGGGG SGGGGSQVQLVQSGAEVKKPGASVKVSCKASGYTFTDFYMDWVRQAPGQCLEWMGYIY PHNAGTTYNQQFTGRVTITVDKSASTAYMELSSLRSEDTAVYYCARRGGFDFDYWGQG TLVTVSS

Claims

WHAT IS CLAIMED:
1. A method of treating a respiratory illness in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3), optionally wherein the respiratory illness is a severe respiratory illness.
2. A method of preventing or inhibiting the development of a respiratory illness in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3), optionally wherein the respiratory illness is a severe respiratory illness..
3. The method of claim 1 or claim 2, wherein the severe respiratory illness is selected from the group consisting of: pneumonia, acute respiratory disease (ARD), acute lung injury (ALI), acute respiratory distress syndrome (ARDS), atypical ARDS, respiratory distress syndrome (RDS), severe acute respiratory syndrome (SARS), Middle East respiratory syndrome (MERS), or coronavirus 2019 (COVID-19).
4. A method of treating acute respiratory distress syndrome (ARDS) in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
5. A method of preventing or inhibiting the development of acute respiratory distress syndrome (ARDS) in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
6. The method of any one of claims 1 to 5, wherein the human subject has a suspected or confirmed viral infection.
7. A method of inhibiting complement pathway activation in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3), wherein the human subject has a suspected or confirmed viral infection.
8. A method of treating thrombosis in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3), wherein the human subject has a suspected or confirmed viral infection.
9. A method of preventing or inhibiting the development of thrombosis in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3), wherein the human subject has a suspected or confirmed viral infection.
10. The method of claim 8 or claim 9, wherein the thrombosis is a pulmonary embolism (PE), large blood vessel blood clot, a deep vein thrombosis (DVT), or microvascular thrombosis.
11. The method of any one of claims 8 to 10, wherein the thrombosis has the potential to lead to a stroke.
12. The method of any one of claims 1 to 11, wherein the subject has symptoms of a stroke.
13. The method of any one of claims 6 to 12, wherein the viral infection is caused by a virus selected from the group consisting of: a coronavirus, an influenza virus, or a respiratory syncytial virus, optionally wherein the virus binds human ACE2 receptor.
14. The method of any one of claims 6 to 12, wherein the viral infection is caused by a coronavirus.
15. The method of claim 14, wherein the coronavirus is selected from the group consisting of: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS- CoV-1, or Middle East respiratory syndrome coronavirus (MERS-CoV).
16. The method of claim 14, wherein the coronavirus is SARS-CoV-2.
17. A method of treating a coronavirus infection in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
18. The method of claim 17, wherein the coronavirus is selected from the group consisting of: severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), SARS- CoV-1, or Middle East respiratory syndrome coronavirus (MERS-CoV).
19. The method of claim 17, wherein the coronavirus is SARS-CoV-2.
20. The method of claim 19, wherein the infection with SARS-CoV-2 results in COVID-19.
21. The method of any one of 6 to 20, wherein the viral infection is associated with a dysregulated pro-inflammatory cytokine response or cytokine storm.
22. A method of treating severe acute respiratory syndrome (SARS) in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
23. A method of preventing or inhibiting the development of severe acute respiratory syndrome (SARS) in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
24. The method of claim 22 or claim 23, wherein the human subject has a suspected or confirmed SARS-CoV-1 infection.
25. A method of treating COVID-19 in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
26. A method of preventing or inhibiting the development of COVID-19 in a human subject, the method comprising administering to the human subject a therapeutically effective amount of an antibody that specifically binds complement component C3 (C3).
27. The method of claim 25 or claim 26, wherein the human subject has a suspected or confirmed SARS-CoV-2 infection.
28. The method of any one of claims 1 to 27, wherein the human subject has lung damage, respiratory failure, kidney damage, kidney failure, liver damage, heart damage, vascular damage, thrombosis, stroke, central nervous system injury, and/or multiple organ failure.
29. The method of any one of claims 1 to 28, wherein the human subject has one or more symptoms selected from the group consisting of: hypoxemia, cough, wheezing, dyspnea, hyperpnea, pulmonary /lung inflammation, shortness of breath, labored breathing, rapid breathing, accumulation of alveolar fluid, pulmonary edema, vascular leakage, lymphocyte infiltration in the lung, lymphopenia, fever, chills, shaking chills, increased heart rate, chest pain, low blood pressure, headache, confusion, seizures, extreme tiredness, sepsis, bluish coloring of nails or lips, toe rashes/redness, toe swelling, loss of sense of smell, loss of sense of taste, and diarrhea.
30. The method of claim 29, wherein the human subject has mild, moderate, or severe hypoxemia as determined by Partial Pressure of arterial oxygen/Fraction of inspired oxygen (PaCk/FiCk) or positive end-expiratory pressure (PEEP).
31. The method of claim 30, wherein the human subject has severe hypoxemia with a PaCk/FiCk of less than 100.
32. The method of any one of claims 1 to 31, wherein the antibody has at least one or more of the following properties:
(a) is an antagonist of C3;
(b) inhibits C3 activity;
(c) inhibits C3 cleavage;
(d) inhibits C3 cleavage and C3a release;
(e) inhibits complement activation;
(f) inhibits activation of the alternative complement pathway;
(g) inhibits activation of the classical complement pathway; and/or
(h) inhibits activation of the alternative complement pathway and classical complement pathway.
33. The method of any one of claims 1 to 32, wherein the antibody comprises: (1) a heavy chain variable region (VH) comprising VH complementarity determining region (CDR)1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:31, and a light chain variable region (VL) comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:32;
(2) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:33, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35;
(3) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:34, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35;
(4) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:222, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35;
(5) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:223, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35;
(6) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:224, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:35;
(7) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:77, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:78;
(8) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:95, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 96;
(9) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 113, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 114; (10) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 131, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 132;
(11) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 149, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 150;
(12) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 167, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 168;
(13) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO: 185, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO: 186;
(14) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:203, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:204; or
(15) a VH comprising VH CDR1, VH CDR2, and VH CDR3 from the amino acid sequence set forth in SEQ ID NO:220, and a VL comprising VL CDR1, VL CDR2, and VL CDR3 from the amino acid sequence set forth in SEQ ID NO:221.
34. The method of any one of claim 33, wherein the antibody comprises:
(a) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 16, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 10, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:20, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (b) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:25, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 16, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 17, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:25, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:26, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24;
(c) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:27, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:28, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:29, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:27, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:30, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24;
(d) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:48, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:49, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:50, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:48, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:51, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24; (e) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:52, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:53, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:54, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:52, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:55, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24;
(f) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:56, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 15, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:57, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:9, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:58, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 18, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:56, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 13, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 14; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 19, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:59, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:21, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:22, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:23, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:24;
(g) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:60, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:64, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 75;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:61, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:65, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 75;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:60, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:67, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 75;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:62, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:64, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 69, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:71, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:73, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 75; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:63, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:68, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:70, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:72, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:74, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 76; (h) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:79, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:83, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 93;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:80, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:84, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 93;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:79, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:85, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 93;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:81, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:83, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 87, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:89, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:91, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 93; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:82, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:86, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:88, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:90, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:92, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 94;
(i) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:97, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 111;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:98, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 102, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 111;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:97, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 103, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 111;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:99, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 105, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 107, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 109, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO : 111 ; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 100, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 104, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 106, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 108, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:l 10, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 112;
(j) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 115, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 120, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 115, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 121, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 117, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 123, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 125, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 127, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 118, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 122, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 124, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 126, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 130; (k) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 133, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 138, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 133, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 139, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 135, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 141, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 143, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 145, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 136, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 140, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 142, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 144, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 146, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 148;
(1) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:151, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 156, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:151, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 157, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 153, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 161, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 163, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 154, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 158, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 160, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 162, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 164, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 166;
(m) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 169, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 173, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 170, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 174, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 169, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 175, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 181, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 171, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 173, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 179, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 171, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 183; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 172, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 176, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 178, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 180, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 182, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 184; (n) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 187, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 191, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 188, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 192, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 187, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 193, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 189, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 191, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 195, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 197, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:201; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 190, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 194, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:200, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:202; or
(o) (i) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:205, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:209, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218;
(ii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:206, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:210, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218;
(iii) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:205, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:211, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218;
(iv) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:207, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:209, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO:213, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:232, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:216, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:218; or
(v) a VH comprising a VH CDR1 comprising the amino acid sequence set forth in SEQ ID NO:208, a VH CDR2 comprising the amino acid sequence set forth in SEQ ID NO:212, and a VH CDR3 comprising the amino acid sequence set forth in SEQ ID NO :214, and a VL comprising a VL CDR1 comprising the amino acid sequence set forth in SEQ ID NO:215, a VL CDR2 comprising the amino acid sequence set forth in SEQ ID NO:217, and a VL CDR3 comprising the amino acid sequence set forth in SEQ ID NO:219.
35. The method of any one of claims 1 to 32, wherein the antibody comprises:
(a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHN GGTT YN QNF T G (SEQ ID NO: 10), YIYPHNGGTTYNQQFTG (SEQ ID NO:25), or YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11); and/or
(b) a light chain variable region comprising a light chain variable region
CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
36. The method of any one of claims 1 to 32, wherein the antibody comprises:
(a) a heavy chain variable region comprising a heavy chain variable region CDR1 comprising the amino acid sequence GYTFTDFYMD (SEQ ID NO:9), a heavy chain variable region CDR2 comprising the amino acid sequence YIYPHNAGTTYNQQFTG (SEQ ID NO:27), and a heavy chain variable region CDR3 comprising the amino acid sequence RGGFDFDY (SEQ ID NO: 11), and
(b) a light chain variable region comprising a light chain variable region CDR1 comprising the amino acid sequence KASENVDTYVS (SEQ ID NO: 12), a light chain variable region CDR2 comprising the amino acid sequence GASNRYT (SEQ ID NO: 13), and a light chain variable region CDR3 comprising the amino acid sequence GQSHSYPLT (SEQ ID NO: 14).
37. The method of claim 35 or claim 36, wherein (a) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or (b) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35.
38. The method of claim 35 or claim 36, wherein (a) the heavy chain variable region comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:33 or SEQ ID NO:34; and/or (b) the light chain variable region comprises a sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:35.
39. The method of claim 38, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35.
40. The method of claim 38, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35.
41. The method of claim 33 or 34, wherein
(a) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31;
(b) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:33;
(c) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:34;
(d) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222;
(e) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223;
(f) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224;
(g) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77; (h) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:95;
(i) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:l 13;
(j) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131 ;
(k) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149;
(l) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167;
(m) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185;
(n) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203;
(o) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220;
(p) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32;
(q) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(r) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78;
(s) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:96;
(t) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:l 14;
(u) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132;
(v) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150; (w) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168;
(x) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186;
(y) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204;
(z) the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221;
(aa) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:31 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:32;
(bb) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(cc) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(dd) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(ee) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(fif) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:35;
(gg) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:78;
(hh) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 95 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:96;
(ii) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:l 13 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 114;
(jj) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 132;
(kk) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 150;
(11) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 168;
(mm) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 186; (nn) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:204; or
(oo) the heavy chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises a sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:221.
42. The method of claim 33 or 34, wherein
(a) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31;
(b) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33;
(c) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:34;
(d) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222;
(e) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223;
(f) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224;
(g) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77;
(h) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:95;
(i) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:l 13;
(j) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131 ; (k) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149;
(l) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167;
(m) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185;
(n) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203;
(o) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220;
(p) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32;
(q) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(r) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78;
(s) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:96;
(t) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:l 14;
(u) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132;
(v) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150;
(w) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168;
(x) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186;
(y) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204; (z) the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221;
(aa) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:31 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:32;
(bb) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(cc) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(dd) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(ee) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(fif) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:35;
(gg) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:78; (hh) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 95 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:96;
(ii) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:l 13 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 114;
(jj) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 132;
(kk) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150;
(11) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 168;
(mm) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 186;
(nn) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:204; or
(oo) the heavy chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises a sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:221.
43. The method of claim 33 or 34, wherein
(a) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:31;
(b) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33;
(c) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34;
(d) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:222;
(e) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:223;
(f) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:224;
(g) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:77;
(h) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:95;
(i) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:113;
(j) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:131;
(k) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 149;
(l) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 167;
(m) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:185; (n) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:203;
(o) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:220;
(p) the light chain variable region comprises amino acid sequence of SEQ ID NO:32;
(q) the light chain variable region comprises amino acid sequence of SEQ ID NO:35;
(r) the light chain variable region comprises amino acid sequence of SEQ ID NO:78;
(s) the light chain variable region comprises amino acid sequence of SEQ ID NO:96;
(t) the light chain variable region comprises amino acid sequence of SEQ ID NO: 114;
(u) the light chain variable region comprises amino acid sequence of SEQ ID NO: 132;
(v) the light chain variable region comprises amino acid sequence of SEQ ID NO: 150;
(w) the light chain variable region comprises amino acid sequence of SEQ ID NO: 168;
(x) the light chain variable region comprises amino acid sequence of SEQ ID NO: 186;
(y) the light chain variable region comprises amino acid sequence of SEQ ID NO:204;
(z) the light chain variable region comprises amino acid sequence of SEQ ID NO:221; (aa) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:31 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:32;
(bb) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:33 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35;
(cc) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:34 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35;
(dd) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:222 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35;
(ee) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:223 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35; (ff) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:224 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:35;
(gg) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:77 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:78;
(hh) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:95 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 96;
(ii) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 113 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 114;
(jj) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 131 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 132;
(kk) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 149 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 150;
(11) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 167 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 168;
(mm) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 185 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 186;
(nn) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:203 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:204; or
(oo) the heavy chain variable region comprises the amino acid sequence of SEQ ID NO:220 and the light chain variable region comprises the amino acid sequence of SEQ ID NO:221.
44. The method of any one of claims 1 to 43, wherein the antibody is a monoclonal antibody.
45. The method of any one of claims 1 to 44, wherein the antibody is a humanized antibody.
46. The method of any one of claims 1 to 45, wherein the antibody is a bispecific antibody or a multispecific antibody.
47. The method of any one of claims 1 to 46 wherein the antibody is an antibody fragment comprising at least one antigen-binding site.
48. The method of claim 47, wherein the antibody fragment is a Fab, Fab', F(ab')2, Fv, scFv, (SCFV)2, single chain antibody, dual variable region antibody, diabody, or nanobody.
49. The method of any one of claims 1 to 46, wherein the antibody is an IgGl antibody.
50. The method of any one of claims 1 to 46, wherein the antibody is an IgG2 antibody.
51. The method of any one of claims 1 to 46, wherein the antibody is an IgG4 antibody.
52. The method of any one of claims 1 to 32, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:37 and a light chain comprising the amino acid sequence of SEQ ID NO:41.
53. The method of any one of claims 1 to 32, wherein the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:39 and a light chain comprising the amino acid sequence of SEQ ID NO:41.
54. The method of any one of claims 1 to 53, wherein the antibody is attached to a half-life extending moiety.
55. The method of any one of claims 1 to 54, which comprises administering the antibody as part of a combination therapy.
56. The method of claim 55, wherein the combination therapy includes use of a medical device.
57. The method of claim 56, wherein the medical device is a ventilator or an ECMO machine.
58. The method of any one of claims 1 to 57, which comprises administering at least one additional therapeutic agent.
PCT/US2021/031268 2020-05-08 2021-05-07 Therapeutic uses of c3-binding agents WO2021226442A2 (en)

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