RU2808018C2 - Antibodies to surface determinants of s. aureus - Google Patents

Abstract

FIELD: biochemistry.

SUBSTANCE: invention is related a pharmaceutical composition for use in the treatment of S. aureus infection. A method for treating an S. aureus infection in a patient, a method for reducing bacterial agglutination of S. aureus and/or the formation of a thromboembolic lesion in a patient, a method for delaying the onset of sepsis, a method for preventing or reducing the severity of sepsis associated with S. aureus using the specified composition are also disclosed.

EFFECT: invention has the ability to effectively treat S. aureus infection.

18 cl, 18 dwg, 5 tbl, 11 ex

Description

[0001] The present disclosure generally relates to antibodies that bind to surface determinants of Staphylococcus aureus (S. aureus) and antibodies that bind to secreted toxins of S. aureus. The present disclosure also relates to combinations of antibodies that bind to surface determinants of S. aureus, together with antibodies that bind to secreted toxins of S. aureus, compositions containing such combinations of antibodies, and methods of preventing S. aureus-associated diseases, including the administration of such combinations of antibodies.

[0002] Staphylococcus aureus is a Gram-positive, aerobic, aggregate-forming coccal bacterium that commonly colonizes the nose and skin of healthy individuals. Approximately 20-30% of the population is colonized with S. aureus at any time. The bacteria Staphylococcus aureus, which is also sometimes called “staphylococcus”, “Staph. aureus" or "S. aureus" are considered opportunistic pathogens that cause secondary infections (eg, pustules, boils, and other soft tissue infections) and systemic infections (eg, pneumonia, sepsis, osteomyelitis, and endocarditis).

[0003] Mucosal and epidermal barriers (skin) normally protect against S. aureus infections. Disruption of these natural barriers due to injury (eg, burns, trauma, and surgery) greatly increases the risk of infection. Diseases that compromise the immune system (such as diabetes, end-stage renal disease, and cancer) also increase the risk of infection. Infections caused by opportunistic S. aureus can become severe, causing a number of diseases or conditions, non-limiting examples of which include bacteremia, cellulitis, eyelid infections, food poisoning, joint infections, skin infections, toxic epidermal necrolysis, toxic shock syndrome, pneumonia, osteomyelitis, endocarditis, meningitis and abscess formation.

[0004] S. aureus expresses a number of surface determinant antigens, including the serine-aspartic acid repeat proteins SdrC, SdrD and SdrE, the agglutinin proteins ClfA and ClfB, and the iron-regulated surface determinant proteins IsdA, IsdB, IsdC, IsdE and IsdH , protein A (SpA) of S. aureus and the polysaccharide poly-N-acetylglucosamine (PNAG). These surface antigens play a role in host tissue colonization, evasion of the host immune response, and bacterial fitness. Mutations in ClfA, SpA, IsdA, IsdB, and IsdH have been shown to reduce the virulence of S. aureus.

[0005] Proteins such as IsdH play a role in the ability of S. aureus to evade certain host immune responses, such as neutrophil-mediated phagocytosis, a process that is critical for the initiation of S. aureus infection. IsdH is part of a complex that is activated under iron-limited conditions, serving to bind hemoglobin and the haptoglobin-hemoglobin complex, and then extract and transport heme into the cytoplasm. IsdH contains three N-terminal NEAr Transporter (NEAT) motifs, and the determined structure of NEAT1 indicates that some residues in this motif are involved in ligand binding. IsdH-defective mutants of S. aureus have been shown to have reduced virulence compared to the wild type and are more rapidly taken up by human neutrophils in the presence of serum opsonins. The protective mechanism of IsdH appears to be due to the accelerated degradation of opsonin C3b in the blood serum. Thus, IsdH plays a role in the antiphagocytic properties of S. aureus.

[0006] ClfA is a virulence factor that binds fibrinogen. This function of ClfA results in an additional contribution to the antiphagocytic properties of S. aureus. In addition, ClfA also promotes the agglutination of S. aureus in the blood and the formation of biofilm on biomaterial surfaces.

[0007] S. aureus also expresses several additional virulence factors, including capsular polysaccharides and protein toxins. One virulence factor often associated with S. aureus infection is alpha-toxin (also known as alpha-hemolysin or Hla), a pore-forming and hemolytic exoprotein produced by the most pathogenic strains of S. aureus. The toxin forms heptameric pores in the membranes of sensitive cells such as leukocytes, platelets, erythrocytes, peripheral blood monocytes, macrophages, keratinocytes, fibroblasts and endothelial cells. The formation of alpha-toxin pores often leads to cell dysfunction or lysis. It can also lead to disruption of tight junctions in the epithelium and endothelium and dysregulation of the immune system.

[0008] S. aureus is currently the leading cause of infectious disease-related mortality in the United States and a leading cause of nosocomial infection. In addition, the increasing resistance of S. aureus to antibiotics has become a problem. Thus, it would be desirable to develop effective alternative methods for diagnosing and treating S. aureus infections, including combination antibody therapies.

[0009] As previously disclosed in US Provisional Application No. 61/440581 and International Application No. PCT/US2012/024201 (published as WO2012/109205), the contents of each of which are incorporated herein by reference, it has been shown that antibodies that bind to S. aureus alpha-toxin reduce the severity of CA-MRSA disease in a mouse model of dermatotonecrosis and promote bacterial clearance in a mouse model of staphylococcal pneumonia. Thus, such antibodies can be used to treat various S. aureus-associated diseases.

[0010] In addition to antibodies that bind to S. aureus alpha toxin, the present disclosure provides antibodies directed against S. aureus surface determinant antigens, as well as combinations thereof. The present invention provides compositions containing such antibodies, or combinations of such antibodies, as well as methods of preventing and/or treating S. aureus-associated diseases using such antibodies or combinations of such antibodies. Methods for preventing and/or treating S. aureus-related diseases using antibodies that bind to S. aureus alpha-toxin are described in US Provisional Patent Application No. 61/440581 and International Application No. PCT/US2012/024201 (published as WO2012/109205), the contents of which are incorporated herein by reference, as well as in the US provisional patent application filed concurrently with this application by Sellman et al., entitled “Methods for the Treatment of S. Aureus-Related Diseases.” of Treating S. Aureus Associated Diseases”), the contents of which are incorporated herein by reference.

[0011] The present invention also provides certain combinations of antibodies, such as an antibody that binds to S. aureus surface determinant in combination with an antibody that binds to S. aureus alpha toxin, wherein such combinations together act synergistically. The present invention also relates to combining an antibody that targets an S. aureus surface determinant antigen involved in evading host opsonophagocytic functions with an antibody that targets a S. aureus secreted toxin involved in direct damage to host cells.

DESCRIPTION OF GRAPHIC MATERIALS

[0012] In FIG. Figure 1 shows the percentage of opsonophagocytic killing (OPK) induced by IsdH antibodies B11, 2F4, and A7 compared with the percentage of OPK induced by the control antibody R347 when tested with S. aureus strains Newman and USA300.

[0013] Figure 2 shows the binding of anti-IsdH antibodies B11, 2F4, A7 and 1C1, compared to the control antibody R347, to S. aureus strains ARC2081 and USA300.

[0014] Figure 3A is a plot of competitive binding between antibody 1C1 and haptoglobin (Hp) for binding to the Neat-1 subunit on IsdH versus competitive binding between control antibody R347 and Hp. Figure 3B is a plot of competitive binding between antibody 2F4 and Hp for binding to the Neat-2 subunit on IsdH versus competitive binding between control antibody R347 and Hp.

[0015] Figure 4A shows the concentration of colony forming units (CFU) of S. aureus measured in a mouse model of bacteremia in the presence of antibody 1C1. CFU concentration is given as log ₁₀ [CFU/ml]. Figure 4B shows the concentration of S. aureus CFU measured in a mouse model of bacteremia in the presence of antibody A7. CFU concentration is given as log ₁₀ [CFU/ml]. Figure 4C shows the concentration of S. aureus CFU measured in a mouse model of bacteremia in the presence of the IsdH0016 antibody. CFU concentration is given as log ₁₀ [CFU/ml]. Figure 4D shows the concentration of S. aureus CFU measured in a mouse model of bacteremia in the presence of the IsdH003 antibody. CFU concentration is given as log[CFU/ml].

[0016] Figure 5 shows the concentration of colony forming units (CFU) of S. aureus measured in a mouse model of bacteremia in the presence of antibody 2F4. CFU concentration is given as log ₁₀ [CFU/ml].

[0017] Figure 6 illustrates 2F4 binding, measured as the percentage of maximum binding compared to the control antibody R347, at time points _T0 , _T1h and _T4h in strains ARC2379 (USA100), ARC2081 (USA200) and BAA-1556 ( USA 300) S. aureus.

[0018] Figure 7A shows the percentage of OPK induced by antibody 2F4 compared to the percentage of OPK induced by control antibody R347 when tested with S. aureus strain Newman. Figure 7B shows the percentage of OPK induced by antibody 2F4 compared to the percentage of OPK induced by control antibody R347 when tested with S. aureus strain ARC634. Figure 7C shows the percentage of OPK induced by antibody 2F4 compared to the percentage of OPK induced by control antibody R347 when tested with S. aureus strain ARC2081 (USA200). Figure 7D shows the percentage of OPK induced by antibody 2F4 compared to the percentage of OPK induced by control antibody R347 when tested with S. aureus strain BAA-1556 (USA300).

[0019] Figure 8 shows the increase in 2F4 affinity for IsdH and the Neat-2 subunit in the hulgGFc uptake assay.

[0020] Figure 9A shows the kidney CFU concentration for S. aureus strain USA300 (administered at an initial concentration of 2.05e8) measured in an organ loading model after treatment with 2F4 alone, 2A3 alone, or a combination of 2F4 and 2A3. CFU concentrations are reported as log ₁₀ [CFU/organ]. Figure 9B shows the kidney CFU concentration for S. aureus strain USA300 (administered at an initial concentration of 2.05e8) measured in an organ loading model after treatment with 2F4 alone, 2A3 alone, or a combination of 2F4 and 2A3. CFU concentrations are reported as log ₁₀ [CFU/organ].

[0021] Figure 10 shows that antibodies to ClfA inhibit the binding of ClfA to immobilized fibrinogen. Antibodies 23D6, 27H4, 11H10 to ClfA showed increased inhibition of fibrinogen binding compared to the control R347.

[0022] Figure 11 shows that antibodies to ClfA inhibit the agglutination of S. aureus in human plasma. The ability of anti-ClfA antibodies 23D6, 27H4, 11H10 to inhibit agglutination of three different S. aureus strains (NR S112, BAA 1556 and UAMS-1) was tested compared to the control R347, in the absence of mAb and ClfA protein. In this analysis, anti-ClfA antibody 11H10 had the highest strain coverage.

[0023] Figure 12 shows that anti-ClfA antibody 11H10 binds to a different epitope on ClfA compared to anti-ClfA antibodies 23D6 and 27H4.

[0024] Figure 13 demonstrates that anti-ClfA antibody 11H10 and anti-AT antibody LC10 reduce the bacterial load in the heart.

[0025] Figure 14 shows the effect of anti-ClfA antibody 11H10 in a mouse model of sepsis.

[0026] Figure 15 shows the effect of anti-ClfA antibody 11H10, anti-AT antibody LC10, and the combination of anti-ClfA antibody 11H10 with anti-AT antibody LC10 in a murine model of sepsis (grade IV, lethal infection) resulting from infection with CA-MRSA USA300.

[0027] Figure 16 shows the effect of combining anti-ClfA antibody 11H10 with anti-AT antibody LC10 in a mouse model of sepsis (grade IV, lethal infection) resulting from infection with HA-MRSA USA100.

[0028] Figure 17 shows the effect of combining anti-ClfA antibody 11H10 with anti-AT antibody LC10 in a mouse model of sepsis (stage IV, lethal infection) resulting from infection with HA-MRSA USA200.

[0029] Figure 18 shows the effect of combining anti-IsdH antibody 2F4 with anti-AT antibody LC10 in a mouse model of sepsis (stage IV, lethal infection) resulting from infection with HA-MRSA USA100.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0030] We now turn in detail to some illustrative embodiments of the present disclosure, some examples of which are illustrated in the accompanying drawings.

[0031] Disclosed herein are antibodies, including human, humanized and/or chimeric forms, as well as fragments, derivatives/conjugates and compositions thereof, that bind to S. aureus surface determinant antigens and antibodies that bind to secreted toxins from S. aureus. Such antibodies may be useful for detecting and/or visualizing S. aureus and thus may be useful in diagnostic methods and assays. The antibodies described herein also target surface determinants of S. aureus, thereby affecting colonization and immunosurveillance, making these antibodies suitable for therapeutic and prophylactic applications. Likewise, the antibodies described herein can bind secreted toxins from S. aureus, thereby reducing the virulence of S. aureus infection. Combining antibodies that target both surface determinants and secreted toxins of S. aureus may enhance the therapeutic and preventive effects achieved by any antibody when administered individually.

[0032] S. aureus expresses a number of surface determinant antigens that are important for S. aureus colonization, immune evasion, and fitness. Such surface determinants include SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH, SpA, FnbA and PNAG. The antibodies disclosed herein can be directed to antigens that are surface determinants.

[0033] S. aureus also produces a large number of secreted and cell-associated proteins, many of which are involved in pathogenesis, such as alpha toxin (AT), beta toxin, gamma toxin, delta toxin, leukocidin, toxic syndrome toxin shock shock (TSST), enterotoxins, coagulase, protein A, fibrinogen, etc. Alpha toxin is one of the virulence factors of Staphylococcus aureus and is produced by most pathogenic strains of S. aureus.

A. Antibodies directed against surface determinants and secreted toxins of S. aureus

[0034] As used herein, the terms “antibody,” “antibodies” (also known as immunoglobulins), and “antigen-binding fragments” include monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies formed from at least two different epitope binding fragments (eg, bispecific antibodies), human antibodies, humanized antibodies, camel antibodies, chimeric antibodies, single chain Fv (scFv), single chain antibodies, single domain antibodies, domain antibodies, Fab fragments, F(ab')2 fragments, fragments antibodies that exhibit the desired biological activity (eg, antigen-binding moiety), disulfide-linked Fv (dsFv) and anti-idiotypic (anti-Id) antibodies (including, for example, anti-Id antibodies to the antibodies provided herein), intracellular antibodies, and epitope-binding fragments of any of the above. In particular, antibodies include immunoglobulin molecules and immunologically active fragments of immunoglobulin molecules, ie, molecules that contain at least one antigen-binding site. Immunoglobulin molecules can be of any isotype (e.g. IgG, IgE, IgM, IgD, IgA and IgY), subisotype (e.g. IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or allotype (e.g. Gm, e.g. G1m(f , z, a, or x), G2m(n), G3m(g, b, or c), Am, Em, and Km(1, 2, or 3)). The antibodies can be obtained from any mammal, including, without limitation, humans, monkeys, pigs, horses, rabbits, dogs, cats, mice and the like, or other animals such as birds (eg, chickens).

[0035] In certain embodiments, an antibody or immunospecific fragment thereof of the present invention includes an antigen binding domain. The antigen binding domain is formed by the variable regions of an antibody, which differ among different antibodies. Naturally occurring antibodies contain at least two antigen-binding domains, i.e. they are at least bivalent. As used herein, the expression “antigen-binding domain” includes a region that specifically binds to an epitope of an antigen (eg, a cell surface antigen or a soluble antigen). The antigen binding domain of an antibody typically includes at least a portion of an immunoglobulin heavy chain variable region and at least a portion of an immunoglobulin light chain variable region. The binding site formed by these variable regions determines the specificity of the antibody.

[0036] As used herein, unless specifically stated otherwise, “mutation” includes the addition, deletion, substitution (including conservative substitution) or other change of at least one amino acid or nucleic acid. A “conservative substitution,” unless specifically stated otherwise, refers to the replacement of a first amino acid with a second amino acid that does not substantially change the chemical, physical, and/or functional properties of the antibody or antigen-binding fragment thereof (e.g., the antibody or antigen-binding fragment thereof retains the same charge, structure , polarity, hydrophobicity/hydrophilicity and/or retains functions such as the ability to bind alpha toxin and thus reduce the virulence of S. aureus). Conservative substitution also refers to the replacement of a first nucleic acid with a second nucleic acid encoding a previously described conservative amino acid substitution.

[0037] Antibodies provided herein include full-length or intact antibodies, antibody fragments, native sequence antibodies or amino acid variants of native antibodies, human, humanized, post-translationally modified antibodies, chimeric or hybrid antibodies, immunoconjugates, and functional fragments thereof . Antibodies may be modified in the Fc region and some modifications may provide desired effector functions or altered serum half-life.

[0038] Amino acid numbering in the variable domain, complementarity determining regions (CDRs), and framework regions (FRs) of an antibody is as defined by Kabat, unless otherwise noted, as set forth in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. . Public Health Service, National Institutes of Health, Bethesda, Maryland. Public Health Service, National Institutes of Health, Bethesda, Maryland (1991). When using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to the truncation or insertion into the FR or CDR of the variable domain. Kabat residue numbering can be determined for a given antibody by aligning regions of the antibody with sequence homology to the antibody with a “standard” Kabat numbered sequence. Maximum alignment of framework residues often requires the insertion of “spacer” residues into the numbering system to be used for the Fv region. In addition, the identity of certain individual residues in any specified Kabat numbered region may vary from antibody chain to antibody chain due to interspecies or allelic divergence.

[0039] In certain embodiments, isolated antibodies are provided. As used herein, the term “isolated antibody” refers to an antibody that is substantially free of other antibodies and molecules normally present in the natural cellular environment. Thus, in some embodiments, the antibodies provided are isolated antibodies and have been separated from antibodies with different antigen specificities. The isolated antibody may be a monoclonal antibody or a polyclonal antibody. An isolated antibody that specifically binds to an epitope, isoform, or variant of a surface antigen or secreted toxin of S. aureus may nevertheless be cross-reactive with other related antigens, for example, from other species (eg, homologue species of Staphylococcus). The isolated antibody provided may be substantially free of one or more other cellular materials. In some embodiments, a combination of “isolated” monoclonal antibodies is provided and refers to antibodies having different specificities and combined into a specific composition.

[0040] Also disclosed are isolated nucleic acid sequences that encode the amino acid sequences of the disclosed antibodies and antigen binding fragments thereof. Due to the degeneracy of the nucleotide code, more than one nucleotide can be present at any position in the nucleic acid while still coding for the same amino acid. In some embodiments, nucleic acid sequences are provided that encode amino acid sequences that are 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to (or any percentage therebetween) the amino acid sequences of a disclosed antibody or antigen-binding fragment thereof that bind to a surface antigen or secreted toxin of S. aureus. In additional embodiments, the nucleic acid sequences encode amino acid sequences that retain the functional abilities of the disclosed antibodies and antigen-binding fragments thereof, such as binding a surface antigen or secreted toxin of S. aureus and thereby reducing colonial growth, evasion of opsonophagocytosis, or toxicity of the secreted S. aureus toxin. aureus.

[0041] In various embodiments, antibodies or fragments disclosed herein can specifically bind a S. aureus surface antigen or secreted toxin polypeptide or an antigenic fragment thereof. In certain embodiments, the surface antigen is SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH, SpA, FnbA, or PNAG. In additional embodiments, the surface antigen is IsdH. In other embodiments, the surface antigen is ClfA. In some embodiments, the secreted toxin is alpha toxin or phenol-soluble modulin. In additional embodiments, the secreted toxin is an alpha toxin. Certain amino acid and nucleic acid sequences for anti-alpha toxin antibodies useful in the present disclosure are disclosed in US Provisional Patent Application No. 61/440581 and International Application No. PCT/US2012/024201 (published as WO2012/109205), the contents of each of which are thus included in their entirety.

[0042] Antibodies provided herein may specifically bind to one or more epitopes specific for a surface determinant antigen protein or secreted toxin of S. aureus, and generally do not specifically bind to other polypeptides. As used herein, the term “epitope” refers to a peptide, subunit, fragment, portion, oligomer, or combination thereof that can be bound by an antibody.

[0043] In certain embodiments, an antibody to a surface determinant antigen or secreted toxin of S. aureus or an antigen-binding fragment thereof may bind a species-conserved epitope. In some embodiments, the antibody or antigen-binding fragment thereof binds a surface determinant antigen or secreted toxin of S. aureus or a homolog or ortholog of another bacterial species, as well as antigenic fragments thereof. In some embodiments, an antibody or an antigen-binding fragment thereof can bind to one or more isoforms of a surface determinant antigen or a secreted toxin.

[0044] In various embodiments, the epitope includes at least a portion of an antigen that is a surface determinant of S. aureus. These surface determinant antigens may include SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH, SpA, FnbA or PNAG. In some embodiments, the antigen is IsdH. In other embodiments, the antigen is ClfA. In some embodiments, the epitope includes at least a portion of a secreted S. aureus toxin. In some embodiments, the secreted toxin is an alpha toxin that is involved in the formation of the alpha toxin heptameric complex.

[0045] The epitope may comprise any amino acid sequence containing at least 3 contiguous amino acid residues from the amino acid sequence of the target antigen. An epitope may contain longer amino acid sequences, up to and including the entire amino acid sequence of the target antigen. In some embodiments, the epitope contains at least 4 amino acid residues, at least 5 amino acid residues, at least 6 amino acid residues, at least 7 amino acid residues, at least 8 amino acid residues, at least 9 amino acid residues, at least at least 10 amino acid residues, at least 11 amino acid residues, at least 12 amino acid residues, at least 13 amino acid residues, at least 14 amino acid residues, or at least 15 amino acid residues from the amino acid sequence of the target antigen. In some other embodiments, the epitope comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 contiguous or non-contiguous amino acid residues from the amino acid sequence of the target antigen.

[0046] In certain embodiments, combinations are provided comprising an isolated antibody or antigen-binding fragment thereof that specifically binds to a secreted S. aureus toxin and an isolated antibody that specifically binds to an S. aureus surface determinant antigen. In additional embodiments, an antibody that binds an S. aureus surface determinant antigen binds an antigen selected from SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH, SpA, FnbA, or PNAG. In yet other embodiments, the surface determinant antigen is IsdH. In certain embodiments, an antibody that binds a secreted toxin from S. aureus binds a toxin selected from alpha toxin and phenol-soluble modulin. In additional embodiments, the secreted toxin is an alpha toxin.

[0047] In certain embodiments, the antibody or combination of antibodies is present in an aqueous solution. In other embodiments, the antibody or combination of antibodies is present in powder or lyophilized form. In certain embodiments, the antibody or combination of antibodies is present in a concentration sufficient for therapeutic or diagnostic applications. In some embodiments, the antibody or combination of antibodies is present in a sterile vessel or container.

[0048] In certain embodiments, an antibody or antigen-binding fragment thereof that can bind a S. aureus surface antigen or secreted toxin is derived from a parent antibody. As used herein, the term “parent antibody” refers to an antibody that is encoded by the amino acid sequence used to produce the variant or derivative antibody described herein. The original polypeptide may contain a native antibody sequence (i.e., a naturally occurring antibody polypeptide, including a naturally occurring allelic variant) or an antibody sequence with pre-existing amino acid sequence modifications (such as insertions, deletions, and/or substitutions) in the naturally occurring the nature of sequence. The parent antibody may be a humanized antibody or a human antibody. In specific embodiments, antibodies to a surface antigen or secreted toxin of S. aureus and antigen-binding fragments thereof are variants of the parent antibody. As used herein, the expression “variant” refers to an antibody or antigen binding fragment thereof that differs in its amino acid sequence from the amino acid sequence of the “parent” antibody or antigen binding fragment thereof due to the addition, deletion and/or substitution of one or more amino acid residues from the sequence of the parent antibody .

[0049] Present antibodies to a surface antigen or secreted toxin of S. aureus and antigen binding fragments thereof contain at least one antigen binding domain. The antigen-binding portion of an antibody contains one or more antibody fragments that retain the ability to specifically bind an antigen. These retained portions may comprise a heavy and/or light chain variable region from the parent antibody or a variant of the parent antibody.

B. Antibodies to IsdH

[0050] As used herein, the expressions “percentage of sequence identity” or “homology” are defined as the percentage of amino acid residues or nucleotides in a candidate sequence that are identical to amino acid residues or nucleotides in reference sequences after sequence alignment and introduction of gaps, if necessary. , to achieve the maximum percentage of sequence identity, and excluding conservative substitutions. Optimal sequence alignment for comparison can be obtained, in addition to the manual method, using local homology search algorithms known in the art, or using computer programs that use these algorithms (for example, GAP, BESTFIT, FASTA, BLAST P, BLAST N and TFASTA).

[0051] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to an IsdH surface antigen comprises a heavy chain variable region (VH) that is 65%, 70%, 75%, 80%, 85%, 90% , 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 80, 82, 84, 86, or 88. In certain embodiments, an antibody or antigen binding fragment thereof that specifically binds IsdH comprises a light chain variable region (VL) that is 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 81, 83, 85, 87, or 89. In certain embodiments, an antibody or antigen-binding fragment thereof that specifically binds IsdH contains a VH that is 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 80, 82, 84, 86 or 88 and VL containing the amino acid sequence of SEQ ID NO: 81, 83, 85, 87 or 89.

[0052] In specific embodiments, an antibody or antigen binding fragment thereof that specifically binds IsdH comprises VH and VL, wherein VH and VL are selected from the group consisting of SEQ ID NOs: 80 and 81; SEQ ID NO: 82 and 83; SEQ ID NO: 84 and 85; SEQ ID NOs: 86 and 87 and SEQ ID NOs: 88 and 89. In certain embodiments, an antibody or antigen binding fragment thereof that specifically binds IsdH comprises VH and VL, wherein VH and VL correspond to SEQ ID NOs: 80 and 81 Example 7, Table 12, provides exemplary VH and VL sequences provided herein that may be present in any combination to form an antibody to a surface antigen or an antigen binding fragment thereof.

[0053] In additional embodiments, an antibody or antigen binding fragment thereof that specifically binds IsdH comprises the amino acid sequence VH with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (including additions, deletions, and substitutions, such as conservative substitutions) in the amino acid sequence SEQ ID NO: 80, 82, 84, 86 or 88. In various embodiments, an antibody or antigen binding fragment thereof that specifically binds IsdH comprises the amino acid sequence VL with 1, 2, 3 , 4, 5, 6, 7, 8, 9 or 10 mutations (including additions, deletions and substitutions such as conservative substitutions) in the amino acid sequence of SEQ ID NO: 81, 83, 85, 87 or 89.

[0054] In certain embodiments, an antibody or antigen binding fragment that specifically binds to an IsdH surface antigen has one or more of the following characteristics:

(a) the dissociation constant (K _D ) for the S. aureus surface antigen is equal to about 100 nM or less, about 90 nM or less, about 80 nM or less, about 70 nM or less, about 60 nM or less, about 50 nM or less than about 40 nM or less, about 20 nM or less, about 10 nM or less, about 9, 8, 7, 6, 5, 4, 3, 2 or 1 nM. (or any value in between);

(b) reduces the ability of S. aureus to evade opsonophagocytosis by immune cells by at least 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therebetween), as measured by an opsonophagocytic assay killing;

(c) reduces the concentration of S. aureus colony-forming units (CFU) by at least 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therein) as measured by a bacteremia model ; or

(d) reduces immune cell infiltration, bacterial load, and release of proinflammatory cytokines.

[0055] The present antibodies and antigen-binding fragments thereof, which specifically bind surface antigens or secreted toxins of S. aureus, contain at least one antigen-binding domain that includes at least one complementarity determining region (for example, at least one of CDR1, CDR2 or CDR3). In some embodiments, the antibody or antigen binding fragment thereof comprises a VH that includes at least one CDR VH (eg, a CDR1 VH, a CDR2 VH, or a CDR3 VH). In certain embodiments, the antibody or antigen binding fragment thereof comprises a VL that includes at least one CDR VL (eg, CDR1 VL, CDR2 VL, or CDR3 VL). In some embodiments, the antibody or antigen binding fragment thereof comprises a VH that includes at least one VH CDR and at least one VL CDR.

[0056] The CDR regions disclosed herein can be combined in a variety of combinations because each CDR region can be independently selected and combined with another CDR region for a given antibody. In certain embodiments, the VH and/or VL CDR sequences may be present in any combination to form an antibody or antigen binding fragment thereof directed against a surface antigen or secreted toxin of S. aureus.

[0057] In some embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds IsdH comprises (a) a CDR1 VH comprising an amino acid sequence identical to or containing 1, 2 or 3 amino acid residue mutations relative to SEQ ID NO: 90 , 96, 102, 108 or 114; (b) CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 91, 97, 103, 109 or 115; and/or (c) a CDR3 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 92, 98, 104, 110 or 116.

[0058] In some embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds IsdH comprises (a) a CDR1 VL comprising an amino acid sequence identical to or containing 1, 2 or 3 amino acid residue mutations relative to SEQ ID NO: 93 , 99, 105, 111 or 117; (b) CDR2 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 94, 100, 106, 112 or 118; and/or (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 95, 101, 107, 113 or 119.

[0059] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds IsdH comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences identical to or containing 1, 2, or 3 mutations of amino acid residues in each CDR relative to: (a) CDR1 VH containing the amino acid sequence of SEQ ID NO: 90, 96, 102, 108 or 114; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 91, 97, 103, 109, or 115; (c) CDR3 VH containing the amino acid sequence of SEQ ID NO: 92, 98, 104, 110 or 116 (d) CDR1 VL containing the amino acid sequence of SEQ ID NO: 93, 99, 105, 111 or 117; (e) CDR2 VL containing the amino acid sequence of SEQ ID NO: 94, 100, 106, 112 or 118; and (f) CDR3 VL containing the amino acid sequence of SEQ ID NO: 95, 101, 107, 113 or 119.

[0060] In specific embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds IsdH comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL are selected from the group consisting of SEQ ID NO: 90, 91, 92, 93, 94 and 95; SEQ ID NO: 96, 97, 98, 99, 100 and 101; SEQ ID NO: 102, 103, 104, 105, 106 and 107; SEQ ID NO: 108, 109, 110, 111, 112 and 113; SEQ ID NOs: 114, 115, 116, 117, 118 and 119. In a further embodiment, the isolated antibody or antigen binding fragment thereof that specifically binds IsdH comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL, where CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL and correspond to SEQ ID NO: 90, 91, 92, 93, 94 and 95.

[0061] In some embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds to IsdH corresponds to any of the isolated antibodies or antigen binding fragments described above and has one or more of the following characteristics:

(a) dissociation constant (K _D ) for S. aureus surface antigen equal to about 100 nM or less, about 90 nM or less, about 80 nM or less, about 70 nM or less, about 60 nM or less, about 50 nM or less or about 40 nM or less, about 20 nM or less, about 10 nM or less, about 9, 8, 7, 6, 5, 4, 3, 2 or 1 nM (or any value in between);

(b) reduces the ability of S. aureus to evade opsonophagocytosis by immune cells by at least 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therebetween), as measured by an opsonophagocytic assay killing;

(c) reduces the number of S. aureus colony-forming units (CFU) by at least 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therein) as measured by a bacteremia model ; or

(d) reduces immune cell infiltration, bacterial load, and release of proinflammatory cytokines.

[0062] In additional embodiments, the present invention provides an isolated antibody or antigen binding fragment thereof that specifically binds to the same IsdH epitope as any other of the anti-ClfA antibodies or antigen binding fragments described above.

C. Antibodies to ClfA

[0063] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to the ClfA surface antigen comprises a heavy chain variable region (VH) that is 65%, 70%, 75%, 80%, 85%, 90% , 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 132 or 140. In certain embodiments, an antibody or antigen binding fragment thereof that specifically binds ClfA contains a light chain variable region (VL) that is 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 136 or 144. In specific embodiments, an antibody or antigen binding fragment thereof that specifically binds ClfA contain a VH that is 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 132 or 136 and a VL containing the amino acid sequence of SEQ ID NO : 140 or 144. In specific embodiments, an antibody or antigen binding fragment thereof that specifically binds to ClfA comprises VH and VL, wherein VH and VL are selected from the group consisting of SEQ ID NOs: 132 and 140; and SEQ ID NOs: 136 and 144. Example 7, Table 14 sets forth exemplary VH and VL sequences provided herein that may be present in any combination to form an antibody to a surface antigen or an antigen binding fragment thereof.

[0064] In additional embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds ClfA comprises a VH amino acid sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (including additions, deletions and substitutions, such as conservative substitutions) in the amino acid sequence of SEQ ID NO: 132 or 140. In various embodiments, an antibody or antigen binding fragment thereof that specifically binds ClfA comprises the amino acid sequence VL with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mutations (including additions, deletions and substitutions such as conservative substitutions) in the amino acid sequence of SEQ ID NO: 136 or 144.

[0065] In some embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds to ClfA comprises (a) a CDR1 VH comprising an amino acid sequence identical to or containing 1, 2 or 3 amino acid residue mutations relative to SEQ ID NO: 133 or 141; (b) CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 134 or 142; and/or (c) a CDR3 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 135 or 143.

[0066] In some embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds to ClfA comprises (a) a CDR1 VL comprising an amino acid sequence identical to or containing 1, 2 or 3 amino acid residue mutations relative to SEQ ID NO: 137 or 145; (b) CDR2 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 138 or 146; and/or (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid residue mutations relative to SEQ ID NO: 139 or 147.

[0067] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds ClfA comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences identical to or containing 1, 2, or 3 mutations of amino acid residues in each CDR with respect to: (a) CDR1 VH containing the amino acid sequence of SEQ ID NO: 133 or 141; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 134 or 142; (c) CDR3 VH containing the amino acid sequence of SEQ ID NO: 135 or 143; (d) CDR1 VL containing the amino acid sequence of SEQ ID NO: 137 or 145; (e) CDR2 VL containing the amino acid sequence of SEQ ID NO: 138 or 146; and (f) CDR3 VL containing the amino acid sequence of SEQ ID NO: 139 or 147.

[0068] In specific embodiments, an isolated antibody or antigen binding fragment thereof that specifically binds IsdH comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL are selected from the group consisting of SEQ ID NOs: 133, 134, 135, 137, 138 and 139 and SEQ ID NOs: 141, 142, 143, 144, 145, 146 and 147.

[0069] In additional embodiments, the present invention provides an isolated antibody or antigen binding fragment thereof that specifically binds to the same ClfA epitope as any other of the anti-ClfA antibodies or antigen binding fragments described above.

D. Antibodies to alpha toxin (AT)

[0070] In some embodiments, an antibody or antigen binding fragment thereof directed against a secreted toxin comprises a VH comprising the amino acid sequence SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62. In certain embodiments, the antibody or antigen binding fragment thereof to a secreted toxin comprises a VL comprising the amino acid sequence of SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63. In yet another embodiment, an anti-alpha toxin antibody or antigen binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62, and VL containing the amino acid sequence SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63. See Example 7, Table 7 for an illustration of the VH and VL sequences presented herein, which may be present in any combination to form an antibody or its alpha-toxin antigen-binding fragment or may be present in combination to form the mAb of the present invention. In some embodiments, VH is SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62. In various embodiments implementation VL is SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63.

[0071] Certain VH and VL nucleotide sequences encoding the VH and VL amino acid sequences discussed herein are presented in Example 7, Table 8.

[0072] In some embodiments, isolated antibodies or antigen binding fragments disclosed herein comprise VH and VL, wherein VH and VL have the amino acid sequences set forth in SEQ ID NOs: 20 and 19; SEQ ID NO: 22 and 21; SEQ ID NO: 24 and 23; SEQ ID NO: 26 and 25; SEQ ID NO: 28 and 27; SEQ ID NO: 41 and 42; SEQ ID NO: 43 and 44; SEQ ID NO: 45 and 46; SEQ ID NO: 47 and 48; SEQ ID NO: 47 and 48; SEQ ID NO: 49 and 50; SEQ ID NO: 51 and 52; SEQ ID NO: 51 and 52; SEQ ID NO: 53 and 54; SEQ ID NO: 55 and 56; SEQ ID NO: 57 and 58; SEQ ID NO: 59 and 60; SEQ ID NO: 61 and 58; SEQ ID NO: 62 and 58; SEQ ID NO: 62 and 63; SEQ ID NO: 79 and 63.

[0073] In certain embodiments, antibodies or fragments directed against surface antigens or secreted toxins of S. aureus contain VH and/or VL that have a predetermined percentage identity to at least one of the VH and/or VL sequences disclosed in the table 7.

[0074] In some embodiments, an antibody or antigen-binding fragment thereof to a secreted toxin comprises a VH amino acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or 100 % (or any percentage in between) identical to the amino acid sequence SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62. In certain embodiments, the antibody, or antigen binding fragment thereof, comprises a VH amino acid sequence with 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (including additions, deletions, and substitutions, such as conservative substitutions ) in the amino acid sequence SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62. As used herein, the expression “ "conservative substitution" refers to the substitution of a first amino acid with a second amino acid that substantially does not change the chemical, physical and/or functional properties of the antibody or antigen-binding fragment thereof (e.g., the antibody or antigen-binding fragment thereof retains the same charge, structure, polarity, hydrophobicity/hydrophilicity, and/or or retains functions such as the ability to bind alpha-toxin and thus reduce the virulence of S. aureus). In certain embodiments, the antibody or antigen binding fragment thereof comprises a VH amino acid sequence with a predetermined percentage identity to SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62 and have one or more of the following characteristics:

(a) a dissociation constant (K _D ) for S. aureus alpha toxin of approximately 13 nM or less;

(b) inhibit the formation of alpha-toxin oligomers by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween);

(c) reduce the cytolytic activity of the alpha toxin by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween) (for example, as determined by cell lysis assays and hemolysis); And

(d) reduce immune cell infiltration, bacterial load, and release of proinflammatory cytokines (eg, in an animal model of pneumonia).

[0075] In certain embodiments, an antibody or antigen-binding fragment thereof to a secreted toxin comprises a VL amino acid sequence that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% (or to any percentage in between) is identical to the amino acid sequence of SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63. In various embodiments an antibody or antigen binding fragment thereof, contains the amino acid sequence VL with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mutations (including additions, deletions and substitutions such as conservative substitutions) in the amino acid sequence SEQ ID NO : 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63. In certain embodiments, the antibody or antigen binding fragment thereof comprises a VL amino acid sequence with a predetermined percentage identity to SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63 and have one or more of the following characteristics:

(a) a dissociation constant (K _D ) for S. aureus alpha toxin of approximately 13 nM or less;

(b) inhibit the binding of alpha-toxin to the cell surface, thereby impairing the formation of alpha-toxin oligomers by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween) );

(c) reduce the cytolytic activity of the alpha toxin by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween) (for example, as determined by cell lysis assays and hemolysis); And

(d) reduce immune cell infiltration, bacterial load, and release of proinflammatory cytokines (eg, in an animal model of pneumonia).

[0076] In some embodiments, the isolated antibody or antigen-binding fragment specifically binds to a Staphylococcus aureus alpha-toxin polypeptide and includes (a) a CDR1 VH comprising an amino acid sequence identical to or containing 1, 2, or 3 amino acid residue mutations relative to SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 8, 11, 14, 17, 70 or 75; and/or (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72 , 76 or 78.

[0077] In specific embodiments, an isolated antibody or antigen-binding fragment that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, and CDR3 VH containing amino acid sequences identical to or containing 1, 2, or 3 amino acid residue mutations in each CDR with respect to SEQ ID NO: 7, 8 and 9; SEQ ID NO: 10, 11 and 12; SEQ ID NO: 13, 14 and 15; SEQ ID NO: 7, 17 and 18; SEQ ID NO: 7, 8 and 16; SEQ ID NO: 7, 8 and 65; SEQ ID NO: 7, 8 and 66; SEQ ID NO: 7, 8 and 67; SEQ ID NO: 7, 8 and 78; SEQ ID NO: 69, 70 and 71; SEQ ID NO: 7, 8 and 72; SEQ ID NO: 69, 75 and 71; SEQ ID NO: 69, 75 and 76 or SEQ ID NO: 69, 70 and 71.

[0078] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide includes (a) a CDR1 VL comprising an amino acid sequence identical to or containing 1, 2, or 3 amino acid residue mutations at relative to SEQ ID NO: 1 or 4; (b) CDR2 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 2, 5, 73 or 77; and/or (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 3, 6, 64, 68 or 74.

[0079] In specific embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences that are identical or contain 1, 2, or 3 amino acid residue mutations in each CDR with respect to SEQ ID NO: 1, 2 and 3; SEQ ID NO: 4, 5 and 6; SEQ ID NO: 1, 2 and 64; SEQ ID NO: 1, 2 and 68; SEQ ID NO: 1, 73 and 74 or SEQ ID NO: 1, 77 and 74.

[0080] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing identical amino acid sequences, or containing 1, 2 or 3 mutations of amino acid residues in each CDR, relative to: (a) CDR1 VH containing the amino acid sequence SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 8, 11, 14, 17, 70 or 75; (c) CDR3 VH containing the amino acid sequence SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76 or 78; (d) CDR1 VL containing the amino acid sequence of SEQ ID NO: 1 or 4; (e) CDR2 VL containing the amino acid sequence of SEQ ID NO: 2, 5, 73 or 77; or (f) a CDR3 VL comprising the amino acid sequence of SEQ ID NO: 3, 6, 64, 68 or 74.

[0081] In specific embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences identical to or containing 1, 2 or 3 mutations of amino acid residues in each CDR, relative to SEQ ID NO: 7, 8, 9, 1, 2 and 3; SEQ ID NO: 10, 11, 12, 1, 2 and 3; SEQ ID NO: 13, 14, 15, 4, 5 and 6; SEQ ID NO: 7, 17, 18, 1, 2 and 3; SEQ ID NO: 7, 8, 16, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 64; SEQ ID NO; 7, 8, 66, 1, 2 and 64; SEQ ID NO: 7, 8, 67, 1, 2 and 68; SEQ ID NO: 7, 8, 67, 1, 2 and 64; SEQ ID NO: 7, 8, 78, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 68; SEQ ID NO: 69, 70, 71, 1, 2 and 68; SEQ ID NO: 7, 8, 72, 1, 73 and 74; SEQ ID NO: 69, 75, 71, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 77 and 74; or SEQ ID NO: 69, 70, 71, 1, 77 and 74.

[0082] In some embodiments, a composition is provided comprising an isolated antibody or antigen binding fragment thereof that (i) includes a VH chain domain containing three CDRs and a VL chain domain containing three CDRs; and (ii) specifically bind to a Staphylococcus aureus alpha-toxin polypeptide, wherein the three CDR VH domains of the chain include (a) a VH CDR1 containing the amino acid sequence of SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 8, 11, 14, 17, 70 or 75; and (c) a CDR3 VH comprising the amino acid sequence of SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76, or 78. In specific embodiments, CDR1 VH, CDR2 VH, and CDR3 VH correspond to SEQ ID NO: 7, 8 and 9; SEQ ID NO: 10, 11 and 12; SEQ ID NO: 13, 14 and 15; SEQ ID NO: 7, 17 and 18; SEQ ID NO: 7, 8 and 16; SEQ ID NO: 7, 8 and 65; SEQ ID NO: 7, 8 and 66; SEQ ID NO: 7, 8 and 67; SEQ ID NO: 7, 8 and 78; SEQ ID NO: 69, 70 and 71; SEQ ID NO: 7, 8 and 72; SEQ ID NO: 69, 75 and 71; SEQ ID NO: 69, 75 and 76 or SEQ ID NO: 69, 70 and 71.

[0083] In certain embodiments, the antibody or antigen binding fragment thereof specifically binds to a secreted S. aureus toxin and comprises (a) a CDR1 VH comprising an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues relative to SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 8, 11, 14, 17, 70 or 75; and (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76 or 78; (d) CDR1 VL containing the amino acid sequence of SEQ ID NO: 1 or 4; (e) CDR2 VL containing the amino acid sequence of SEQ ID NO: 2, 5, 73 or 77; and (f) a CDR3 VL comprising the amino acid sequence of SEQ ID NO: 3, 6, 64, 68 or 74 and having one or more of the following characteristics:

(a) the dissociation constant (K _D ) for alpha toxin is approximately 13 nM or less;

(b) bind to alpha-toxin monomers;

(c) inhibit the formation of alpha-toxin oligomers by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween);

(d) reduce the cytolytic activity of the alpha toxin by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween) (for example, as determined by cell lysis assays and hemolysis); And

(f) reduce immune cell infiltration, bacterial load, and release of proinflammatory cytokines (eg, in an animal model of pneumonia).

[0084] In certain embodiments, the antibody or antibody fragment specifically binds to a surface antigen or secreted toxin of S. aureus and contains a heavy chain variable domain that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20, 22, 24, 26 , 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, 62, 80, 82, 84, 86, or 88, and contain a light chain variable domain that is at least 90% identical to the amino acid sequence of SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 63, 81, 83, 85, 87 or 89. In additional embodiments, the antibody or antigen binding fragment thereof reduces the ability of S. aureus to evade opsonophagocytosis by at least 50%. In additional embodiments, the antibody or antigen binding fragment thereof reduces the concentration of S. aureus CFU by at least 50%. In other embodiments, the antibody or antigen-binding fragment thereof inhibits the binding of one or more alpha-toxin monomers to each other (eg, inhibits oligomerization) and/or reduces the virulence of S. aureus.

[0085] In some embodiments, the isolated antibody or antigen-binding fragment specifically binds to a Staphylococcus aureus alpha-toxin polypeptide and includes (a) a CDR1 VH comprising an amino acid sequence identical to or containing 1, 2, or 3 amino acid residue mutations relative to SEQ ID NO : 7, 10, 13 or 69; (b) CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 8, 11, 14, 17, 70 or 75; and/or (c) CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72 , 76 or 78.

[0086] In specific embodiments, an isolated antibody or antigen-binding fragment that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, and CDR3 VH containing amino acid sequences identical to or containing 1, 2, or 3 amino acid residue mutations in each CDR with respect to SEQ ID NO: 7, 8 and 9; SEQ ID NO: 10, 11 and 12; SEQ ID NO: 13, 14 and 15; SEQ ID NO: 7, 17 and 18; SEQ ID NO: 7, 8 and 16; SEQ ID NO: 7, 8 and 65; SEQ ID NO: 7, 8 and 66; SEQ ID NO: 7, 8 and 67; SEQ ID NO: 7, 8 and 78; SEQ ID NO: 69, 70 and 71; SEQ ID NO: 7, 8 and 72; SEQ ID NO: 69, 75 and 71; SEQ ID NO: 69, 75 and 76 or SEQ ID NO: 69, 70 and 71.

[0087] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide comprises (a) a CDR1 VL comprising an amino acid sequence identical to or containing 1, 2, or 3 amino acid residue mutations relative to SEQ ID NO: 1 or 4; (b) CDR2 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 2, 5, 73 or 77; and/or (c) a CDR3 VL containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 3, 6, 64, 68 or 74.

[0088] In specific embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds to a Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences that are identical or contain 1, 2, or 3 amino acid residue mutations in each CDR with respect to SEQ ID NO: 1, 2 and 3; SEQ ID NO: 4, 5 and 6; SEQ ID NO: 1, 2 and 64; SEQ ID NO: 1, 2 and 68; SEQ ID NO: 1, 73 and 74 or SEQ ID NO: 1, 77 and 74.

[0089] In some embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing identical amino acid sequences, or containing 1, 2 or 3 mutations of amino acid residues in each CDR, relative to: (a) CDR1 VH containing the amino acid sequence SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 8, 11, 14, 17, 70 or 75; (c) a CDR3 VH containing the amino acid sequence of SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76 or 78; (d) CDR1 VL containing the amino acid sequence of SEQ ID NO: 1 or 4; (e) CDR2 VL containing the amino acid sequence of SEQ ID NO: 2, 5, 73 or 77; or (f) a CDR3 VL comprising the amino acid sequence of SEQ ID NO: 3, 6, 64, 68 or 74.

[0090] In specific embodiments, an isolated antibody or antigen-binding fragment thereof that specifically binds Staphylococcus aureus alpha-toxin polypeptide comprises CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL containing amino acid sequences identical to or containing 1, 2 or 3 mutations of amino acid residues in each CDR, relative to SEQ ID NO: 7, 8, 9, 1, 2 and 3; SEQ ID NO: 10, 11, 12, 1, 2 and 3; SEQ ID NO: 13, 14, 15, 4, 5 and 6; SEQ ID NO: 7, 17, 18, 1, 2 and 3; SEQ ID NO: 7, 8, 16, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 64; SEQ ID NO; 7, 8, 66, 1, 2 and 64; SEQ ID NO: 7, 8, 67, 1, 2 and 68; SEQ ID NO: 7, 8, 67, 1, 2 and 64; SEQ ID NO: 7, 8, 78, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 68; SEQ ID NO: 69, 70, 71, 1, 2 and 68; SEQ ID NO: 7, 8, 72, 1, 73 and 74; SEQ ID NO: 69, 75, 71, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 77 and 74; or SEQ ID NO: 69, 70, 71, 1, 77 and 74.

[0091] In some embodiments, a composition is provided comprising an isolated antibody or antigen binding fragment thereof that (i) comprises a VH chain domain containing three CDRs and a VL chain domain containing three CDRs; and (ii) specifically bind to a Staphylococcus aureus alpha-toxin polypeptide, wherein the three CDR VH domains of the chain include (a) a VH CDR1 containing the amino acid sequence of SEQ ID NO: 7, 10, 13 or 69; (b) CDR2 VH containing the amino acid sequence of SEQ ID NO: 8, 11, 14, 17, 70 or 75; and (c) a CDR3 VH comprising the amino acid sequence of SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76, or 78. In specific embodiments, CDR1 VH, CDR2 VH, and CDR3 VH correspond to SEQ ID NO: 7, 8 and 9; SEQ ID NO: 10, 11 and 12; SEQ ID NO: 13, 14 and 15; SEQ ID NO: 7, 17 and 18; SEQ ID NO: 7, 8 and 16; SEQ ID NO: 7, 8 and 65; SEQ ID NO: 7, 8 and 66; SEQ ID NO: 7, 8 and 67; SEQ ID NO: 7, 8 and 78; SEQ ID NO: 69, 70 and 71; SEQ ID NO: 7, 8 and 72; SEQ ID NO: 69, 75 and 71; SEQ ID NO: 69, 75 and 76 or SEQ ID NO: 69, 70 and 71.

[0092] In certain embodiments, the combination of CDR sequences present to form an antibody to a secreted toxin includes CDR1 VH containing SEQ ID NO: 7, 10, 13 or 69, CDR2 VH containing SEQ ID NO: 8, 11, 14, 17, 70, or 75, and a CDR3 VH containing SEQ ID NO: SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76, or 78, which are described in Table 9. B In some embodiments, CDR1 VL contains SEQ ID NO: 1 or 4, CDR2 VL contains SEQ ID NO: 2, 5, 73 or 77, and CDR3 VL contains SEQ ID NO: 3, 6, 64, 68 or 74, which are described in table 9.

[0093] The antibodies and antigen binding fragments disclosed herein may contain one or more amino acid sequences that are substantially similar to the amino acid sequences disclosed herein. Amino acid sequences that are substantially similar include sequences containing conservative amino acid substitutions as well as deletions and/or insertions of amino acids.

E. Wireframe areas

[0094] The variable domains of each of the heavy and light chains contain at least one of framework regions (FR1, FR2, FR3, FR4 or alternatively FW1, FW2, FW3, FW4). The heavy chain framework regions are designated FR VH, while the light chain framework regions are designated FR VL. In certain embodiments, the framework regions may include substitutions, insertions, or other changes. In certain embodiments, these changes result in improved or optimized binding affinity of the antibody. Non-limiting examples of framework region residues that may be modified include residues that non-covalently bind directly to the antigen, interact with/affect the conformation of the CDR, and/or participate in the formation of the VL-VH interface.

[0095] In certain embodiments, the framework region may contain one or more amino acid changes for “germline matching.” For example, the amino acid sequences of the heavy and light chains of a selected antibody can be compared to conform to the germline type, and in cases where certain framework residues of the selected VL and/or VH chains differ from the germline configuration (for example, as a result of somatic mutation of the immunoglobulin genes, used to produce a phage library), it may be desirable to “back-mutate” the modified framework residues of the selected antibodies to a germline configuration (i.e., alter the framework amino acid sequences of the selected antibodies so that they are similar to the germline framework amino acid sequences). Such "mutation to the original form" (or "germ-type conversion") of the framework residues can be accomplished using standard molecular biology techniques for introducing specific mutations (eg, site-directed mutagenesis; PCR-mediated mutagenesis). In some embodiments, the framework residues of the light and/or heavy chain variable regions are mutated back to the original species. In certain embodiments, the heavy chain variable region of an isolated antibody or antigen binding fragment disclosed herein is mutated back to the original species. In certain embodiments, the heavy chain variable region of the isolated antibody or antigen binding fragment contains at least one, at least two, at least three, at least four, or more mutations to the original species.

[0096] In certain embodiments, the VH antibodies to alpha toxin or an antigen binding fragment thereof may comprise FR1, FR2, FR3 and/or FR4 with amino acid sequences that are from about 65% to about 100% identical to the corresponding VH framework regions of SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62. In some embodiments, an anti-alpha toxin antibody or antigen-binding antibody thereof fragment contain an amino acid sequence of FR VH (FR1, FR2, FR3 and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage in between) corresponding FR regions VH SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62 In certain embodiments, the anti-alpha toxin antibody or antigen binding fragment thereof comprises an FR VH amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, 99% or 100% identical (or any percentage in between) to the corresponding FRs in VH SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62.

[0097] In certain embodiments, an anti-alpha toxin antibody or antigen binding fragment thereof may comprise a VH FR (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62. Specifically, each of FR1, FR2, FR3 or FR4 in VH may have an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations relative to the corresponding FR1, FR2, FR3 or FR4 in VH SEQ ID NO: 20, 22, 24, 26, 28, 41 , 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62.

[0098] In certain embodiments, the VL of an anti-alpha toxin antibody or antigen binding fragment thereof may comprise FR1, FR2, FR3 and/or FR4 with amino acid sequences that are from about 65% to about 100% identical to the corresponding framework regions within FR in VL SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63. In some embodiments, an anti-alpha toxin antibody or an antigen binding fragment thereof contain an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage in between) corresponding to the FR in VL SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63. B in certain embodiments, the anti-alpha toxin antibody or antigen binding fragment thereof comprises an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95 %, 96%, 97%, 98%, 99% or 100% identical (or any percentage in between) to the corresponding FRs in VL SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63.

[0099] In certain embodiments, an anti-alpha toxin antibody or antigen binding fragment thereof comprises a VL FR (FR1, FR2, FR3 and/or FR4) comprising an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations relative to the corresponding FR in VL SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63. Specifically, each of FR1, FR2, FR3 or FR4 in VL may have an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations with respect to the corresponding FR1, FR2, FR3 or FR4 in VH SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63.

[00100] In certain embodiments, an isolated antibody or antigen-binding fragment that specifically binds a secreted S. aureus toxin comprises a VH FR (FR1, FR2, FR3, and/or FR4) containing amino acid sequences identical to or containing 1, 2, or 3 amino acids mutations to the corresponding FR in VH SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62, and/ or FR VL (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations with respect to the corresponding FR in VL SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63.

[00101] In certain embodiments, the isolated antibody or antigen binding fragment thereof specifically binds a secreted S. aureus toxin and comprises a VH FR (FR1, FR2, FR3, and/or FR4) comprising an amino acid sequence identical to or containing 1, 2, or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62, and/or FR VL (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations with respect to the corresponding FR in VL SEQ ID NO: 19, 21, 23, 25, 27, 42 , 44, 46, 48, 50, 52, 54, 56, 58, 60, or 63, and wherein the antibody has one or more of the following characteristics:

(a) the affinity constant (K _D ) for alpha toxin is approximately 13 nM or less;

(b) binds to alpha-toxin monomers;

(c) inhibits the formation of alpha-toxin oligomers by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween);

(d) reduces the cytolytic activity of alpha toxin by at least 50%, 60%, 70%, 80%, 90% or 95% (or any percentage therebetween) (for example, as determined by cell lysis assays and hemolysis); And

(f) reduces immune cell infiltration, bacterial load, and release of proinflammatory cytokines (eg, in an animal model of pneumonia).

[00102] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds S. aureus IsdH surface antigen comprising the FR1, FR2, FR3 and/or FR4 VH regions having amino acid sequences that are from about 65% to about 100 % identical to the corresponding amino acid sequences of the four VH framework regions in SEQ ID NO: 80, 82, 84, 86 or 88. In some embodiments, the antibody or antigen binding fragment thereof comprises the amino acid sequence of a VH FR (FR1, FR2, FR3 and/or FR4), which is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage therein) to the corresponding amino acid sequences of the four FR regions VH SEQ ID NO: 80, 82, 84, 86, or 88. In certain embodiments, the antibody or antigen binding fragment thereof comprises an FR VH amino acid sequence (FR1, FR2, FR3, and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical (or any percentage in between) to the corresponding FR regions in VH SEQ ID NO: 80, 82, 84, 86 or 88.

[00103] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds IsdH comprising the FR1, FR2, FR3 and/or FR4 regions of the VL having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VL framework regions in SEQ ID NO: 81, 83, 85, 87, or 89. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL FR amino acid sequence (FR1, FR2, FR3, and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage in between) to the corresponding FR regions in VL SEQ ID NO: 81, 83, 85, 87 or 89. In certain embodiments, the antibody or antigen binding fragment thereof comprises an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95 %, 96%, 97%, 98%, 99% or 100% identical (or any percentage therein) to the corresponding FR regions in VL SEQ ID NO: 81, 83, 85, 87 or 89.

[00104] In certain embodiments, the isolated antibody or antigen binding fragment specifically binds IsdH and contains a VH FR (FR1, FR2, FR3, and/or FR4) containing an amino acid sequence identical to or containing 1, 2, or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 80, 82, 84, 86 or 88, and/or contain FR VL (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations relative to to the corresponding FR in VL SEQ ID NO: 81, 83, 85, 87 or 89.

[00105] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds IsdH comprising FR1, FR2, FR3 and/or FR4 VH regions having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VH framework regions in SEQ ID NO: 80, 82, 84, 86, or 88. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH FR amino acid sequence (FR1, FR2, FR3, and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage in between) to the corresponding amino acid sequences of the four VH FR regions SEQ ID NO: 80, 82, 84 , 86 or 88. In certain embodiments, the antibody or antigen binding fragment thereof comprises an FR VH amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, 99%, or 100% identical (or any percentage therein) to the corresponding FR regions in VH SEQ ID NO: 80, 82, 84, 86, or 88.

[00106] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds IsdH containing FR1, FR2, FR3 and/or FR4 VL regions having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VL framework regions in SEQ ID NO: 81, 83, 85, 87, or 89. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL FR amino acid sequence (FR1, FR2, FR3, and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage in between) to the corresponding FR regions in VL SEQ ID NO: 81, 83, 85, 87 or 89. In certain embodiments, the antibody or antigen binding fragment thereof comprises an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95 %, 96%, 97%, 98%, 99% or 100% identical (or any percentage therein) to the corresponding FR regions in VL SEQ ID NO: 81, 83, 85, 87 or 89.

[00107] In certain embodiments, the isolated antibody or antigen binding fragment specifically binds IsdH and contains a VH FR (FR1, FR2, FR3, and/or FR4) containing an amino acid sequence identical to or containing 1, 2, or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 80, 82, 84, 86 or 88, and/or contain FR VL (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations relative to to the corresponding FR in VL SEQ ID NO: 81, 83, 85, 87 or 89.

[00108] In certain embodiments, an isolated antibody or antigen-binding fragment is provided that specifically binds S. aureus ClfA surface antigen comprising the FR1, FR2, FR3 and/or FR4 VH regions having amino acid sequences that are from about 65% to about 100 % identical to the corresponding amino acid sequences of the four VH framework regions in SEQ ID NO: 132 or 140. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH FR amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 65 %, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage therein) to the corresponding amino acid sequences of the four FR regions of VH SEQ ID NO: 132 or 140. In certain in embodiments, the antibody or antigen binding fragment thereof comprises an FR VH (FR1, FR2, FR3 and/or FR4) amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical (or any percentage therein) to the corresponding FR regions in VH SEQ ID NO: 132 or 140.

[00109] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds ClfA comprising the FR1, FR2, FR3 and/or FR4 regions of the VL having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VL framework regions in SEQ ID NO: 136 or 144. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL FR amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to (or any percentage therebetween) the corresponding FR regions in VL SEQ ID NO: 136 or 144. In certain embodiments, the antibody or antigen binding fragment thereof contain an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% identical (or any percentage in between) to the corresponding FR regions in VL SEQ ID NO: 136 or 144.

[00110] In certain embodiments, the isolated antibody or antigen binding fragment specifically binds ClfA and contains a VH FR (FR1, FR2, FR3, and/or FR4) containing an amino acid sequence identical to or containing 1, 2, or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 132 or 136, and/or contain a VL FR (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations with respect to the corresponding FR in the VL SEQ ID NO: 136 or 144.

[00111] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds ClfA comprising FR1, FR2, FR3 and/or FR4 VH regions having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VH framework regions in SEQ ID NO: 132 or 140. In some embodiments, the antibody or antigen binding fragment thereof comprises a VH FR amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to (or any percentage therebetween) the corresponding amino acid sequences of the four FR regions of VH SEQ ID NO: 132 or 140. In certain embodiments, the antibody or the antigen binding fragment contains an amino acid sequence of FR VH (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% , 99% or 100% identical (or any percentage in between) to the corresponding FR regions in VH SEQ ID NO: 132 or 140.

[00112] In certain embodiments, an isolated antibody or antigen binding fragment is provided that specifically binds IsdH containing FR1, FR2, FR3 and/or FR4 VL regions having amino acid sequences that are from about 65% to about 100% identical to the corresponding amino acid sequences four VL framework regions in SEQ ID NO: 136 or 144. In some embodiments, the antibody or antigen binding fragment thereof comprises a VL FR amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to (or any percentage therebetween) the corresponding FR regions in VL SEQ ID NO: 136 or 144. In certain embodiments, the antibody or antigen binding fragment thereof contain an FR VL amino acid sequence (FR1, FR2, FR3 and/or FR4) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99 % or 100% identical (or any percentage in between) to the corresponding FR regions in VL SEQ ID NO: 136 or 144.

[00113] In certain embodiments, the isolated antibody or antigen binding fragment specifically binds ClfA and contains a VH FR (FR1, FR2, FR3, and/or FR4) containing an amino acid sequence identical to or containing 1, 2, or 3 amino acid mutations relative to the corresponding FR in VH SEQ ID NO: 132 or 140, and/or contain a VL FR (FR1, FR2, FR3 and/or FR4) containing an amino acid sequence identical to or containing 1, 2 or 3 amino acid mutations with respect to the corresponding FR in the VL SEQ ID NO: 136 or 144.

F. Nucleotide sequences encoding alpha-toxin antibodies and their antigen-binding fragments

[00114] In addition to the amino acid sequences described above, nucleotide sequences corresponding to the amino acid sequences disclosed herein are further provided. In some embodiments, the nucleotide sequence encodes an antibody or antigen-binding fragment thereof directed against a surface antigen or secreted toxin of S. aureus. Nucleotide sequences are shown in Example 7, Table 8. Thus, polynucleotide sequences encoding the VH and VL regions, including FR regions and CDRs, for the antibodies or fragments described herein, as well as expression vectors for their efficient expression in cells, are also provided. (for example, mammalian cells).

[00115] Also disclosed herein are polynucleotides that are substantially identical to polynucleotides encoding the amino acid sequences disclosed herein. Almost identical sequences can be polymorphic sequences, i.e. alternative sequences or alleles in a population. Substantially identical sequences may also contain mutagenized sequences, including sequences containing silent mutations. A mutation may involve changes to one or more residues, deletion of one or more residues, or insertion of one or more additional residues. Substantially identical sequences may also contain different nucleotide sequences that encode the same amino acid at any given amino acid position in the amino acid sequence disclosed herein due to the degeneracy of the nucleic acid code.

[00116] Also disclosed herein are polynucleotides that hybridize under high or low stringency hybridization conditions to polynucleotides that encode an antibody or antigen binding fragment thereof directed against a surface antigen or secreted toxin of S. aureus. As used herein, the term “stringency” refers to the experimental conditions (eg, temperature and salt concentration) of a hybridization experiment to determine the degree of homology between two nucleic acids; the higher the rigidity, the higher the percentage of homology between the two nucleic acids. As used herein, the phrase “hybridization” or grammatical variations thereof refers to the binding of a first nucleic acid molecule to a second nucleic acid molecule under low, medium or high stringency or nucleic acid synthesis conditions. Hybridization may include instances in which a first nucleic acid molecule binds to a second nucleic acid molecule, where the first and second nucleic acid molecules are complementary.

[00117] Stringent hybridization conditions include, but are not limited to, hybridization to filter-bound DNA in 6X sodium chloride/sodium citrate (SSC) at approximately 45 degrees Celsius followed by one or more washes in 0.2X SSC/0.1% SDS at approximately 50 -65 degrees Celsius. Other stringent conditions include hybridization to filter-bound DNA in 6X SSC at approximately 45 degrees Celsius followed by one or more washes in 0.1X SSC/0.2% SDS at approximately 65 degrees Celsius. Other stringent conditions are known to those skilled in the art and are included herein.

[00118] In certain embodiments, a nucleic acid disclosed herein may encode the amino acid sequence of an antibody or antigen-binding fragment thereof directed against a surface antigen or secreted toxin of S. aureus, or the nucleic acid may hybridize under stringent conditions with a nucleic acid comprising a nucleotide sequence which encodes the amino acid sequence of an antibody or an antigen-binding fragment thereof.

[00119] In certain embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence of an antibody or antigen binding fragment thereof that can bind a surface antigen or secreted toxin of S. aureus and that is at least about 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical to (or any percentage in between) the amino acid sequence of VH with SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, 62, 80, 82, 84, 86, or 88. In certain embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence with 1, 2, 3, 4 , 5, 6, 7, 8, 9 or 10 mutations (including additions, deletions and substitutions such as conservative substitutions) in the amino acid sequence SEQ ID NO: 20, 22, 24, 26, 28, 41, 43, 45, 47 , 49, 51, 53, 55, 57, 79, 59, 61, 62, 80, 82, 84, 86, or 88. In some embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence of an antibody or antigen-binding fragment thereof that can bind a surface antigen or secreted toxin of S. aureus and that are at least about 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage therein) VH nucleotide sequence with SEQ ID NO: 30, 32, 34, 36, 38, 120, 122, 124, 126 or 128.

[00120] In certain embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence of an antibody or antigen binding fragment thereof that can bind a surface antigen or secreted toxin of S. aureus and that is at least about 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical to (or any percentage in between) the amino acid sequence of VL with SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 63, 81, 83, 85, 87, or 89. In certain embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence with 1, 2, 3, 4, 5, 6 , 7, 8, 9 or 10 mutations (including additions, deletions and substitutions such as conservative substitutions) in the amino acid sequence SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52 , 54, 56, 58, 60, 63, 81, 83, 85, 87, or 89. In some embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence of an antibody or antigen-binding fragment thereof that can bind a surface antigen or secreted toxin S aureus and which are at least about 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% identical (or any percentage therebetween) to the nucleotide sequence of VL of SEQ ID NO: 29, 31, 33, 35, 37, 121, 123, 125, 127 or 129.

[00121] In particular embodiments, the polynucleotide sequence may comprise a nucleotide sequence encoding an amino acid sequence of an antibody or antigen binding fragment thereof that can bind a surface antigen or secreted toxin of S. aureus and that is at least about 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to (or any percentage in between) the amino acid sequence of VH and at least about 65%, 70%, 75%, 80%, 85%, 90% identical , 95% or 100% identical (or any percentage in between) to the amino acid sequence of VL, with the sequences of VH and VL presented under SEQ ID NO: 20 and 19; SEQ ID NO: 22 and 21; SEQ ID NO: 24 and 23; SEQ ID NO: 26 and 25; SEQ ID NO: 28 and 27; SEQ ID NO: 41 and 42; SEQ ID NO: 43 and 44; SEQ ID NO: 45 and 46; SEQ ID NO: 47 and 48; SEQ ID NO: 47 and 48; SEQ ID NO: 49 and 50; SEQ ID NO: 51 and 52; SEQ ID NO: 51 and 52; SEQ ID NO: 53 and 54; SEQ ID NO: 55 and 56; SEQ ID NO: 57 and 58; SEQ ID NO: 59 and 60; SEQ ID NO: 61 and 58; SEQ ID NO: 62 and 58; SEQ ID NO: 62 and 63; SEQ ID NO: 79 and 63; SEQ ID NO: 80 and 81; SEQ ID NO: 82 and 83; SEQ ID NO: 84 and 85; SEQ ID NO: 86 and 87; SEQ ID NO: 88 and 89.

[00122] The disclosed polynucleotides can be obtained and the nucleotide sequence of the polynucleotides can be determined using any method known in the art. For example, if the nucleotide sequence of an antibody is known, then the polynucleotide encoding the antibody can be assembled from chemically synthesized oligonucleotides. This will involve, for example, synthesizing overlapping oligonucleotides containing portions of the antibody coding sequence, annealing and ligating these oligonucleotides, and then amplifying the ligated oligonucleotides using PCR. The disclosed polynucleotides can also be obtained from any suitable nucleic acid source, such as an antibody cDNA library, or a cDNA library isolated from any tissue or cells expressing the antibody (eg, hybridoma cells selected to express the antibody).

G. Functional characteristics of antibodies or fragments directed against surface antigens or secreted toxins of S. aureus

[00123] In certain embodiments, an antibody or antigen-binding fragment thereof directed against a S. aureus surface antigen alters the biological properties of S. aureus cells that express the surface antigen. In various embodiments, the antibody binds a surface antigen of S. aureus, thereby enhancing opsonophagocytosis by host cells. In further embodiments, opsonophagocytosis is increased by 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therein) as measured by an opsonophagocytic killing assay. In some embodiments, binding of the antibody to the surface determinant antigen prevents the interaction of the surface antigen and the surface adhesin, thereby reducing the concentration of colony forming units (CFU) present in the host tissue, as measured in a mouse model of bacteremia. In further embodiments, the CFU concentration is reduced by 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage therein) compared to the CFU concentration in the presence of the negative control antibody or in the absence of the antibody or its antigen-binding fragment. For example, an anti-IsdH antibody may reduce the concentration of CFU by 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage in between). In some embodiments, an antibody to a surface antigen may compete with haptoglobin and/or hemoglobin for binding to S. aureus, thereby inhibiting the ability of S. aureus to access and utilize iron in hemoglobin. In certain embodiments, antibodies or fragments directed against a surface antigen reduce the ability of S. aureus to bind haptoglobin and/or hemoglobin by 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95 % (or any percentage in between) compared to S. aureus binding in the absence of antibody. For example, an anti-IsdH antibody may reduce the ability of S. aureus to bind haptoglobin and/or hemoglobin by 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% (or any percentage between them).

[00124] As used herein, the term “opsonophagocytic killing” (OPK) assay refers to any assay used to measure the percentage of phagocytic killing induced in host tissue in vitro following the addition of an antibody to a tissue sample containing S. aureus at a known concentration. This reduction in CFU is normalized against the control OPK level observed in the presence of the control antibody. The assay measures the ability of a target antibody to induce complement activation and subsequent phagocytosis. For example, the OPK may involve combining 10 μl of antibody and 10 μl of S. aureus (10 ⁶ cells/ml), followed by the addition of 10 μl of human promyelocytic leukemia (HL-60) cells (10 ⁷ cells/ml) and 10 μl of human serum, pre-absorbed by S. aureus. 10 µl of the mixture can then be inoculated (at time T ₀ ), followed by cell lysis with 1% saponin (at time T ₆₀ ) and determination of the S. aureus CFU concentration. The killing percentage can be calculated as follows: 100×(1-(T ₆₀ /T ₀ )), where T ₆₀ refers to the CFU concentration at the end of the assay (i.e., after 60 minutes) and T ₀ refers to the CFU concentration at the beginning of the analysis.

[00125] As used herein, the term “bacteremia model” refers to any in vivo model of S. aureus infection used to evaluate the effect of an antibody on the S. aureus bacterial load, expressed as a percentage reduction in CFU. For example, a bacteremia model may involve administration of a mouse antibody, subsequent intraperitoneal administration of 10 ⁸ CFU of S. aureus, and then drawing blood and measuring the concentration of CFU compared to the concentration of CFU after administration of a control antibody.

[00126] In certain embodiments, an anti-alpha toxin antibody or antigen-binding fragment thereof alters the biological properties of the alpha toxin and/or cells expressing the alpha toxin. In some embodiments, an anti-alpha toxin antibody or antigen-binding fragment thereof neutralizes the biological activity of an alpha-toxin by binding to a polypeptide and inhibiting membrane binding and assembly of alpha-toxin monomers into a transmembrane pore (eg, an alpha-toxin heptamer). Neutralization assays can be performed using methods known in the art, using, under certain conditions, commercially available reagents. Alpha toxin neutralization is often measured with an IC50 of 1x10 ^-6 M or less, 1x10 ^-7 M or less, 1x10 ^-8 M or less, 1x10 ^-9 M or less, 1x10 ^-10 M or less and 1×10 ^-11 M or less. The expression “50% inhibition concentration” (abbreviated as “IC50”) is the concentration of an inhibitor (e.g., an anti-alpha toxin antibody or antigen-binding fragment as provided herein) that is required to produce 50% inhibition of a given activity of a molecule targeted by the inhibitor ( for example, oligomerization of alpha-toxin to form a heptameric transmembrane pore complex). A lower IC50 value usually corresponds to a more potent inhibitor.

[00127] In certain embodiments, an anti-alpha toxin antibody or antigen-binding fragment thereof inhibits one or more biological activities of the alpha toxin. As used herein, the expression “inhibition” refers to a statistically significant reduction in biological activity, including complete blocking of activity. For example, “inhibition” may refer to a reduction in biological activity by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%, or any percentage therein. In certain embodiments, an anti-alpha toxin antibody or antigen binding fragment thereof inhibits one or more biological activities of the alpha toxin by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% or any percentage in between.

[00128] In some embodiments, an anti-alpha toxin antibody or antigen binding fragment thereof can eliminate alpha toxin secreted by pathogenic S. aureus. In some embodiments, the anti-alpha toxin antibody or antigen binding fragment thereof may reach at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100% elimination of alpha toxin secreted by S. aureus, or any percentage therebetween. In certain embodiments, substantially all of the detectable secreted alpha toxin is eliminated from cells infected with S. aureus.

[00129] In certain embodiments, an anti-alpha toxin antibody or an antigen-binding fragment thereof may inhibit the expression of one or more inducible genes that directly or indirectly respond to the environment created by S. aureus infection and/or alpha toxin expression and function. In specific embodiments, the anti-alpha toxin antibody or antigen-binding fragment thereof inhibits the expression of one or more inducible genes that respond directly or indirectly to the environment created by S. aureus alpha-toxin expression and function by at least 20% by at least 20%. by 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90% or any percentage between them.

H. Methods for producing antibodies to surface antigens and secreted toxins of S. aureus

[00130] The following describes exemplary methods for preparing the antibodies disclosed herein. In some embodiments, recombinant or hybridoma methods can be used to produce antibodies or fragments disclosed herein. In other embodiments, antibodies or antibody fragments can be isolated from phage antibody libraries generated using techniques known in the art. Other antibody production techniques known in the art can also be used to obtain antibodies to surface antigens or secreted toxins of S. aureus.

[00131] In some embodiments, anti-IsdH antibodies can be generated using native S. aureus IsdH, a mutant IsdH, a variant, or an antigenic fragment of IsdH. S. aureus cells expressing IsdH can also be used to produce antibodies. For use of IsdH in the production of antibodies to IsdH, it can also be produced recombinantly in isolated form from bacterial or eukaryotic cells using standard recombinant DNA techniques.

[00132] Polyclonal antibodies to a secreted toxin or surface antigen, such as IsdH, can be obtained using various procedures known in the art. For example, an IsdH polypeptide or an immunogenic fragment thereof can be administered to various animal hosts by subcutaneous or intraperitoneal injection of the appropriate antigen to induce the production of serum polyclonal antibodies specific for the antigen. Animal hosts include, but are not limited to, rabbits, mice and rats. In some embodiments, various adjuvants may be used to enhance the immune response depending on the host species and include, but are not limited to, Freund's adjuvant (complete and incomplete), mineral gels such as aluminum hydroxide, surfactants such as lysolecithin, polyols -pluronics, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, dinitrophenol and potentially useful human adjuvants such as BCG (Bacillus Calmette-Guerin) and Corynebacterium parvum. Other adjuvants known in the art can also be used.

[00133] Monoclonal antibodies to a secreted toxin or surface antigen, such as IsdH, can be produced using a wide variety of techniques known in the art, including the use of hybridoma, recombinant and phage display technologies, or combinations thereof. As used herein, the term “monoclonal antibody” refers to an antibody derived from a population of substantially homogeneous or isolated antibodies, eg, the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minute quantities. The modifier “monoclonal” should not be understood as requiring the antibody to be produced by any particular method. Monoclonal antibodies include mammalian monoclonal, chimeric, humanized, human, domain, diabodies, vaccibodies, linear and multispecific antibodies.

[00134] Once the antibody disclosed herein is produced, it can be purified by any method known in the art to purify the immunoglobulin molecule, such as by chromatography (e.g., ion exchange, affinity, and size exclusion column chromatography), centrifugation, differential solubility, or any other purification technique proteins. In addition, antibodies produced by the present technique, or fragments thereof, can be hybridized to heterologous polypeptide sequences (including epitope tags and other fusion proteins such as GST fusions) to facilitate purification and use of the antibody in downstream assays.

[00135] In certain embodiments, the antibodies disclosed herein are chimeric antibodies. Chimeric antibodies are antibodies in which part of the heavy and/or light chain is identical or homologous to the corresponding sequences of antibodies obtained from a particular species or belonging to a particular class or subclass of the antibody, while the other part of the chain(s) is identical or homologous to the corresponding sequences of antibodies obtained from antibodies of another type or belonging to another class or subclass, as well as fragments of such antibodies, provided that they exhibit the necessary biological activity. Chimeric antibodies disclosed herein include “primatized” antibodies containing variable domain antigen binding sequences derived from constant region sequences of a non-human primate (eg, a marmoset such as a baboon, rhesus monkey, or cynomolgus monkey) and a human. Chimeric antibodies disclosed herein also include humanized antibodies that are prepared using methods known in the art.

[00136] In other embodiments, the antibodies disclosed herein are human antibodies and are produced using methods known in the art. For example, fully human antibodies can be produced by introducing nucleic acids encoding functional human antibody loci into a rodent or other animal such that the rodent or other animal produces fully human antibodies. In another example, human antibodies can be produced using in vitro methods. Suitable examples include, but are not limited to, phage display, ribosomal display, yeast display, and other methods known in the art. Additional examples of methods for producing human antibodies or fragments directed against surface antigens or secreted toxins of S. aureus include the VelocImmune ^® mouse technique (Regeneron Pharmaceuticals). See, for example, US Pat. No. 6,596,541 (incorporated by reference in its entirety).

[00137] In certain embodiments, it may be necessary to reconvert the framework sequence of an antibody disclosed herein to a germline sequence, reconvert the CDR to a germline sequence, and/or remove the structural interference. Thus, in some embodiments, if a particular antibody disclosed herein differs from its corresponding germline sequence at the amino acid level, the antibody sequence may be backmutated to the germline sequence. Such corrective mutations can occur at one, two, three or more positions or a combination of any of the mutant positions using standard molecular biology techniques.

[00138] In certain embodiments, the present disclosure covers antibody fragments or antibodies containing these fragments. An antibody fragment contains a full-length portion of an antibody, which is typically its antigen-binding or variable region. Examples of such antibody fragments include Fab, Fab', F(ab')2, Fd and Fv fragments; diabodies; linear antibodies, single chain antibody molecules and multispecific antibodies are antibodies derived from these fragments.

[00139] In addition to the human, humanized and/or chimeric antibodies described above, the antibodies disclosed herein can also be further modified to include one or more of the following: substitution, addition and/or deletion of at least one amino acid residue and/or polypeptide in the VL -domain, and/or VH domain, and/or Fc region and post-translational modifications. Any combination of deletions, insertions and substitutions can be performed to produce the final construct, provided that the final construct has the required characteristics.

[00140] Such modifications include antibody conjugates, wherein the antibody is covalently attached to a moiety. Components suitable for attachment to antibodies include, but are not limited to, proteins, peptides, drugs, tags and cytotoxins. Such antibody modifications can be made to modify or optimize the characteristics (eg, biochemical, binding and/or functional) of antibodies that are useful for detecting, diagnosing and/or treating S. aureus infection and related diseases or disorders. Methods for preparing conjugates, performing amino acid and/or polypeptide changes and post-translational modifications are known in the art. Also included in these modifications are hybrid proteins, i.e., an antibody or fragment thereof hybridized with a heterologous protein, polypeptide or peptide.

[00141] In certain embodiments, antibodies or fragments directed against surface antigens or secreted toxins of S. aureus are prepared to include an altered Fc region (also referred to herein as a “variant Fc region”) in which the Fc region has been one or more changes to change the functional and/or pharmacokinetic properties of antibodies. Such changes may result in altered effector function, reduced immunogenicity and/or increased serum half-life. In certain embodiments, the effector function of an antibody can be modified through changes in the Fc region, including, without limitation, amino acid substitutions, amino acid additions, amino acid deletions, and changes in post-translational modifications of amino acids in the Fc (eg, glycosylation).

[00142] In some embodiments, an Fc variant antibody is produced that has altered binding properties to an Fc ligand (eg, an Fc receptor such as C1q) compared to a native Fc antibody. Non-limiting examples of binding properties include binding specificity, equilibrium dissociation constant ( _Kd ), dissociation and association rates (koff _{and kon} , respectively), binding affinity and/or avidity.

[00143] In certain embodiments, antibodies disclosed herein are glycosylated to alter the effector function of the antibodies or alter the affinity of the antibody for a target antigen. In some embodiments, the glycosylation pattern in the variable region of the antibody is modified. For example, a non-glycosylated antibody (ie, an antibody that has not been glycosylated) can be prepared. Glycosylation can be modified, for example, to increase the affinity of the antibody for a target antigen. Such carbohydrate modifications can be achieved by, for example, changing one or more glycosylation sites within the antibody sequence. For example, one or more amino acid mutations can be made that result in the removal of one or more glycosylation sites of the framework region of the variable region, thereby eliminating glycosylation at that site.

[00144] In certain embodiments, antibodies disclosed herein are conjugated or covalently attached to another substance using methods known in the art. In some embodiments, the attached substance is a detectable label (also referred to herein as a reporter molecule) or a solid support. Suitable substances for attachment to an antibody include, but are not limited to, amino acid, peptide, protein, polysaccharide, nucleoside, nucleotide, oligonucleotide, nucleic acid, hapten, drug, hormone, lipid, lipid array, synthetic polymer, polymer microparticle, biological cell, virus, fluorophore , chromophore, dye, toxin, hapten, enzyme, antibody, antibody fragment, radioactive isotope, solid matrices, semi-solid matrices and combinations thereof. Methods for conjugating or covalently attaching another substance to an antibody are known in the art.

I. Methods of treatment using a surface antibody to an antigen or secreted toxin of S. aureus or fragments thereof

[00145] Antibodies or fragments disclosed herein can be administered alone, in combination with each other, or in combination with additional pharmaceutical agents, such as antibiotics, to prevent S. aureus infections or associated symptoms or conditions (for example, to treat hyperinflammation induced by alpha toxin). Antibodies and combinations of antibodies or fragments can be used to treat or prevent a wide range of conditions/diseases, including both chronic and acute conditions such as, but not limited to, bacteremia, burns, cellulitis, dermatotonecrosis, eyelid infections, food poisoning, joint infections, neonatal conjunctivitis , osteomyelitis, pneumonia, skin infections, post-operative wound infection, scalded skin syndrome, endocarditis, meningitis, abscess formation and toxic shock syndrome. Below are additional details regarding possible diseases/conditions eligible for anti-S. aureus therapy.

[00146] In certain embodiments, at least one antibody disclosed herein can be administered in combination with at least one additional therapeutic agent (eg, an antibiotic). Examples of antibiotics that can be administered in combination include penicillin, oxacillin, flucloxacillin, vancomycin and gentamicin. In certain embodiments, combination therapy with an antibiotic and at least one antibody or antigen-binding fragment disclosed herein increases the effectiveness of treatment by, for example, reducing the concentration of S. aureus CFU in host tissue, reducing the ability of S. aureus to evade opsonophagocytosis, and/ or reduction in S. aureus virulence compared with antibody therapy alone.

[00147] Combination therapy (eg, treatment or prevention with multiple antibodies) may provide an advantage over individual therapy by addressing multiple non-overlapping therapeutic targets of S. aureus. For example, an antibody targeting a secreted toxin can neutralize the deleterious effects of the toxin, such as alpha-toxin-induced hyperinflammation. Conversely, a coadministered antibody targeting a surface antigen (eg, IsdH) can inhibit colony growth and evasion of opsonophagocytosis in S. aureus, which are not altered by an antibody targeting a secreted toxin. Combination therapy can also ensure that the therapy is effective against a wider range of S. aureus strains or mutants, some of which may lack the antigenic target of a particular antibody.

[00148] In particular, the combination therapy may include one or more antibodies or antigen-binding fragments thereof that specifically bind to a surface determinant, such as SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH or PNAG, and one or more antibodies or antigen-binding fragments thereof that bind to a secreted toxin, such as alpha toxin (AT). In specific embodiments, the combination therapy may include an antibody or antigen-binding fragment thereof that specifically binds IsdH and an antibody or antigen-binding fragment thereof that specifically binds AT; an antibody or an antigen-binding fragment thereof that specifically binds to ClfA, and an antibody or an antigen-binding fragment thereof that specifically binds to AT; an antibody or an antigen-binding fragment thereof that specifically binds to IsdH, and an antibody or an antigen-binding fragment thereof that specifically binds to AT; an antibody or an antigen-binding fragment thereof that specifically binds to ClfA, and an antibody or an antigen-binding fragment thereof that specifically binds to IsdH; or an antibody or antigen-binding fragment thereof that specifically binds to IsdH, an antibody or antigen-binding fragment thereof that specifically binds to ClfA, and an antibody or antigen-binding fragment thereof that specifically binds to AT. In specific embodiments, the combination therapy may include an antibody or antigen-binding fragment thereof that specifically binds IsdH and an antibody or antigen-binding fragment thereof that specifically binds AT, wherein the anti-IsdH antibody or fragment thereof comprises VH and/or VL or CDR1 VH , CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL mAb 2F4, and the anti-AT antibody or fragment thereof contains VH and/or VL or CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL mAb LC10 or contain SEQ ID NO: 130 and SEQ ID NO: 131.

[00149] In specific embodiments, anti-AT antibodies or antigen-binding fragments thereof may comprise VH and/or VL or CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL, and CDR3 VL of any of the antibodies listed in Table 7 or 10, antibodies anti-IsdH or antigen-binding fragments thereof may contain VH and/or VL or CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL of any of the antibodies listed in Table 12, and anti-ClfA antibodies or antigen-binding fragments thereof may contain VH and/or VL or CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL of any of the antibodies listed in Table 14.

[00150] In various embodiments, the disclosed antibodies, combinations of antibodies, and/or combinations of antibodies and antibiotics can be administered therapeutically to treat S. aureus infection, or prophylactically to prevent infection. For example, combination therapy may be given before surgery to prevent a complication caused by S. aureus, or after surgery to treat an S. aureus infection that was acquired during surgery.

[00151] Also provided herein are pharmaceutical compositions for use in the treatment of S. aureus infections or as prophylaxis. In several embodiments, the pharmaceutical composition contains at least one antibody disclosed herein and a pharmaceutically acceptable carrier. The expression “pharmaceutically acceptable carrier” means one or more non-toxic materials that do not interfere with the effectiveness or biological activity of the active ingredients. Such preparations may contain salts, buffers, preservatives, compatible carriers, and optionally other therapeutic agents. Such pharmaceutically acceptable preparations may also contain compatible solid or liquid excipients, diluents or encapsulating agents that are suitable for administration to the human body. The expression "carrier" means an organic or inorganic ingredient, natural or synthetic, with which the active ingredient is combined to facilitate pharmaceutical use.

[00152] Therapeutic compositions according to the present method can be formulated at a specific dosage. Dosing regimens may be adjusted to provide the optimal desired response (eg, therapeutic response). For example, a single bolus may be administered, multiple divided doses may be administered over a period, or the dose may be proportionally decreased or increased as determined by the needs of the therapeutic situation. In some embodiments, the selected dosage is suitable for intravenous, intramuscular, intranasal, oral, topical, or subcutaneous delivery. The amount of active ingredient that can be combined with the carrier material to form a unit dosage form will vary depending on the subject being treated and the particular route of administration.

[00153] Also disclosed herein is a pharmaceutical kit for therapeutic use in the treatment of S. aureus infection or as prophylaxis against such infection. In some embodiments, the kit includes one or more containers filled with a sterile liquid therapeutic composition or lyophilized composition containing at least one antibody or antigen binding fragment disclosed herein and a pharmaceutically acceptable carrier. In some embodiments, the container filled with the liquid composition is a pre-filled syringe. In other embodiments, a container filled with a sterile lyophilized powder formulation is suitable for reconstitution and subsequent administration. In certain embodiments, the formulations comprise antibodies and antigen-binding fragments thereof recombinantly hybridized or chemically conjugated to at least one component, including, but not limited to, a heterologous protein, heterologous polypeptide, heterologous peptide, large molecule, small molecule, marker sequence, diagnostic or detectable agent, therapeutic component, drug component, radioactive metal ion, second antibody and solid support. In certain embodiments, the formulations are formulated in single-dose vials as sterile liquids. In some embodiments, the composition is provided in the form of a pre-filled syringe.

J. Diseases associated with S. aureus infection

[00154] The antibodies and antigen binding fragments disclosed herein can be used for the detection, diagnosis, prevention and/or treatment of disease associated with S. aureus infection. Antibodies can also be used to reduce and/or prevent one or more symptoms associated with S. aureus infection.

[00155] Also provided herein is a method of preventing, treating, or controlling pneumonia in a subject, comprising administering a composition that includes an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof, to a person in need thereof. subject in an amount effective to prevent, treat or control pneumonia.

[00156] As used herein, the expressions “treat,” “treat,” or “treatment” may refer to therapeutic treatments and prophylactic or preventative measures where the goal is to prevent or slow (attenuate) an undesirable physiological change or disorder, such as progression diseases. Beneficial or desirable clinical outcomes include, but are not limited to, relief of symptoms, reduction in disease severity, stabilization (ie, non-worsening) of disease state, delay or slowing of disease progression, improvement or alleviation of disease state. “Treatment” may also mean an increase in survival compared to the expected survival if treatment had not been received. Those who need treatment include those who already have a condition or disorder, those who are likely to have a condition or disorder, or those who need to be prevented from developing a condition or disorder.

[00157] Also provided in some embodiments is a method of preventing, treating, or controlling a condition caused by a skin infection in a subject, comprising administering a composition that includes an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or the combination thereof of the present invention required by the subject in an amount effective to prevent, treat or control a condition caused by a skin infection. In certain embodiments, the condition caused by the skin infection is dermatonecrosis. In some embodiments, the condition caused by a skin infection includes infection of the skin with S. aureus. In certain embodiments, the method prevents a condition caused by a skin infection.

[00158] In some embodiments, a method is provided for preventing, treating, or controlling S. aureus infection associated with dialysis treatment, high-risk surgery, pneumonia, ventilator-associated pneumonia (VAP), or reinfection following prior hospital discharge from a prior treatment. or surgery, which involves administering to a subject in need thereof a composition that includes an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or combinations thereof.

[00159] Some embodiments also provide a method of preventing, treating, or controlling a condition associated with S. aureus infection, which includes administering a composition that isolated includes an antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, to the subject in need thereof in an amount effective to reduce cell lysis. In certain embodiments, the method prevents a condition associated with S. aureus infection. In some embodiments, the cell is a red blood cell from the blood or lung.

[00160] Provided herein are methods of preventing or reducing the severity of sepsis associated with S. aureus in a mammalian subject, comprising administering to the subject an effective amount of an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or their combinations. Also provided are methods of reducing the bacterial load of S. aureus in the bloodstream or heart of a mammalian subject, comprising administering to the subject an effective amount of an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or a combination thereof. Also provided are methods for reducing agglutination of S. aureus bacteria and/or formation of thromboembolic lesions in a mammalian subject, comprising administering to the subject an effective amount of an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or a combination thereof.

[00161] Methods of preventing S. aureus-associated sepsis in a mammalian subject preferably involve administering to the subject, prior to the infection event, an effective amount of an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof. As used herein, the expression “infection event” refers to an episode in which a subject is, or may have been, exposed to S. aureus infection. Exemplary infection events include, but are not limited to, surgery on any part of the body, including the mouth, hands, arms, legs, torso, internal organs (eg, heart, brain, intestines, kidneys, stomach, lungs, liver, spleen, pancreas, and etc.), bones, skin. Surgery exposes conditions, such as open surgical wounds and organs, that can be easily infected by S. aureus. Additional infection events include trauma to any part of the body that creates open wounds or other access to the bloodstream through which S. aureus infection can enter the body. Additional infection events include blood transfusions, injections of drugs or illegal or legal drugs, needlesticks, tattoo needles, insertion or presence of intravenous (IV) catheters, insertion and presence of surgical drains, and the presence of areas of skin breakdown such as pressure ulcers (decubital ulcers). ulcers).

[00162] In embodiments in which the methods provide prevention of S. aureus-associated sepsis, an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof, is preferably administered at least 1 hour before the infection event. For example, at least 1 hour before surgery (infection event). Preferably, an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a toxin or surface determinant of S. aureus, or a combination thereof, is administered at least 6 hours before, at least 12 hours before, at least 18 hours before, at least 24 hours, at least 30 hours, at least 36 hours, at least 42 hours, at least 48 hours or earlier before the infection event. In embodiments, an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof, is preferably administered over about 6 hours to about 36 hours, about 6 hours to about 36 hours, about 12 hours to about 36 hours, from about 12 hours to about 24 hours, from about 24 hours to about 36 hours, from about 20 hours to about 30 hours, from about 20 hours to about 28 hours, from about 22 hours to about 26 hours or approximately 12 hours, approximately 13 hours, approximately 14 hours, approximately 15 hours, approximately 16 hours, approximately 17 hours, approximately 18 hours, approximately 19 hours, approximately 20 hours, approximately 21 hours , approximately 22 hours, approximately 23 hours, approximately 24 hours, approximately 25 hours, approximately 26 hours, approximately 27 hours, approximately 28 hours, approximately 29 hours, or approximately 30 hours, or approximately 31 hours, or approximately 32 hours, or approximately 33 hours, or approximately 34 hours, or approximately 35 hours, or approximately 36 hours before the infection event.

[00163] As used herein, “prevention” of S. aureus-related sepsis refers to reducing the risk of a subject experiencing S. aureus-related sepsis during an infection event. Preferably, the subject's risk of S. aureus-related sepsis is reduced by at least 30% compared to a subject who has not been administered, prior to the infection event, an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant , or a combination thereof. More preferably, the risk is reduced by at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or the risk is completely eliminated compared to a subject who, prior to the infection event, has not been administered an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof.

[00164] In methods of reducing the severity of S. aureus-associated sepsis in a mammalian subject, such methods preferably comprise administering an effective amount of an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant, or a combination thereof, a subject who exhibits symptoms of sepsis associated with S. aureus. Such symptoms may include, for example, chills, confusion or delirium, fever or low body temperature (hypothermia), dizziness due to low blood pressure, rapid heartbeat, tremors, skin rash and hot skin.

[00165] As used herein, the expression “reduction in severity” when applied to sepsis refers to a reduction in the intensity of symptoms observed in a subject who has experienced sepsis associated with S. aureus. Preferably, the intensity of symptoms is reduced by at least 30% compared to the intensity of symptoms observed in a subject who also has S. aureus-associated sepsis, but the subject has not been administered an isolated antibody or antigen-binding fragment thereof that is immunospecific bind to a toxin or surface determinant of S. aureus or a combination thereof. More preferably, the intensity of symptoms is reduced by at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or symptoms completely are eliminated (ie, the subject is cured of the infection and sepsis) compared to a subject who, prior to the infection event, has not been administered an isolated antibody or antigen-binding fragment thereof that immunospecifically binds to a S. aureus toxin or surface determinant or a combination thereof.

[00166] Non-limiting examples of some commonly known conditions caused by S. aureus infection include burns, cellulitis, dermatotonecrosis, eyelid infections, food poisoning, joint infections, pneumonia, skin infections, surgical wound infections, scalded skin syndrome, and toxic shock syndrome. In addition, it is a common pathogen in infections associated with foreign bodies such as intravascular catheters, pacemakers, artificial heart valves, and prosthetic joints. Some of the conditions or diseases caused by S. aureus are described below. Some or all of the conditions and diseases described below may involve the direct effect of secreted toxins as part of an infection or mediator of a condition or disease state, while some or all of the conditions may involve indirect or concomitant effects of secreted toxins (eg, as underlying factors virulence agents that cause the main symptom or majority of the symptoms associated with the condition, or as agents that act as further exacerbators of the disease due to disruption of cellular function or cell lysis).

a) Burns

[00167] Burn wounds are often initially sterile. However, moderate to severe burns typically damage physical and immune barriers to infection (eg, blistering, cracking, or peeling of the skin), causing loss of fluid and electrolytes, and resulting in local or general physiological dysfunction. Contact of damaged skin with viable bacteria may result in mixed colonization at the site of injury. Infection may be limited to nonviable debris on the surface of the burn (“burn eschar”), or colonization may develop into a deep cutaneous infection and involve viable tissue below the burn eschar. More severe infections may penetrate under the skin, enter the lymphatic system and/or bloodstream and develop into septicemia. S. aureus is typically found among the pathogens that colonize burn wound infections. S. aureus can destroy granulation tissue and cause severe septicemia.

b) Phlegmon

[00168] Cellulitis is an acute skin infection that often begins as a superficial infection that can spread beneath the dermal layer. Cellulitis is most often caused by a mixed infection of S. aureus in combination with S. pyogenes. Cellulitis can lead to systemic infection.

c) Dermatonecrosis

[00169] Dermatonecrosis is an infection of the skin and subcutaneous tissues that readily spreads along the fascial plane within the subcutaneous tissue. This condition causes the upper and/or lower layers of the skin to become necrotic, and it can spread to the underlying and surrounding tissues.

d) Necrotizing fasciitis

[00170] Necrotizing fasciitis is called “flesh-eating disease” or “flesh-eating bacteria syndrome.” Necrotizing fasciitis can be caused by a polymicrobial infection (eg, type I, caused by a mixed bacterial infection) or a monomicrobial infection (eg, type II, caused by a single pathogenic strain of bacteria). Many types of bacteria can cause necrotizing fasciitis, non-limiting examples include group A streptococcus (eg, Streptococcus pyrogenes), Staphylococcus aureus, Vibrio vulnificus, Clostridium perfringens, and Bacteroides fragilis. Those most likely to suffer from dermatotonecrosis (eg, necrotizing fasciitis) are those with a suppressed or damaged immune system.

[00171] Initially, group A streptococcus was diagnosed as the cause of most cases of type II dermatonecrotizing infections. However, since 2001, monomicrobial necrotizing fasciitis has been observed to be caused by methicillin-resistant Staphylococcus aureus (MRSA) with increasing frequency. The infection begins locally, sometimes at the site of an injury, which may be severe (such as the result of surgery), minor, or even invisible. Patients usually complain of severe pain, which may seem excessive given the appearance of the skin. As the disease progresses, the tissue becomes swollen, often within a few hours. Diarrhea and vomiting are also common symptoms.

[00172] Signs of inflammation may not be noticeable in the early stages of infection if the bacteria are deep in the tissue. If the bacteria are shallow, signs of inflammation such as redness and swelling or hot skin will quickly appear. The skin color may turn purple and blisters may form, followed by necrosis (eg, death) of the subcutaneous tissue. Patients with necrotizing fasciitis typically have a fever or appear very ill. It has been noted that if left untreated, mortality rates are as high as 73 percent.

e) Pneumonia

[00173] S. aureus has also been identified as a cause of staphylococcal pneumonia. Staph pneumonia causes inflammation and swelling of the lung, which in turn causes fluid to accumulate in the lung. Fluid accumulation in the lung can prevent oxygen from entering the bloodstream. People with influenza are at risk of developing bacterial pneumonia. Staphylococcus aureus is the most common cause of bacterial pneumonia in patients who already have the flu. Typical symptoms of staph pneumonia include cough, difficulty breathing and fever. Additional symptoms include fatigue, yellow or bloody mucus, and chest pain that gets worse with breathing. Methicillin-resistant S. aureus (MRSA) is increasingly being diagnosed as the strain identified in staphylococcal pneumonia.

f) Infections of postoperative wounds

[00174] Postoperative wounds often penetrate deep into the body. Therefore, when a wound becomes infected, infection of such wounds poses a serious risk to the patient. Often the causative agent of infections in postoperative wounds is S. aureus. S. aureus is unusually capable of penetrating surgical wounds; suture wounds may become infected with far fewer S. aureus cells than are necessary to cause infection in normal skin. Penetration of surgical wounds can lead to severe septicemia caused by S. aureus. Penetration of S. aureus into the bloodstream can lead to dissemination and infection of internal organs, particularly heart valves and bone, causing systemic diseases such as endocarditis and osteomyelitis.

g) Scalded skin syndrome

[00175] S. aureus may be a major causative agent of “scalded skin syndrome,” also called “staphylococcal scalded skin syndrome,” “toxic epidermal necrosis,” “localized bullous impetigo,” “Ritter's disease,” and “Lyell's disease.” Scalded skin syndrome occurs frequently in adolescents, typically in outbreaks caused by the development of strains of S. aureus that produce epidermolytic exotoxins (eg, exfoliatin A and B, sometimes called scalded skin syndrome toxin) that cause delamination of the epidermal layer. One of the exotoxins is encoded by the bacterial chromosome, and the other is encoded by a plasmid. Exotoxins are proteases that break down desmoglein-1, which normally holds the granulosa and spinous layers of the skin together.

[00176] The bacteria may initially infect only a minor affected area, but the toxin disrupts intercellular junctions, spreads throughout the epidermal layers and allows the infection to penetrate the outer layer of the skin, leading to the desquamation that characterizes the disease. Peeling of the outer layer of skin usually exposes the underlying normal skin, but if left untreated, the loss of fluid in the process can cause severe damage in young children.

h) Toxic shock syndrome

[00177] Toxic shock syndrome (TSS) is caused by strains of S. aureus that produce the so-called “toxic shock syndrome toxin.” The disease can be caused by S. aureus infection anywhere, but is often mistakenly thought to be limited to women who use tampons. The disease includes toxemia and septicemia and can be fatal.

[00178] Symptoms of toxic shock syndrome vary depending on the underlying cause. TSS, which results from infection with the bacteria Staphylococcus aureus, typically presents in otherwise healthy individuals with high fever accompanied by low blood pressure, discomfort, and confusion, which can quickly progress to confusion, coma, and multi-organ failure. The characteristic rash, often seen early in the disease, resembles a sunburn and can affect any area of the body, including the lips, mouth, eyes, palms and soles of the feet. In patients who survive the initial bout of infection, the rash will desquamate or peel off within 10 to 14 days.

[00179] As stated above, due to the increasing number of multidrug-resistant S. aureus strains, an increasing number of antibiotics that have been widely used to treat S. aureus infections no longer control or eliminate methicillin-resistant and multidrug-resistant Staphylococcus infections aureus. Antibodies to surface determinants and secreted toxins of S. aureus described herein may help reduce the severity of infection and may also help eliminate, prevent (prophylactically), or reduce the amount of pathogenic S. aureus in an infected host. The antibodies can also be used to detect S. aureus and, in the case of a patient sample, diagnose S. aureus infections.

K. Methods for detecting S. aureus using antibodies or fragments directed against surface antigens or secreted toxins of S. aureus

[00180] In various embodiments, antibodies disclosed herein can be used alone or in combination to detect the presence of S. aureus in a sample.

[00181] In certain embodiments, the method includes contacting a test sample with one of the isolated antibodies or fragments disclosed herein. The antibody or antigen-binding fragment then binds to a surface antigen or secreted toxin from S. aureus to form an antigen-antibody complex. In additional embodiments, the method includes contacting an antigen-antibody complex with a detectable label, wherein the signal produced by the detectable label is directly correlated with the presence of S. aureus in the sample. For example, the detectable label may include one or more fluorescent markers that bind an antibody or antigen in an antibody-antigen complex such that an increase in fluorescence correlates with an increased concentration of S. aureus or secreted toxin in the sample.

[00182] In other embodiments, the detectable label competes with a S. aureus surface antigen or secreted toxin for binding to the antibody or antigen-binding fragment thereof, wherein the signal produced by the detectable label indirectly correlates with the concentration of S. aureus or secreted toxin in the sample. For example, the detectable label may include one or more fluorescent markers that compete with a surface antigen or secreted toxin for binding to the antibody such that a decrease in fluorescence correlates with an increased concentration of S. aureus or secreted toxin in the sample.

[00183] In certain embodiments, the detectable signal produced by the detectable label in the test sample is compared with the signal from at least one control sample with a known concentration of antigen and antibody. In embodiments using control samples, the antibody-antigen complex is detected in the control and test samples using a detectable label, and any statistically significant difference in the detected signal among samples indicates the concentration, presence or absence of S. aureus and/or secreted toxin in the test sample.

[00184] In other embodiments, a combination of antibodies is used to detect S. aureus in a sample. In various embodiments, the method includes contacting a test sample with an isolated antibody or antigen-binding fragment thereof directed against a S. aureus surface antigen and an isolated antibody or antigen-binding fragment thereof directed against a secreted S. aureus toxin. The combination of antibodies or fragments then binds to the S. aureus surface antigen and the secreted toxin to form two antigen-antibody complexes. In additional embodiments, the method includes contacting a test sample containing antigen-antibody complexes with at least one detectable label, wherein the signal produced by the detectable label(s) is directly correlated with the presence of S. aureus in the sample. For example, the detectable label(s) may include one or more fluorescent markers that bind an antibody or antigen in at least one of the antibody-antigen complexes such that an increase in fluorescence correlates with an increased concentration of S. aureus and/or secreted toxin in the sample .

[00185] In other embodiments, the at least one detectable label competes with a S. aureus surface antigen and/or secreted toxin for binding to the antibody or fragment combination. The signal produced by the detectable label(s) thus indirectly correlates with the concentration of S. aureus in the sample. For example, the detectable label(s) may include one or more fluorescent markers that compete with the surface antigen and/or secreted toxin for binding to the antibody such that a decrease in fluorescence correlates with an increased concentration of S. aureus and/or secreted toxin in the sample.

[00186] In certain embodiments, the detectable signals produced by the detectable labels in the test sample are compared with the signal from at least one control sample with known concentrations of antigens and antibodies. In embodiments using control samples, antibody-antigen complexes are detected in the control and test samples using detectable labels, and any statistically significant difference in detected signals among samples indicates the concentration, presence or absence of S. aureus and/or secreted toxin in the test sample.

[00187] In certain embodiments, the detection method is used to detect the presence of S. aureus in a patient sample, and the method further includes diagnosing the patient with an S. aureus infection. In some embodiments, the method is adapted for use in an automatic or semi-automatic system.

[00188] Certain embodiments also provide kits containing at least one antibody or antigen-binding fragment disclosed herein that are useful for various research and diagnostic purposes. For example, the kits can be used to detect S. aureus in a sample or to immunoprecipitate secreted S. aureus toxin. For isolation and purification purposes, the kit may contain an antibody or antigen binding fragment thereof bound to a bead (eg, Sepharose beads).

[00189] In this application, the use of the singular form includes the plural form unless specifically stated otherwise. Also in this application, the use of “or” means “and/or” unless otherwise indicated. Moreover, the use of the expression “including” as well as other forms such as “includes” and “included” is not limiting. Any range described herein should be understood to include the endpoints and all values between the endpoints.

[00190] The section headings used herein are for organization purposes only and should not be construed as limiting the subject matter described. All documents or portions of documents referred to herein, including without limitation patents, patent applications, articles, books and monographs, are therefore expressly incorporated herein by reference in their entirety for all purposes. To the extent that publications and patents or patent applications incorporated by reference are contrary to the present invention set forth in this specification, to that extent the present specification will control any contrary material.

Examples

[00191] The following examples serve to illustrate and in no way limit the present disclosure.

Example 1 - Materials and Methods

[00192] The following are the materials and methods used for Example 2 through Example 9.

Neutralization of hemolytic activity

[00193] Fifty microliters of each B cell hybridoma culture supernatant was mixed with recombinant alpha-toxin-His (rAT-his, with a final concentration of 0.1 μg/ml) in 96-well plates, followed by the addition of 50 μl of 5% red blood cells (RBC) rabbit in PBS. Control wells contained only RBC and culture media with or without AT. The plates were incubated for 1 hour at 37°C, and intact cells were pelleted by centrifugation. 50 μl of supernatants were transferred to a new 96-well plate and A490 was measured in a spectrophotometer. Neutralizing activity was calculated relative to lysis with RBC and rAT-his separately as follows: % inhibition = 100×[100-(A ₄₉₀ nAT+ Ab) /(A ₄₉₀ nAT without Ab)].

[00194] Inhibition of purified mAbs was also tested. Anti-AT MAb was added to a 96-well plate at 80 μg/ml in PBS, and samples were serially diluted (2-fold) in PBS to a final volume of 50 μl. Nonspecific IgG1 (R347) was included as an isotopic control. Twenty-five microliters of mAb dilutions were mixed with 25 μl of nAT (native alpha toxin) at a concentration of 0.1 μg/ml in 96-well round-bottom plates, followed by the addition of 50 μl of 5% RBC. Inhibition of hemolytic activity was calculated as described above.

Neutralization of A549 lysis

[00195] A549 cells were maintained in a 5% CO ₂ 37°C incubator in RMPI supplemented with the non-essential amino acid glutamine and 10% fetal bovine serum. Cells were washed once with Henck's balanced medium and seeded at 10 ⁴ /well in 50 μl RPMI, 5% FBS, and incubated at 37°C with 5% CO ₂ for 20 hours. Anti-AT MAb was added to a 96-well plate at a concentration of 80 μg/ml in RPMI and samples were serially diluted (2-fold) in RPMI. The mismatched IgG1 (R347) was included as an isotopic control. In a separate 96-well plate, 30 μl of diluted antibodies was mixed with 30 μl of nAT (for a final concentration of 5 μg/ml). Fifty microliters from each well was transferred to a plate containing adherent A549 cells. Control wells of A549 cells with or without nAT were included. The plates were incubated at 37°C with 5% CO ₂ for 3 hours, centrifuged and 50 μl of the supernatant was transferred to a new 96-well plate. Cell lysis was measured as lactate dehydrogenase (LDH) release using a Cytotox 96 non-radioactive assay kit (Promega) following the manufacturer's protocol. Background LDH was subtracted from each well and inhibition of LDH release was calculated: % inhibition = 100×[100-(A ₅₉₀ nAT+ Ab)/(A ₅₉₀ nAT without Ab)].

Neutralization of THP-1 lysis

[00196] THP-1 cells were maintained in a 5% CO ₂ , 37°C incubator in RPMI medium (Invitrogen) supplemented with non-essential amino acids (Invitrogen), 2 mM glutamine (Invitrogen) and 10% fetal bovine serum (Invitrogen). Anti-AT MAb was added to a 96-well plate at approximately 80 μg/mL in RPMI and samples were serially diluted (2-fold) in RPMI to a final volume of 50 μL. The mismatched IgG1 (R347) was included as an isotopic control. Twenty-five microliters of mAb dilutions were mixed with 25 μl of native alpha toxin (nAT) at a final concentration of 1.5 μg/ml, followed by the addition of 50 μl of THP-1 cells washed with RMPI (10 ⁶ cells/ml in RPMI with 10% FBS) in a 96-well plate. Control wells contained only THP-1 cells or with nAT. The plates were incubated in 5% CO ₂ at 37°C in an incubator for 3 hours, centrifuged, and 50 μl of the supernatant was transferred to a new 96-well plate. Cell lysis was measured as lactate dehydrogenase (LDH) release using a Cytotox 96 non-radioactive assay kit (Promega) following the manufacturer's protocol. Inhibition of LDH release was calculated as described above.

Mouse model of pneumonia

[00197] Twenty-four hours before infection, a group of ten 7-9 week old C57BL/6J mice (Harlan) received 0.5 ml of mAb at the indicated concentrations via intraperitoneal injection. The animals were then anesthetized with isoflurane, held upright, and the left and right nostrils inoculated with 0.05 ml of a bacterial suspension of S. aureus (1 x 10 ⁸ CFU - 3 x 10 ⁸ CFU) in sterile PBS. Animals were caged in the supine position for recovery and observed twice daily during the course of the study. Animal survival was monitored for a maximum of 6 days.

[00198] Alternatively, the animals were euthanized by CO ₂ inhalation 48 hours after bacterial infection. Lung and kidney were collected in sterile PBS, homogenized, diluted, and plated for bacterial enumeration. Statistical significance of mortality studies was determined using the log-rank test. The significance of bacterial isolation from organs was calculated using analysis of variance and Dunnett's post hoc test.

Mouse model of dermatotonecrosis

[00199] Groups of five 6-8 week old female BALB/c mice (Harlan) had their backs shaved and given an intraperitoneal injection of 0.5 ml IgG at the concentration shown in the graph. Twenty-four hours later, mice were infected by subcutaneous injection of 50 μl of bacterial suspension (1×10 ⁸ S. aureus). Animals were monitored twice daily for signs of infection and abscess size measured at the same time daily. The area of affected areas was calculated using the formula A=L×W. Statistical significance was determined using analysis of variance and Dunnett's post hoc test.

Mouse model of sepsis

[00200] Bacterial Challenge Dose Preparation: S. (USA300) aureus strain SF8300 was obtained from Binh Diep (University of California, San Francisco). Bacteria were cultured overnight at 37°C in 50 ml tryptic soy broth (TSB) with stirring at 250 rpm. Ten ml from a 24-hour culture was added to 1 liter of fresh TSB, and the bacteria were grown at 37°C with stirring to an optical density of 0.8 at 600 nm (OD600). Bacteria were recovered by centrifugation at 8000 rpm. within 15 min. at 4°C and washed in phosphate-buffered saline (PBS). Bacteria were collected by centrifugation and resuspended in PBS with 10% glycerol to a final bacterial stock concentration of ~2 x ¹⁰ CFU/ml.

[00201] Mice challenge and survival: BALB/c mice in groups of ten females, 8-9 weeks old, were injected intraperitoneally (IP) with LC10 at the indicated concentrations or mAb R347 (45 mg/kg) in 500 μl PBS. Animals were challenged 24 hours later by intravenous (IV) tail vein administration of 200 μl of a bacterial suspension (5 x 10 ⁷ CFU, diluted in PBS, pH 7.2, from a frozen stock solution). Survival of mice was monitored for 14 days after challenge. Statistical analysis was performed using the log-rank test: animals immunized with R347 (control) versus animals immunized with LC10 (Ab to Ab).

[00202] Bacterial load in the heart: infected mice were killed with CO2 14 hours after infection. The heart was removed, homogenized in tubes containing Lysis Matrix A in 1 ml of cold PBS, and plated on TSA plates for bacterial enumeration. Bacterial load in cardiac tissue was analyzed by pairwise comparison of mAbs R347 and LC10 using an unpaired two-tailed Student's t test. Data were considered significant if p<0.05.

[00203] Bacterial load in the blood: animals were killed with CO2 at 8, 24, 48, 72 and 144 hours after infection. Blood was collected by cardiac puncture and 100 μl was immediately plated on a TSB plate for CFU counting. Data were analyzed using an unpaired Student's t test. Values for mAbs LC10 and R347 were considered statistically different if p<0.05.

Receptor binding assay

[00204] Red blood cell “shadows” were prepared by incubating 5 ml of washed and packaged rabbit red blood cells (RBC) in 500 ml of lysis buffer (5 mM phosphate, 1 mM EDTA, pH 7.4) overnight at 4°C C with constant stirring. The shadows were then removed by centrifugation at 15,000×g and washed with 3× lysis buffer. They were then washed in PBS and resuspended in a final volume of 3 ml.

[00205] To assess the binding of nAT to shadow cell membranes, RBCs were diluted to an OD ₆₀₀ of approximately 0.2 in PBS and 50 μl was applied to ½-wells of 96-well plates (Costar) and incubated overnight at 4°C C. The liquid was then removed from the plates and the wells were blocked with 100 μl of 1% BSA in PBS, pH 7.4 for 2 hours at 4°C and washed 3x with PBS. A 20-molar excess of IgG was mixed with nAT at a concentration of 3 μg/ml and 50 μl was added to blocked plates. The plates were incubated at 4°C for 2 hours and washed 3× with PBS. Biotin-labeled rabbit anti-AT IgG was added to the wells at a concentration of 1 mg/ml and incubated at 4°C for 1 hour, washed 3×, and incubated with peroxidase-streptavidin conjugate (1:30000, Jackson Immunoresearch). Wells were washed 3x and developed with Sure Blue Reserve (KPL, Inc.). A ₄₅₀ was read using a plate reader (Molecular Devices) and % AT bound was calculated. % bound AT = 100×(A ₄₅₀ -AT+IgG/A ₄₅₀ - AT only).

Measurement of reaction and binding rate constants ( _KD )

[00206] Reaction rate constants (k _on , k _off ) for the binding of anti-Ab IgG antibodies to purified nAT were measured using the IgG capture assay format on a BIAcore 3000 instrument (BIAcore, Inc). Briefly, rat anti-mouse IgG was immobilized on a CM5 sensor chip according to the manufacturer's instructions. The final surface density of the capture reagent on the sensor chip was approximately 2500 response units (RU), which are described herein. A reference flow cell surface was also obtained on this sensor chip using an identical immobilization protocol and excluding nAT. IgG antibodies to AT were prepared at 20 nM in instrument buffer (HBS-EP buffer containing 0.01 M HEPES, pH 7.4, 0.15 M NaCl, 3 mM EDTA and 0.005% P-20), and two-fold serial dilutions of nAT. Serial dilutions of nAT were made ranging from approximately 0.78 nM to approximately 50 nM in instrument buffer.

[00207] A sequential approach was used for kinetic measurements. Each AT IgG was first injected onto the capture surface and reference surface at a flow rate of 50 μL/min. After binding of captured IgG had stabilized, a single concentration of nAT protein was injected onto both surfaces at a flow rate of 50 μL/min. The resulting binding reaction curves were used to obtain association phase data. Then, after nAT injection, the flow was switched back to instrument buffer for 10 minutes to allow dissociation phase data collection, followed by a 1 minute supply of 10 mM glycine, pH 1.5, to regenerate the IgG capture surface on the chip . Binding reactions resulting from repeated administrations of each nAT concentration were recorded for all anti-AT IgGs.

[00208] In addition, several injections of the buffer were made randomly during the series of injections. Selected buffer injections were used together with reference cell responses to correct raw data sets for injection artifacts and/or interactions caused by nonspecific binding, commonly referred to as “double control” (DG Myszka, Improving biosensor analysis. J. Mol. Recognit. 12 (1999) ), pp. 279-284). The fully corrected binding data were then generally fit 1:1 to a binding model (BIAevaluation 4.1 software, BIAcore, Inc, Uppsala, Sweden) with a parameter included to correct for mass transfer-limited binding to be detected. Using these analyses, reaction rate constants (on, off) were determined, from which the apparent K _D was then calculated as k _off /k _on .

Measuring cytokine levels in S. aureus-infected lungs

[00209] Seven to nine week old C57BL/6J mice were treated with 2A3.1hu (fully human 2A3.1) or R347 (45 mg/kg) by intraperitoneal injection 24 hours before intranasal infection with 1.5 x 10 ⁸ CFU USA300 (BAA- 1556, ATCC). Four and twenty-four hours after infection, mice were euthanized and the lungs were washed 3× with 1 ml of PBS. Bronchoalveolar lavage fluid (BAL) was stored at -70°C. Proinflammatory cytokines were assessed using the 7 pro-inflammatory II mouse cytokine kit (Mesoscale, Gaithersburg, MD) according to the manufacturer's instructions. Cytokine levels were expressed as pg/ml.

Dot blot tests

[00210] Overlapping peptides spanning amino acids 40-293 were chemically synthesized (New England Peptide). An attempt was made to synthesize AT _1-50 , but without success. Alpha toxin (AT), AT peptides, and AT fragments (1 μg) were spotted onto nitrocellulose and blocked for 10 min with casein blocking in PBS. The blots were then probed with 2 μg/ml single IgG for 3 hours at room temperature. Blots were washed and incubated with alkaline phosphatase conjugated to goat anti-mouse IgG or goat anti-rabbit IgG (1:1000, Caltag Laboratories) for 1 hour and developed using the BCIP/NBT Membrane Phosphatase Substrate System (KPL, Inc. ).

Example 2 - Selection and verification of targets

[00211] Thirteen surface antigens and four secreted toxins were selected for screening as antibody targets based on their conservation among clinical isolates and/or published vaccine activity. This group included alpha toxin and three soluble modulins (PSMs). Also included were 8 staphylococcal cell wall-anchored antigens/adhesins. Five of the selected targets had homologues in S. aureus and S. epidermidis. These targets are involved in nutrient absorption, biofilm formation, and cell division. Antibodies to alpha toxin have been used as a proposed way to reduce or neutralize toxin activities such as tissue damage and immune system dysregulation. Also used as targets were S. aureus surface determinants (IsdH, SdrC, ClfB, ClfA, and IsdB), which are important for S. aureus colonization, immune evasion, and fitness. A potential approach being considered to improve antibody therapy has included combining opsonic and toxin-neutralizing monoclonal antibodies.

[00212] Antibodies raised against these targets are collected in both in vitro and in vivo assays to reduce virulence and/or reduce colonization and immune evasion. Targeted fitness through active/passive immunization in murine models of infection was also assessed.

Example 3 - Detection of antibodies to IsdH

[00213] The primary target identified was iron-regulated surface determinant H (IsdH) of S. aureus. IsdH contains a 7-amino acid loop between the B1b and B2 β-sheets, and this 7-amino acid loop is conserved among several members of the iron-regulated surface determinant family, including IsdA and IsdB. Mutations in this 7-amino acid loop reduce the ability of S. aureus to bind hemoglobin by more than 100-fold and also impair the ability of S. aureus to evade phagocytic killing. Visai et al., J. Microbiology, 155(3): 667–679 (2008).

[00214] Anti-IsdH monoclonal antibodies (mAB) were detected using VelocImmune® mice (Regeneron Pharmaceuticals) and phage panning (Dyax or CAT libraries). Fifty-nine IgG antibodies were purified (29 from Dyax libraries, 16 from CAT libraries, and 14 from VelocImmune mice).

Example 4 - mAB to IsdH screening cascade

[00215] Identified anti-IsdH mABs were assessed using an in vitro S. aureus whole-cell binding ELISA. Antibodies were also screened by ELISA for inhibition of S. aureus haptoglobin binding. Antibodies were then assessed in an opsonophagocytic killing (OPK) assay (described below). Eleven IgG antibodies to IsdH were detected, which had opsonic activity against 4 S. aureus isolates. Five anti-IsdH antibodies efficiently bound S. aureus after in vivo seeding in a mouse model of infection (described below). These five best anti-IsdH mABs (3 from the Dyax libraries and 2 from the CAT libraries) were selected for industrial antibody production, affinity testing, and subsequent in vivo testing. In vivo testing included studies in a bacteremia model (described below). Antibody 2F4 significantly reduced CFU in a bacteremia model. The five antibodies were then characterized and evaluated for use in combination therapy.

[00216] The opsonophagocytic killing (OPK) assay involved combining 10 μl of S. aureus (10 ⁶ cells/ml), 10 μl of monoclonal antibody and 60 μl of DMEM plus 0.1% gelatin. The solution was incubated for 30 minutes at 4°C. After 30 minutes, 10 μl of human promyelocytic leukemia cells (HL-60) was added at a rate of 10 ⁷ cells/ml along with 10 μl of human serum pre-absorbed for S. aureus. At time _T0 , 10 μl of the solution was seeded and then incubated at 37°C with stirring at 1500 rpm. within 60 minutes. At time _T60 , HL-60 cells were lysed with 1% saponin, replated, and CFU concentration determined. The percentage of OPK was calculated as follows: 100×(1-(T ₆₀ /T ₀ )), where T ₆₀ refers to the CFU concentration at the end of the assay (i.e., after 60 minutes), and T ₀ refers to the CFU concentration at beginning of the analysis.

[00217] Eleven anti-IsdH monoclonal antibodies were identified that had sufficient opsonic activity against 4 S. aureus isolates to warrant additional testing. Figure 19 illustrates that antibodies B11, 2F4 and A7 had an increased percentage of OPK compared to the control antibody R347 when tested against S. aureus strains Newman and USA300.

[00218] To determine whether antigens that were targeted by antibodies in S. aureus were expressed in vivo, antibody binding was assessed after in vivo seeding in a mouse. Mice were challenged intraperitoneally with approximately 5x10 ⁸ CFU of S. aureus. After 1-6 hours, mice were exsanguinated and the blood pooled in ice-cold citrate. Eukaryotic cells were lysed with 1% NP-40. Lysed cells were washed three times with phosphate-buffered saline (PBS) and sonicated, followed by resuspension of S. aureus bacteria in buffer (approximately 0.5-10×10 ⁶ CFU recovered after lysis and resuspension). Antibodies to IsdH were introduced into cell lysates, and antibody binding was assessed by staining and the results of FACs sorting.

[00219] Five of the eleven anti-IsdH mABs (designated 1C1, 2F4, A7, IsdH003, and IsdH0016) bound S. aureus after in vivo seeding. Figure 2 shows the binding of antibodies B11, 2F4, A7 and 1C1 compared to the control antibody R347, in S. aureus strains ARC2081 and USA300. The figure shows that antibodies 2F4, A7 and 1C1 bound S. aureus ex vivo.

[00220] Two of the five anti-IsdH mABs (1C1 and 2F4) also competed with haptoglobin (Hp) for binding to IsdH, while the others did not. Figure 3 shows that antibody 1C1 competes with Hp for binding to the Neat-1 subunit on IsdH. Increasing 1C1 concentrations (in μg/ml) correlated with decreased Hp binding to IsdH compared to Hp binding in the presence of the control antibody R347. Likewise, Figure 3 shows that antibody 2F4 competes with Hp for binding to the Neat-2 subunit on IsdH. Increasing 2F4 concentrations (in μg/ml) correlated with decreased Hp binding to IsdH compared to Hp binding in the presence of the control antibody R347.

[00221] To evaluate whether antibodies 1C1, 2F4, A7, IsdH003 and IsdH0016 were effective when administered in vivo, a mouse model of bacteremia was used. Mice were injected intraperitoneally with 45, 15, or 5 mg/kg monoclonal antibody and then allowed to recover overnight. The next day, mice were injected intraperitoneally with approximately 10 ⁸ CFU of S. aureus (strain Newman). After approximately 4 hours, the blood was assessed for CFU concentration, measured as log[CFU/ml]. Figure 4 shows that antibodies 1C1, A7, IsdH003 and IsdH0016 did not reduce the concentration of CFU in the bacteremia model. However, Figure 5 shows that antibody 2F4 did reduce CFU concentrations in a mouse model of bacteremia.

[00222] Antibody 2F4 was further evaluated for ex vivo binding to various strains of S. aureus. The antibody bound to 23 of 25 S. aureus isolates after in vivo seeding and isolation in mice. Figure 6 illustrates 2F4 binding in strains ARC2379 (USA100), ARC2081 (USA200) and BAA-1556 (USA 300). The table below illustrates the results of binding experiments after in vivo seeding of 25 strains of S. aureus in a mouse.

Binding of antibody 2F4 after in vivo seeding of S. aureus strains

[00223] Antibody 2F4 was also evaluated in ORK assays involving S. aureus clinical isolates Newman, ARC634 (USA100), ARC2081 (USA200), and BAA-1556 (USA300). Figure 7 shows that 2F4 had opsonic activity against major clinical isolates of S. aureus.

[00224] 2F4 was then assessed for its affinity for IsdH and for the Neat-2 subunit in an lgGFc hu uptake assay. From the average affinity averaged over three experiments, the K _D for IsdH was found to be 3.66 nM and the K _D for Neat-2 was equal to 2.57 nM. Figure 8.

Example 5 - Combination therapy with anti-IsdH and anti-AT antibodies

[00225] Alpha toxin and IsdH play distinct roles during pathogenesis following S. aureus infection. The former is a secreted toxin, while the latter is a surface protein important for colonization, immune evasion, and bacterial fitness. Both may be differentially expressed during S. aureus infection. Combining monoclonal antibodies with different modes of action could potentially produce additive or synergistic effects while reducing the risk of a strain evading therapy.

[00226] 2A3, an anti-alpha toxin antibody, was evaluated for use in combination with 2F4, an anti-IsdH antibody. When administered in combination, antibodies 2A3 and 2F4 demonstrated synergistic effects in an organ loading model. Figure 9 shows that the kidney distribution of S. aureus strain USA300 was reduced in the presence of both antibodies compared to either antibody alone or compared to the control antibody R347.

[00227] The results of these combination therapy experiments suggested that a combination approach to the prevention or treatment of S. aureus may be effective.

Example 6 -mAbs to ClfA inhibited ClfA binding

[00228] mAbs to ClfA inhibited the binding of ClfA to immobilized fibrinogen in vitro. It was reported that ClfA as a virulence factor promoted the binding of S. aureus to fibrinogen present in blood plasma. This resulted in agglutination of bacteria in the blood.

[00229] The ability of three anti-ClfA mAbs generated using B-cell hybridoma technology to inhibit the binding of ClfA to immobilized fibrinogen was assessed. Antibody R347 was used as a negative control. The activity of each ClfA mAb in this assay was calculated at the IC50, the concentration required to initiate 50% inhibition of binding. As shown in Figure 10, in addition to the IC50 of each antibody, anti-ClfA antibodies inhibited the binding of ClfA to immobilized fibrinogen.

Example 7 - mAb 11H10 to ClfA inhibited agglutination of S. aureus in human plasma with three different clinical isolates.

[00230] To assess the agglutination of S. aureus in human plasma, bacteria were incubated with each anti-ClfA mAb and bacterial aggregation was visually examined after a 3-minute incubation at 37°C. To more accurately compare the activity of the mAbs in this assay, they were compared at the minimum concentration required to inhibit agglutination. 11H10 was more effective than 27H4 or 23D6 (Figure 11). Additionally, agglutination experiments were performed with three different clinical isolates, and 11H10 showed inhibition against these three isolates compared to two other anti-ClfA mAbs that covered one or two strains.

Example 8 - Binding to the 11H10 Epitope

[00231] Due to the different characteristics of 11H10 compared to 23D6 and 27H4, as discussed previously, its binding characteristics were further examined. Epitope competitive binding was performed using the Octet system to evaluate whether 11H10 binds to a different epitope than 23D6 and 27H4. As seen in Figure 12, 23D6 and 27H4 competed for binding to ClfA, suggesting that they might share a common binding site on ClfA. However, there was no competition between 11H10 and 23D6 or 11H10 and 27H4, demonstrating that the 11H10 epitope on ClfA was different compared to 23D6 and 27H4.

Example 9 - Passive immunization with mAb 11H10 to ClfA demonstrated effectiveness in a lethal IV challenge model

Bacterial load in the heart

[00232] To test whether agglutination of staphylococci occurred in vivo, mice were first infected in the tail vein with isolate USA300 and the number of bacteria in the heart was recorded 14 hours after infection. As shown in Figure 13, prophylactic intraperitoneal administration of the anti-ClfA 11H10 mAb (IP) at 45 mg/kg resulted in a significant reduction in bacterial cfu in the heart (p=0.031). It was dose dependent, as 11H10 at 15 mg/kg only slightly reduced the bacterial load in the heart compared to the negative control R347.

Survival

[00233] A sublethal dose of USA300 for IV challenge was determined to induce 20% survival after 2 weeks. In this model, the ability of mAb 11H10 to ClfA to increase animal survival was studied. Figure 14 shows that 11H10 injection resulted in a significant increase in survival (p=0.0114 at 45 mg/kg and p=0.0239 at 15 mg/kg) 2 weeks after infection.

Example 10 - Efficacy of the combination of mAb 11H10 to ClfA and Ab LC10 to AT in a model of lethal IV infection

[00234] Six-week-old female BALB/c mice were passively immunized intraperitoneally (IP) with mAb at the indicated concentrations (diluted in 500 μl PBS) and 24 hours later challenged intravenously (IV) with an LD20 tail vein dose of bacteria (in 200 μl PBS). Survival was monitored up to 14 days postinfection.

[00235] Data were analyzed using the log-rank test (Mantel-Cox), and the p value was considered statistically significant if ≤0.05. To test whether agglutination of staphylococci occurred in vivo, mice were first infected into the tail vein with isolate CA-MRSA USA300, HA-MRSA-100, or HA-MSSA USA200, and bacterial counts in the heart and kidney were recorded 14 hours after infection.

[00236] The effectiveness of the combination of mAb 11H10 to ClfA and Ab LC10 to AT was tested in a lethal IV infection model. As shown in Figure 15, prophylactic administration of anti-ClfA mAb 11H10, anti-AT mAb LC10, and a combination of both anti-ClfA mAb and anti-AT mAb LC10 resulted in a significant reduction in cfu of bacteria in the heart (Fig. 15b) and kidney (15c). .

[00237] Figure 15 also demonstrates the ability of anti-ClfA mAb 11H10, anti-AT mAb LC10, and the combination of anti-ClfA mAb 11H10 and anti-At mAb LC10 to improve animal survival in a mouse model of IV challenge using a sublethal dose of USA300. Figure 15a shows that the combination of both antibodies resulted in a significant increase in survival response 2 weeks post-infection compared to control and compared to any single anti-ClfA mAb and anti-AT mAb.

[00238] Figures 16 and 17 further demonstrate the ability of the combination of mAb 11H10 to ClfA and mAb LC10 to At to increase animal survival in a study in the IV infection model using a sublethal dose of HA-MRSA USA100 (Figure 16) and a sublethal dose of HA-MSSA USA200 (figure 17).

Example 11 - Efficacy of the combination of mAb 2F4 to IsdH and Ab LC10 to AT in a model of lethal IV infection

[00239] The experiments were carried out as described above for Example 10.

[00240] The effectiveness of the combination of mAb 2F4 to IsdH and Ab LC10 to AT was tested in a lethal IV infection model. As shown in Figure 18, the combination of anti-IsdH mAb 2F4 and anti-At mAb LC10 increased animal survival in an IV challenge model using a sublethal dose of HA-MRSA USA100. Figure 18 shows that the combination resulted in a significant increase in survival response 6 days post-infection compared to the R347 control.

Sequence table

Table 1
VL CDR sequences for mAbs 2A3.1, 10A7.5, 12B8.19 and 25E9.1 SEQ ID NO: Description Subsequence SEQ ID NO: 1 CDR1 VL RASQSISSWLA SEQ ID NO: 2 CDR2 VL KASSLES SEQ ID NO: 3 CDR3 VL QQYNSYWT

table 2
CDR VL sequences for mAB 28F6.1 SEQ ID NO: Description Subsequence SEQ ID NO: 4 CDR1 VL mAb 28F6.1 RASQGIRNDLG SEQ ID NO: 5 CDR2 VL mAb 28F6.1 DASSLQS SEQ ID NO: 6 CDR3 VL mAb 28F6.1 LQDYNYPWT

Table 3
CDR VH sequences for mAb 2A3.1 SEQ ID NO: Description Subsequence SEQ ID NO: 7 CDR1 VH SYDMH SEQ ID NO: 8 CDR2 VH GIGTAGDTYYPGSVKG SEQ ID NO: 9 CDR3 VH DNYSSTGGYYGMDV

Table 4
CDR VH sequences for mAbs 10A7.5 and 12B8.19 SEQ ID NO: Description Subsequence SEQ ID NO: 10 CDR1 VH RYDMH SEQ ID NO: 11 CDR2 VH VIGTDGDTYYPGSVKG SEQ ID NO: 12 CDR3 VH DRYSSSNHYNGMDV

Table 5
CDR VH sequences for mAb 28F6.1 SEQ ID NO: Description Subsequence SEQ ID NO: 13 CDR1 VH mAb 28F6.1 SYAMT SEQ ID NO: 14 CDR2 VH mAb 28F6.1 VISGSGGSTYYADSVKG SEQ ID NO: 15 CDR3 VH mAb 28F6.1 DGRQVEDYYYYYGMDV

Table 6.
CDR VH sequences for mAb 25E9.1 SEQ ID NO: Description Subsequence SEQ ID NO: 7 CDR1 VH mAb 25E9.1 SYDMH SEQ ID NO: 17 CDR2 VH mAb 25E9.1 VIDTAGDTYYPGSVKG SEQ ID NO: 18 CDR3 VH mAb 25E9.1 DRYSGNFHYNGMDV

Table 7
VL and VH amino acid sequences for alpha-toxin mAb Description VH or VL sequence (with CDRs in bold) CDR1 CDR2 CDR3 VL mAb 2A3.1 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GV
PSRFSGSGSGTEFTLT
ISSLQPDDFATYYCQ
QYNSYWT FGQGTK
V.E.I.K.
(SEQ ID NO: 19) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYNS
Y.W.T.
(SEQ ID NO: 3) VH mAb 2A3.1 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATG
KGLEWVS GIGTAGD
TYYPGSVKG RFTISR
ENAKNSLYLQLNSL
RAGDTAVYFCAR DN
YSSTGGYYGMDV W
GQGTTVTVSS
(SEQ ID NO: 20) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DNYSS
TGGYY
GMDV
(SEQ ID NO: 9) VL mAb 10A7.5 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GV
PSRFSGSGSGTEFTLT
ISSLQPDDFATYYCQ
QYNSYWT FGQGTK
V.E.I.K.
(SEQ ID NO: 21) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYNS
Y.W.T.
(SEQ ID NO: 3) VH mAb 10A7.5 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
SRYDMH WVRQATG
KGLEWVS VIGTDGD
TYYPGSVKG RFIISR
ENAKNSLYLEMNSL
RAGDTAVYYCAR D
RYSSSNHYNGMDV
WGQGTTVTVSS
(SEQ ID NO: 22) RYDMH
(SEQ ID NO: 10) VIGT
DGDT
YYPG
SVKG
(SEQ ID NO: 11) DRYSS
SNHYN
GMDV
(SEQ ID NO: 12) VL mAb 12B8.19 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKVLIY KASSLES GV
PSRFSGSGSGTEFTLTI
SSLQPDDFATYYC QQ
YNSYWT FGQGTKVEIK
(SEQ ID NO: 23) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYNS
Y.W.T.
(SEQ ID NO: 3) VH mAb 12B8.19 EVQLVESGGGLV
QPGGSLRLSCAAS
GFTFS RYDMH WV
RQATGKGLEWVS V
IGTDGDTYYPGSV
KG RFIISRENAKNSL
YLEMNSLRAGDTAV
YYCAR DRYSSSNHY
NGMDV WGQGTTVT
VSS
(SEQ ID NO: 24) RYDMH
(SEQ ID NO: 10) VIGT
DGDT
YYPG
SVKG
(SEQ ID NO: 11) DRYSS
SNHYN
GMDV
(SEQ ID NO: 12) VL mAb 28F6.1 AIQMTQSPSSLSAS
VGDRVTITC RASQ
GIRNDLG WYQQK
PGKAPKLLIY DASS
LQS GVPSRFSGSGS
GTDFTLTISSLQPED
FATYYC LQDYNYP
WT FGQGTKVEIK
(SEQ ID NO: 25) RASQGIRNDLG (SEQ ID NO: 4) DASSLQS
(SEQ ID NO: 5) LQDYN
YPWT
(SEQ ID NO: 6) VH mAb 28F6.1 EVQLLESGGGLVQP
GGSLRLSCAASGFT
FS SYAMT WVRQAP
GKGLEWVS VISGSG
GSTYYADSVKG RFT
VSRDNSKNTLYLQM
NSLRAEDTAVYYCA
K DGRQVEDYYYYY
GMDV WGQGTTVTVSS
(SEQ ID NO: 26) SYAMT
(SEQ ID NO: 13) VISGS
GGST
YYAD
SVKG
(SEQ ID NO: 14) DGRQV
EDYYY
YYGMDV
(SEQ ID NO: 15) VL mAb 25E9.1 DIQMTQSPSTLSASV
GDRVTTITC RASQSISS
WLA WYQQKPGKAPK
LLIY KASSLES GVPSR
FSGSGSGTEFTLTISSL
QPDDFATYYC QQYN
SYWT FGQGTKVEIK
(SEQ ID NO: 27) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYNS
Y.W.T.
(SEQ ID NO: 3) VH mAb 25E9.1 EVQLVESGGGLVQPG
GSLRLSCTASGFTFS SY
DMH WVRQATGKGLE
WV SVIDTAGDTYYPG
SVKG RFTISRENAKNSL
YLQMNSLRAGDTAVYY
CVR DRYSGNFHYNGM
DV WGQGTTVTVSS
(SEQ ID NO: 28) SYDMH
(SEQ ID NO: 7) SVID
TAGD
TYYP
GSVKG
(SEQ ID NO: 17) DRYSG
NFHYN
GMDV
(SEQ ID NO: 18) VH mAb QD20 EVQLVESGGGLVQPG
GSLRLSCAASGFTFS S
YDMH WVRQATGKGL
EWVS GIGTAGDTYY
PGSVKG RFTISRENAK
NSLYLQMNSLRAGDT
AVYYCAR DRYSPTG
HYMGMDV WGQGTT
VTVSS
(SEQ ID NO: 41) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DRYSP
TGHYM
GMDV
(SEQ ID NO
: 16) VL mAb QD20 DIQMTQSPSTLSASVG
DRVTITC RASQSISS
WLA WYQQKPGKA
PKLLIY KASSLES G
VPSRFSGSGSGTEFT
LTISSLQPDDFATYY
C QQYDTYWT FGQG
TKVEIK
(SEQ ID NO: 42) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYDT
Y.W.T.
(SEQ ID NO: 64) VH mAb QD33 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATG
KGLEWVS GIGTAGD
TYYPGSVKG RFTISR
ENAKNSLYLQMNSL
RAGDTAVYYCAR DR
YSRTGHYMGMDV W
GQGTTVTVSS
(SEQ ID NO: 43) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DRYSR
TGHYM
GMDV
(SEQ ID NO: 65) VL mAb QD33 DIQMTQSPSTLSASVG
DRVTITC RASQSISSW
LA WYQQKPGKAPKL
LIY KASSLES GVPSRF
SSGSGSGTEFTLTISSLQ
PDDFATYYC QQYDTY
WT FGQGTKVEIK
(SEQ ID NO: 44) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYDT
Y.W.T.
(SEQ ID NO: 64) VH mAb QD37 EVQLVESGGGLVQPG
GSLRLSCAASGFTFS SY
DMH WVRQATGKGLE
WVS GIGTAGDTYYP
GSVKG RFTISRENAKN
SLYLQMNSLRAGDTA
VYYCAR DRYSRTG
HYMGMSL WGQGTT
VTVSS
(SEQ ID NO: 45) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DRYSR
TGHY
MGMSL
(SEQ ID NO
: 66) VL mAb QD37 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GVP
SRFSGSGSGTEFTLTIS
SLQPDDFATYYC QQY
DTYWT FGQGTKVEIK
(SEQ ID NO: 46) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYDTY
WT(SEQ ID NO: 64) VH mAb QD3 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATG
KGLEWVS GIGTAGD
TYYPGSVKG RFTISR
ENAKNSLYLQMNSL
RAGDTAVYYCAR DN
YSRTGHYMGMDV W
GQGTTVTVSS
(SEQ ID NO: 47) SYDMH
(SEQ ID NO: 7) GIGTA
GDTY
YPGSV
KG
(SEQ ID NO: 8) DNYSR
TGHYM
GMDV
(SEQ ID NO: 67) VL mAb QD3 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GV
PSRFSGSGSGTEFTLT
ISSLQPDDFATYYC K
QYADYWT FGQGTK
V.E.I.K.
(SEQ ID NO: 48) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) KQYAD
Y.W.T.
(SEQ ID NO: 68) VH mAb QD4 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATG
KGLEWVS GIGTAGD
TYYPGSVKG RFTISR
ENAKNSLYLQMNSLR
AGDTAVYYCAR DNY
SRTGHYMGMDV WG
QGTTVTVSS
(SEQ ID NO: 49) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DNYS
RTGH
YMGMDV
(SEQ ID NO: 67) VL mAb QD4 DIQMTQSPSTLSASVG
DRVTITC RASQSISSW
LA WYQQKPGKAPKL
LIY KASSLES GVPSRF
SSGSGSGTEFTLTISSLQ
PDDFATYYC QQYDTY
WT FGQGTKVEIK
(SEQ ID NO: 50) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYDT
Y.W.T.
(SEQ ID NO: 64) VH mAb QD23 EVQLVESGGGLVQ
PGGSLRLSCAASGF
TFS SYDMH WVRQA
TGKGLEWVS GIGTA
GDTYYPGSVKG RFT
ISRENAKNSLYLQM
NSLRAGDTAVYYCA
R DRYSPTGYMGM
SL WGQGTTVTVSS
(SEQ ID NO: 51) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DRYSP
TGHYM
GMSL
(SEQ ID NO: 78) VL mAb QD23 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GV
PSRFSGSGSGTEFTLT
ISSLQPDDFATYYC QQ
YDTYWT FGQGTKVEIK
(SEQ ID NO: 52) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYDT
Y.W.T.
(SEQ ID NO: 64) VH mAb QD32 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATGK
GLEWVS GIGTAGDT
YYPGSVKG RFTISRE
NAKNSLYLQMNSLRA
GDTAVYYCAR DRYS
RTGHYMGMDV WGQ
GTTVTVSS
(SEQ ID NO: 53) SYDMH
(SEQ ID NO: 7) GIGTA
GDTYY
PGSVKG
(SEQ ID NO: 8) DRYSR
TGHYM
GMDV
(SEQ ID NO: 65) VL mAb QD32 DIQMTQSPSTLSASVG
DRVTITC RASQSISSW
LA WYQQKPGKAPKL
LIY KASSLES GVPSRF
SSGSGSGTEFTLTISSLQ
PDDFATYYC KQYADY
WT FGQGTKVEIK
(SEQ ID NO: 54) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) KQYAD
Y.W.T.
(SEQ ID NO: 68) VH mAb 2A3GL EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SYDMH WVRQATG
KGLEWVS GIGTAG
DTYYPGSVKG RFTI
SRENAKNSLYLQMN
SLRAGDTAVYYCAR
DNYSSTGGYYGMDV
WGQGTTVTVSS
(SEQ ID NO: 55) SYDMH
(SEQ ID NO: 7) GIGTA
GDTY
YPGS
VKG
(SEQ ID NO: 8) DNYS
STGGY
YGMDV
(SEQ ID NO: 9) VL mAb 2A3GL DIQMTQSPSTLSAS
VGDRVTITC RASQSI
SSWLA WYQQKPGK
APKLLIY KASSLES G
VPSRFSGSGSGTEFTL
TISSLQPDDFATYYC Q
QYNSYWT FGQGT
KVEIK
(SEQ ID NO: 56) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) QQYNSYWT
(SEQ ID NO: 3) VH mAb LC10 EVQLVESGGGLVQP
GGSLRLSCAASGFTF
S SHDMH WVRQATG
KGLEWVS GIGTAGD
TYYPDSVKG RFTISR
ENAKNSLYLQMNSL
RAGDTAVYYCAR DR
YSPTGHYYGMDV W
GQGTTVTVSS
(SEQ ID NO: 57) SHDMH
(SEQ ID NO: 69) GIGT
AGDT
YYPD
SVKG
(SEQ ID NO: 70) DRYSPTGYYGMDV
(SEQ ID NO: 71) VL mAb LC10 DIQMTQSPSTLSASVG
DRVTITC RASQSISSW
LA WYQQKPGKAPKL
LIY KASSLES GVPSRF
SSGSGSGTEFTLTISSLQ
PDDFATYYC KQYAD
YWT FGQGTKVEIK
(SEQ ID NO: 58) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) KQYAD
Y.W.T.
(SEQ ID NO: 68) VH mAb TVES EVQLVESGGGLVQPG
GSLRLSCAASGFTFS S
YDMHW VRQATGKGL
EWVS GIGTAGDTYYP
GSVKG RFTISRENAKN
SLYLQMNSLRAGDTA
VYYCAR DNYSPTGG
YYGMDV WGQGTTV
TVSS
(SEQ ID NO: 59) SYDMH
(SEQ ID NO: 7) GIGT
AGDT
YYPG
SVKG
(SEQ ID NO: 8) DNYSP
TGGYY
GMDV
(SEQ ID NO: 72) VL mAb TVES DIQMTQSPSTLSAS
VGDRVTITC RASQ
SISSWLA WYQQKPG
KAPKLLIY KASSLKS
GVPSRFSGSGSGTEFT
LTISSLQPDDFATYYC
QQYESYWT FGQGT
KVEIK
(SEQ ID NO: 60) RASQSISSWLA
(SEQ ID NO: 1) KASSLKS
(SEQ ID NO: 73) QQYES
Y.W.T.
(SEQ ID NO: 74) VH mAb 3H7KAD EVQLVESGGGLV
QPGGSLRLSCAAS
GFTFS SHDMH WV
RQATGKGLEWVS G
IGTRGDTYYPDSV
KG RFTISRENAKNS
LYLQMNSLRAGDTA
VYYCAR DRYSPTG
HYYGMDV WGQG
TTVTVSS
(SEQ ID NO: 61) SHDMH
(SEQ ID NO: 69) GIGT
RGDT
YYPD
SVKG
(SEQ ID NO: 75) DRYSP
TGHYY
GMDV
(SEQ ID NO: 71) VL mAb 3H7KAD DIQMTQSPSTLSAS
VGDRVTITC RASQ
SISSWLA WYQQKP
GKAPKLLIY KASSL
ES GVPSRFSGSGSG
TEFTLTISSLQPDDF
ATYYC KQYADYW
T FGQGTKVEIK
(SEQ ID NO: 58) RASQSISSWLA
(SEQ ID NO: 1) KASSLES
(SEQ ID NO: 2) KQYAD
Y.W.T.
(SEQ ID NO: 68) VH mAb LC9 EVQLVESGGGLVQ
PGGSLRLSCAASGF
TFS SHDMH WVRQA
TGKGLEWVS GIGT
RGDTYYPDSVKG R
FTISRENAKNSLYLQ
MNSLRAGDTAVYY
CAR DKYSPTGHYY
GMDV WGQGTTVTVSS
(SEQ ID NO: 62) SHDMH
(SEQ ID NO: 69) GIGT
RGDT
YYPD
SVKG
(SEQ ID NO: 75) DKYSP
TGHYY
GMDV
(SEQ ID NO: 76) VL mAb LC9 DIQMTQSPSTLSASV
GDRVTITC RASQSIS
SWLA WYQQKPGKA
PKLLIY KASSLES GV
PSRFSGSGSGTEFTLT
ISSLQPDDFATYYC
KQYADYWT FGQG
TKVEIK
(SEQ ID NO: 58) RASQSISSWLA
(SEQ ID NO: 1) KASSL
ES
(SEQ ID NO: 2) KQYAD
Y.W.T.
(SEQ ID NO: 68) VH mAb LC4 EVQLVESGGGLV
QPGGSLRLSCAAS
GFTFS SHDMH WV
RQATGKGLEWVS
G IGTRGDTYYPDS
VKG RFTISRENAKN
SLYLQMNSLRAGD
TAVYYCAR DKYSP
TGHYYGMDV WG
QGTTVTVSS
(SEQ ID NO: 62) SHDMH
(SEQ ID NO: 69) GIGT
RGDT
YYPD
SVKG
(SEQ ID NO: 75) DKYSPTGYYGMDV
(SEQ ID NO: 76) VL mAb LC4 DIQMTQSPSTLSAS
VGDRVTTITC RASQS
ISSWLA WYQQKPG
KAPKLLIY KASSLV
K GVPSRFSGSGSGTE
FTLTISSLQPDDFATY
YC QQYESYWT FGQ
GTKVEIK
(SEQ ID NO: 63) RASQSISSWLA
(SEQ ID NO: 1) KASSLVK
(SEQ ID NO: 77) QQYESYWT
(SEQ ID NO: 74) VH mAb LC5 EVQLVESGGGLV
QPGGSLRLSCAASG
FTFS SHDMH WVRQ
ATGKGLEWVS GIGT
AGDTYYPDSVKG R
FTISRENAKNSLYLQ
MNSLRAGDTAVYY
CAR DRYSPTGYY
GMDV WGQGTTVTVSS
(SEQ ID NO: 79) SHDMH
(SEQ ID NO: 69) GIGTA
GDTY
YPDS
VKG
(SEQ ID NO: 70) DRYSP
TGHYY
GMDV
(SEQ ID NO: 71) VL mAb LC5 DIQMTQSPSTLSAS
VGDRVTTITC RASQS
ISSWLA WYQQKPG
KAPKLLIY KASSLV
K GVPSRFSGSGSGT
EFTLTISSLQPDDFA
TYYC QQYESYWT F
GQGTKVEIK
(SEQ ID NO: 63) RASQ
SISSW
L.A.
(SEQ ID NO: 1) KASS
LVK
(SEQ ID NO: 77) QQYES
Y.W.T.
(SEQ ID NO: 74)

Table 8
VL and VH nucleotide sequences for alpha-toxin mAb SEQ ID NO: Description Subsequence SEQ ID NO: 29 Nucleotide sequence of VL mAb 2A3.1 GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATAAGGCGTCTAGTTTAGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCA GCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 30 Nucleotide sequence of VH mAb 2A3.1 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTACGACATGCACTGGGTCCGCCAAGCTACAGGAAAAGGTCTGGAGTGGGTCTCAGGTATTGGCACTGCTGGTGACACATATTATCCAGGCTCCGTGAAGGGCCGATTCACCATCTCCAGAGAAAATGCCAAGAACTCCTTGTATCTT CAATTGAACAGCCTGAGAGCCGGGGACACGGCTGTGTACTTCTGTGCAAGAGACAATTATAGCAGCACCGGGGGGTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 31 Nucleotide sequence of VL mAb 10A7.5 GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATAAGGCGTCTAGTTTAGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCA GCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 32 Nucleotide sequence of VH mAb 10A7.5 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGGTACGACATGCACTGGGTCCGCCAAGCTACAGGAAAAGGTCTGGAGTGGGTCTCAGTTATTGGTACTGATGGTGACACATACTATCCAGGCTCCGTGAAGGGCCGATTCATCATCTCCAGAGAAAATGCCAAGAACTCCTTGTATCTT GAAATGAACAGCCTGAGAGCCGGGGACACGGCTGTGTATTACTGTGCAAGAGATCGGTATAGCAGCTCGAACCACTACAACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 33 Nucleotide sequence of VL mAb 12B8.19 GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGGTCCTGATCTATAAGGCGTCTAGTTTAGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTG CAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 34 Nucleotide sequence of VH mAb 12B8.19 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGGTACGACATGCACTGGGTCCGCCAAGCTACAGGAAAAGGTCTGGAGTGGGTCTCAGTTATTGGTACTGATGGTGACACATACTATCCAGGCTCCGTGAAGGGCCGATTCATCATCTCCAGAGAAAATGCCAAGAACTCCTTGTATCTT GAAATGAACAGCCTGAGAGCCGGGGACACGGCTGTGTATTACTGTGCAAGAGATCGGTATAGCAGCTCGAACCACTACAACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 35 Nucleotide sequence of VL mAb 28F6.1 GCCATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGAAATGATTTAGGCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCATCCAGTTTACAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGAATCTGGCACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCT GAAGATTTTGCAACTTATTACTGTCTACAAGATTACAATTACCCGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 36 Nucleotide sequence of VH mAb 28F6.1 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGGTCTCAGTTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCGTCTCCAGAGACAATTCCAAGAACACGCT GTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAGATGGGAGGCAGGTCGAGGATTACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA SEQ ID NO: 37 Nucleotide sequence of VL mAb 25E9.1 GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCGTCTAGTTTAGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTG CAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 38 Nucleotide sequence of VH mAb 25E9.1 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTCAGTAGTTACGACATGCACTGGGTCCGCCAAGCTACAGGAAAAGGTCTGGAGTGGGTCTCAGTTATTGATACTGCTGGTGACACATACTATCCAGGCTCCGTGAAGGGCCGATTCACCATCTCCAGAGAAAATGCCAAGAACTCCTTGTATCTTC AAATGAACAGCCTGAGAGCCGGGGACACGGCTGTGTATTACTGTGTAAGAGATAGGTATAGTGGGAACTTCCACTACAACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA

Table 9
Summary table of CDR VL and VH for alpha toxin Description SEQ ID NO CDR 1 VL 14 CDR 2 VL 2, 5, 73, 77 CDR 3 VL 3, 6, 64, 68, 74 CDR 1 VH 7, 10, 13, 69 CDR 2 VH 8, 11, 14, 17, 70, 75 CDR 3 VH 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76, 78

Table 10
Amino acid sequences of VL and VH mAbs to alpha toxin containing a variant Fc region SEQ ID NO: Description Subsequence SEQ ID NO: 130 LC 10-VH-IgG1-YTE: EVQLVESGGGLVQPGGSLRLSCAASGFTFSSHDMHWVRQATGKGLEWVSGIGTAGDTYYPDSVKGRFTISRENAKNSLYLQMNSLRAGDTAVYYCARDRYSPTGHYYGMDVWGQGTTVTVSS-ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 131 VL-kappa LC10 DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCKQYADYWTFGQGTKVEIK-RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK VYACEVTHQGLSSPVTKSFNRGE

Table 11
Amino acid sequences of alpha toxin Alpha toxin Staphylococcus aureus adsdiniktgttdigsnttvktgdlvtydkengmhkkvfysfiddknhnkkllvirtkgtiagqyrvyseeganksglawpsafkvqlqlpdnevaqisdyyprnsidtkeymstltygfngnvtgddtgkiggliganvsightlkyvqpdfktilesptdkkvgwkvifnnmvnqnwgpydrd swnpvygnqlfmktrngsmkaadnfldpnkassllssgfspdfatvitmdrkaskqqtnidviyervrddyqlhwtstnwkgtntkdkwtdrsserykidwekeemtn (SEQ ID NO: 39) Mutant alpha toxin H35L S. aureus adsdiniktgttdigsnttvktgdlvtydkengmlkkvfysfiddknhnkkllvirtkgtiagqyrvyseeganksglawpsafkvqlqlpdnevaqisdyyprnsidtkeymstltygfngnvtgddtgkiggliganvsightlkyvqpdfktilesptdkkvgwkvifnnmvnqnwgpydrds wnpvygnqlfmktrngsmkaadnfldpnkassllssgfspdfatvitmdrkaskqqtnidviyervrddyqlhwtstnwkgtntkdkwtdrsserykidwekeemtn (SEQ ID NO:40)

Table 12
Exemplary amino acid sequences of antibodies that specifically bind to S. aureus surface antigen IsdH Description VH or VL sequence (with CDRs in bold) CDR1 CDR2 CDR3 mAb 2F4 VH EVQLLESGGGL
VQPGGSLRLSCA
ASGFTFS PYMMQ
WVRQAPGKGLEW
VS SIWPSGGKTYY
ADSVKG RFTISRDN
SKNTLY
LQMNSLRAEDTAV
YYCAR VRRGGATD
Y WGQGTLVTVSS
(SEQ ID NO: 80) PYMMQ
(SEQ ID NO: 90) SIWPS
GGKT
YYAD
SVKG (SEQ ID NO: 91) VRRG
GATDY (SEQ ID NO: 92) mAb 2F4 VL DIQMTQSPATLSVSP
GERATLSC RASQSVS
SNLG WYQQKPGQAP
RLLIY GASTRAT GIPT
RFSGSGSGTEFTLTISSLQS
EDFATYYC QQYQNW
PLLT FGGGTKVEIK
(SEQ ID NO: 81) RASQ
SVSSN
LG
(SEQ ID NO: 93) GAST
RAT
(SEQ ID NO: 94) QQYQ
NWPL
LT
(SEQ ID NO: 95) mAb A7 VH EVQLLESGGGLVQPG
GSLRLSCAASGFTFS N
YYMW WVRQAPGKGL
EWVS VIGPSGGPTQY
ADSVKG RFTISRDNSK
NTLY
LQMNSLRAEDTAVYY
CAR WGGRYSVFET W
GQGTMVTVSS
(SEQ ID NO: 82) NYYMW
(SEQ ID
NO: 96) VIGPS
GGPTQ
YADS
VKG
(SEQ ID NO: 97) WGGR
YSVFET (SEQ ID
NO: 98) mAb A7 VL DIQMTQSPATLSVSPGG
RATLSC RASQSVRKN
VA WYQQKPGQPPRLL
IY GASTRAT GVPARFS
GSGSGTEFTLTISRMQP
EDFVVYHC QQYSSW
PAF GQGTMVEIN
(SEQ ID NO: 83) RASQ
SVRKN
V.A.
(SEQ ID NO: 99) GASTR
AT
(SEQ ID NO: 100) QQYS
SWPAF
(SEQ ID NO: 101) mAb 1C1 VH EVQLLESGGGLVQ
PGGSLRLSCAASGF
TFS RYFMG WVRQA
PGKGLEWVS SIYSS
GGYTSYADSVKG R
FTISRDNSKNTLY
LQMNSLRAEDTAVY
YCAR RWRDGTFDY
WGQGTLVTVSS
(SEQ ID NO: 84) RYFMG (SEQ ID NO: 102) SIYSSG
GYTS
YADS
VKG
(SEQ ID NO: 103) RWR
DGTFDY (SEQ ID NO: 104) mAb 1C1 VL DIQMTQSPSSLSASIG
DRVTISC RASQSVRE
YLN WYQQKPGKAPK
LLIF AASSLQS GVPSR
FSGSGSGTDFTLTISSLQP
EDFATYYC QQSYST
RFT FGPGTKVDIK
(SEQ ID NO: 85) RASQSVREYLN (SEQ ID NO: 105) A.A.S.S.
LQS
(SEQ ID NO: 106) QQSY
STRFT (SEQ ID NO: 107) IsdH0003 VH QVQLQQSGAEVKKPG
SSVKVSCKASGGTFS S
YPIS WVRQAPGQGLEW
MG KIIPIFGTTNYAQK
FQG RVTITADESTSTAY
MELSSLRSEDTAIYYCA
SPNRPYNIGWHYYFD
Y WGKGTLVTVSS
(SEQ ID NO: 86) SYPIS
(SEQ ID NO: 108) KIIPIF
GTTNY
AQKF
QG
(SEQ ID NO: 109) PNRP
YNIG
WHYY
FDY
(SEQ ID NO: 110) IsdH0003 VL QSVLTQPASVSGSPGQ
SITISC TGTSSDVGGY
NYVS WYQQHPGKAP
KLMIY EGSKRPS GVS
NRFSGSRSGNTASLTISGL
QAEDEADYYC SSYT
TRSTRV FGGGTKLTVL
(SEQ ID NO: 87) TGTSS
DVGG
YNYVS
(SEQ ID NO: 111) EGSK
R.P.S.
(SEQ ID NO: 112) SSYT
TRST
RV
(SEQ ID NO: 113) IsdH0016 VH EVQLLESGGGLVQPG
GSLRLSCAASGFTFS S
YAMS WVRQAPGKGL
EWVS AISGSGGSTYY
ADSVKG RFTISRDNS
KNTLY
LQMNSLRAEDTAVY
YCAR DQDEGRANN
WWIPPGGR WGQG
TMVTVSS
(SEQ ID NO: 88) SYAMS
(SEQ ID NO: 114) AISG
SGGST
YYAD
SVKG
(SEQ ID NO: 115) DQDE
GRAN
N.W.W.I.
PPGGR
(SEQ ID NO: 116) IsdH0016 VL SSELTQDPTLSVAL
GQTVRITC QGDSL
RRSFAS WYQKKPG
QAPVLLIY GQNKR
PA GIPDRFSGSRSGN
SASLTITGAQ
AEDEADYYC NSRD
ARLNPYIL FGGGTKLTVL
(SEQ ID NO: 89) QGDS
LRRS
F.A.S.
(SEQ ID NO: 117) GQNKRPA
(SEQ ID NO: 118) NSRD
ARLN
PYIL
(SEQ ID NO: 119)

Table 13
nucleotide sequences encoding the VH and VL amino acid sequences for mAbs directed against S. aureus surface antigens IsdH and ClfA SEQ ID NO: Description Subsequence SEQ ID NO: 120 Nucleotide sequence of VH mAb 2F4 GAAGTTCAATTGTTAGAGTCTGGTGGCGGTCTTGTTCAGCCTGGTGGTTCTTTACGTCTTTCTTGCGCTGCTTCCGGATTCACTTTCTCTCCTTACATGATGCAGTGGGTTCGCCAAGCTCCTGGTAAAGGTTTGGAGTGGGTTTCTTCTATCTGGCCTTCTGGTGGCAAGACTTATTATGCTGACTCCGTTAAAGGTCGCTTCACTATCTCTAGAGACAACTCTAAGAATACTCTCT ACTTGCAGATGAACAGCTTAAGGGCTGAGGACACGGCCGTGTATTACTGTGCGAGAGTGCGGAGGGGGGGAGCTACTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCAAGC SEQ ID NO: 121 Nucleotide sequence of VL mAb 2F4 GACATCCAGATGACCCAGTCTCCAGCCACCCTGTCTGTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGCAGCAACTTAGGCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATCCACCAGGGCCACTGGTATCCCAACCAGGTTCAGTGGCAGTGGGTCTGGGACAGGTTCACTCTCACCATCAGCAGCTGCAGTC TGAAGATTTTGCAACTTATTACTGTCAGCAGTATCAGAACTGGCCCTTGCTCACTTTCGGCGGAGGGACCAAGGTGGAAATCAAA SEQ ID NO: 122 Nucleotide sequence of VH mAb A7 GAAGTTCAATTGTTAGAGTCTGGTGGCGGTCTTGTTCAGCCTGGTGGTTCTTTACGTCTTTCTTGCGCTGCTTCCGGATTCACTTTCTCTAATTACTATATGTGGTGGGTTCGCCAAGCTCCTGGTAAAGGTTTGGAGTGGGTTTCTGTTATCGGTCCTTCTGGTGGCCCTACTCAGTATGCTGACTCCGTTAAAGGTCGCTTCACTATCTCTAGAGACAACTCTAAGAATACTCT CTACTTGCAGATGAACAGCTTAAGGGCTGAGGACACGGCCGTGTATTACTGTGCGAGATGGGGTGGGAGGTACTCTGTATTTGAAACCTGGGGCCAAGGGACAATGGTCACCGTCTCAAGC SEQ ID NO: 123 Nucleotide sequence of VL mAb A7 GACATCCAGATGACCCAGTCTCCAGCCACTCTGTCTGTGTCTCCAGGGGGAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGTGTTAGAAAAAACGTAGCCTGGTATCAGCAGAAACCTGGCCAGCCTCCCAGGCTCCTCATCTATGGTGCATCCACCAGGGCCACTGGTGTCCCAGCCAGTTCAGTGGCAGTGGGTCTGGGACAGAGTTCACTCTCACCATCAGCAGGATGCAGCCTGA AGATTTTGTAGTTTATCACTGTCAGCAGTATAGTAGCTGGCCGGCGTTCGGCCAGGGGACCATGGTGGAAATCAAC SEQ ID NO: 124 Nucleotide sequence of VH mAb 1C1 GAAGTTCAATTGTTAGAGTCTGGTGGCGGTCTTGTTCAGCCTGGTGGTTCTTTACGTCTTTCTTGCGCTGCTTCCGGATTCACTTTCTCTCGTTACTTTATGGGTTGGGTTCGCCAAGCTCCTGGTAAAGGTTTGGAGTGGGTTTCTTCTATCTATTCTTCTGGTGGCTATACTTCTTATGCTGACTCCGTTAAAGGTCGCTTCACTATCTCTAGAGACAACTCTAAGAATACTCTCT ACTTGCAGATGAACAGCTTAAGGGCTGAGGACACGGCCGTGTATTACTGTGCGAGACGGTGGCGAGATGGCACCTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCAAGC SEQ ID NO: 125 Nucleotide sequence of VL mAb 1C1 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTATTGGAGACAGAGTCACCATCTCTTGCCGGGCAAGTCAGAGCGTTAGAGAGTATCTAAATTGGTATCAACAAAACCAGGGAAAGCCCCTAAACTCCTGATCTTTGCTGCATCCAGTTTGCAGAGTGGGGTCCCATCAAGATTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAG ATTTTGCAACTTATTACTGTCAACAGAGTTACAGTACCCGATTCACTTTCGGCCCTGGGACCAAAGTGGACATCAAA SEQ ID NO: 126 Nucleotide sequence of VH mAb IsdH0003 CAGGTACAGCTGCAGCAGTCAGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCTTCTGGAGG
CACCTTCAGCAGCTATCCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAAAGATCATCCCTA
TCTTTGGTACAACAAACTACGCGCAGAAGTTCCAGGGCAGAGTCACGATTACCGCGGACGAATCCACGAGCACTGCCTAC
ATGGAACTGAGCAGCCTGAGATCTGAGGACACGGCCATATATTACTGTGCGAGCCCCAATCGACCCTATAACATTGGCTG
GCACTACTACTTTGACTACTGGGGCAAAGGAACCCTGGTCACCGTCTCCTCA SEQ ID NO: 127 Nucleotide sequence of VL mAb IsdH0003 CAGTCTGTGCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTTCCTGCACTGGAACCAGCAGTGACGTTGGTGGTTATAACTATGTCTCCTGGTACCAACAACACCCAGGCAAAGCCCCCAAACTCATGATTTATGAGGGCAGTAAGCGGCCCTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAGGTCTGGCAACACGGCCTCCCTGACAATCTCTGGGCTCCA GGCTGAGGACGAGGCTGATTATTACTGCAGCTCATATACAACCAGGAGCACTCGAGTCTTCGGCGGAGGGACCAAGCTGACCGTCCTA SEQ ID NO: 128 Nucleotide sequence of VH mAb IsdH0016 GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTG TATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCAAGAGATCAGGACGAAGGTAGAGCGAACAACTGGTGGATCCCCCCCGGGGGTCGCTGGGGCCAGGGGACAATGGTCACCGTCTCGAGT SEQ ID NO: 129 Nucleotide sequence of VL mAb IsdH0016 TCTTCTGAGCTGACTCAGGACCCTACTCTGTCTGTGGCCCTGGGACAGACAGTCAGAATCACATGCCAAGGACAGCCTCCGAAGATCTTTTGCAAGTTGGTACCAGAAGAAGCCAGGACAGGCCCCTGTACTTCTCATCTATGGTCAAAATAAGCGGCCCGCAGGGATCCCAGACCGATTCTCTGGCTCCAGGTCAGGAAACTCAGCTTCGTTGACCATCACAGGGGCTCAGGCGGAAGAT GAGGCTGACTATTACTGTAATTCCCGCGACGCCAGACTTAACCCTTATATACTCTTCGGCGGTGGGACCAAGCTGACCGTCCTA

Table 14
Exemplary amino acid sequences of antibodies that specifically bind to S. aureus surface antigen ClfA Description VH or VL sequence (with CDRs in bold) CDR1 CDR2 CDR3 mAb 23D6 VH QVQLVESGGGVV
QPGRSLRLSCAAS
VFTFS SYGMH WVR
QAPGKGLEWVA L
IWFDGSNEYYADS
VKG RFTISRDNSKNTLY
LQMNSLRAEDTAV
YYCARD RGGGYY
YYGMDV WGQGT
TVTVSS (SEQ ID NO:132) SYGMH
(SEQ ID NO:133) LIWFD
GSNEY
YADSV
KG
(SEQ ID NO: 134) RGGG
YYYY
GMDV
(SEQ ID
NO: 135) mAb 23D6 VL DIQMTQSPSSLSAS
VGDRVTITC RASQ
GIRNDLG WYQQK
PGKAPKRLIY AAS
SLQS GVPSRFSGSG
SGTEFTLTISSLQP
EDFATYYC LQHNS
YPYT FGQGTKLEIK (SEQ ID NO:136) RASQG
IRND
LG
(SEQ ID NO: 137) AASSL
QS
(SEQ ID NO: 138) LQHNS
YPYT
(SEQ ID NO: 139) mAb 27H4 VH QVQLVQSGAEVKK
PGASVKVSCKTSG
YTFT SYGIS WVRQA
PGQGHEWMG WISS
YNGNTNYAQKLQ
G RVTMTSDTSTSTA
YMELRSLRSDDTA
VYYCARI AARGYY
YGMD VWGQGTTVTVSS (SEQ ID NO:140) SYGIS
(SEQ ID NO: 141) WISSY
NGNT
NYAQK
LQG
(SEQ ID NO: 142) AARG
YYYG
M.D.
(SEQ ID NO: 143) mAb 27H4 VL EIVLTQSPGTLLSLS
PGERATLSC RASQ
SISGSYLA WYQQK
PGQAPRLLIY GAS
SRAT GIPDRFSGSG
SGTDFTLTISRLE
PEDFAVYYC QQY
GSSPWT FGQGTK
VEIK (SEQ ID NO:144) RASQ
SISGS
YLA
(SEQ ID NO: 145) GASS
RAT
(SEQ ID NO: 146) QQYSS
WPAF
(SEQ ID NO: 147)

[00241] The previous examples and tables are intended to be illustrative and not to limit the present disclosure in any way. Other embodiments of the disclosed devices and methods will become apparent to those skilled in the art upon consideration of the description and practical application of the devices and methods disclosed herein.

[00242] The following are embodiments of the present invention:

1. An isolated antibody or antigen-binding fragment thereof that specifically binds to the IsdH antigen of Staphylococcus aureus (S. aureus), which is a surface determinant,

wherein the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VH), each of which contains three complementarity determining regions (CDR1, CDR2 and CDR3),

and where:

a. CDR1 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 90, 96, 102, 108 or 114;

b. CDR2 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 91, 97, 103, 109 or 115; And

With. CDR3 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 92, 98, 104, 110 or 116;

and/or

d. CDR1 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 93, 99, 105, 111 or 117;

e. CDR2 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 94, 100, 106, 112 or 118; And

f. CDR3 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 95, 101, 107, 113 or 119.

2. The isolated antibody or antigen-binding fragment thereof according to embodiment 1, wherein the isolated antibody or antigen-binding fragment thereof comprises:

a. CDR1 VH identical to or containing 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 90, 96, 102, 108 or 114;

b. CDR2 VH identical to or containing 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 91, 97, 103, 109 or 115;

With. CDR3 VH identical to or containing 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 92, 98, 104, 110 or 116;

d. CDR1 VL identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 93, 99, 105, 111 or 117;

e. CDR2 VL identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 94, 100, 106, 112 or 118; And

f. CDR3 VL identical to or containing 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 95, 101, 107, 113 or 119.

3. The isolated antibody or antigen binding fragment thereof of embodiment 1 or 2, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL correspond to a set of amino acid sequences selected from the group consisting of SEQ ID NO: 90, 91, 92, 93, 94 and 95; SEQ ID NO: 96, 97, 98, 99, 100 and 101; SEQ ID NO: 102, 103, 104, 105, 106 and 107; SEQ ID NOs: 108, 109, 110, 111, 112 and 113 and SEQ ID NOs: 114, 115, 116, 117, 118 and 119.

4. An isolated antibody or antigen-binding fragment thereof that specifically binds to the surface determinant IsdH antigen of Staphylococcus aureus (S. aureus), wherein the VH amino acid sequence is at least 80%, 85%, 90%, 95% or 100% identical amino acid sequence SEQ ID NO: 80, 82, 84, 86 or 88.

5. The isolated antibody or antigen binding fragment thereof of embodiment 4, wherein the amino acid sequence of VH is identical to or contains 1-10 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 80, 82, 84, 86 or 88.

6. An isolated antibody or antigen-binding fragment thereof that specifically binds to the surface determinant antigen IsdH of Staphylococcus aureus (S. aureus), wherein the VL amino acid sequence is at least 80%, 85%, 90%, 95% or 100% identical amino acid sequence SEQ ID NO: 81, 83, 85, 87 or 89.

7. The isolated antibody or antigen binding fragment thereof of embodiment 6, wherein the amino acid sequence of VL is identical to or contains 1-10 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 81, 83, 85, 87 or 89.

8. The isolated antibody or antigen binding fragment thereof according to any one of embodiments 1-7, wherein the amino acid sequence is at least 80%, 85% VH. 90%, 95% or 100% identical to the amino acid sequence of SEQ ID NO: 80, 82, 84, 86 or 88, and the amino acid sequence of VL is at least 80%, 85%. 90%, 95% or 100% identical to the amino acid sequence of SEQ ID NO: 81, 83, 85, 87 or 89.

9. The isolated antibody or antigen binding fragment thereof of Embodiment 8, wherein the amino acid sequence VH corresponds to the amino acid sequence of SEQ ID NO: 80, 82, 84, 86 or 88, and the amino acid sequence VL corresponds to the amino acid sequence of SEQ ID NO: 81, 83, 85 , 87 or 89.

10. The isolated antibody or antigen binding fragment thereof according to any one of embodiments 1-8, wherein VH and VL are selected from the group consisting of SEQ ID NOs: 80 and 81; SEQ ID NO: 82 and 83; SEQ ID NO: 84 and 85; SEQ ID NO: 86 and 87 and SEQ ID NO: 88 and 89.

11. The isolated antibody or fragment of any one of embodiments 1-3, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL correspond to a set of amino acid sequences selected from the group consisting of SEQ ID NO: 90, 91, 92, 93, 94 and 95.

12. The isolated antibody or antigen binding fragment thereof of Embodiment 1 or 2, wherein the VH amino acid sequence comprises four VH framework regions (FR1, FR2, FR3, FR4), and wherein the four VH framework regions have amino acid sequences that are at least approximately 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four VH framework regions in SEQ ID NO: 80, 82, 84, 86 or 88.

13. The isolated antibody or antigen binding fragment thereof of Embodiment 1 or 2, wherein the VL amino acid sequence comprises four VL framework regions (FR1, FR2, FR3, FR4), and wherein the four VL framework regions have amino acid sequences that are at least approximately 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four VL framework regions in SEQ ID NO: 81, 83, 85, 87 or 89.

14. The isolated antibody or antigen binding fragment thereof of Embodiment 1 or 2, wherein the four VH framework regions have amino acid sequences that are at least about 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four frameworks VH regions of SEQ ID NO: 80, 82, 84, 86 or 88, and wherein the four VL framework regions have amino acid sequences that are at least about 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acids sequences of the four VL framework regions in SEQ ID NO: 81, 83, 85, 87 or 89.

15. The isolated antibody or antigen binding fragment thereof of any one of embodiments 1-14, wherein VH and VL correspond to SEQ ID NOs 80 and 81.

16. The isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-15, wherein the antibody or antigen-binding fragment thereof is characterized by at least one of the following:

a. a dissociation constant (KD) for S. aureus surface antigen of approximately 70 nM or less;

b. the ability to reduce the ability of S. aureus to evade opsonophagocytosis by at least 50%, as measured by an opsonophagocytic killing assay; or

c. ability to reduce the number of colony forming units (CFU) of S. aureus by at least 50%, as measured using a bacteremia model.

17. An isolated antibody or antigen-binding fragment thereof that specifically binds to the surface determinant IsdH antigen of Staphylococcus aureus (S. aureus), and wherein the antibody or antigen-binding fragment thereof is characterized by at least one of the following:

a. a dissociation constant (KD) for S. aureus surface antigen of approximately 70 nM or less;

b. the ability to reduce the ability of S. aureus to evade opsonophagocytosis by at least 50%, as measured by an opsonophagocytic killing assay;

c. the ability to reduce the number of S. aureus colony forming units (CFU) by at least 50%, as measured using a bacteremia model;

d. ability to reduce immune cell infiltration, bacterial load, and proinflammatory cytokine release as measured in an animal model of organ loading.

18. The isolated antibody or antigen binding fragment thereof of embodiment 17, having the amino acid sequence of the antibody or antigen binding fragment thereof of any one of embodiments 1-16.

19. A composition comprising an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18 and a pharmaceutically acceptable excipient.

20. An isolated nucleic acid encoding the amino acid sequence of any one of embodiments 1-15.

21. An isolated antibody or antigen-binding fragment thereof that specifically binds to the ClfA antigen of Staphylococcus aureus (S. aureus), which is a surface determinant,

wherein the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VH), each of which contains three complementarity determining regions (CDR1, CDR2 and CDR3),

and where:

a. CDR1 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 133 or 141;

b. CDR2 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 134 or 142;

And

With. CDR3 VH is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 135 or 143;

and/or

d. CDR1 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 137 or 145;

e. CDR2 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 138 or 146;

And

f. CDR3 VL is identical to or contains 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 139 or 147.

22. The isolated antibody or antigen-binding fragment thereof of embodiment 21, wherein the isolated antibody or antigen-binding fragment thereof comprises:

a. CDR1 VH identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 133 or 141;

b. CDR2 VH identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 134 or 142;

With. CDR3 VH identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 135 or 143;

d. CDR1 VL identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 137 or 145;

e. CDR2 VL identical to or containing 1, 2 or 3 mutations of amino acid residues relative to the amino acid sequence of SEQ ID NO: 138 or 146;

And

f. CDR3 VL identical to or containing 1, 2 or 3 amino acid residue mutations relative to the amino acid sequence SEQ ID NO: 139 or 147.

23. The isolated antibody or antigen binding fragment thereof of embodiment 21 or 22, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL correspond to a set of amino acid sequences selected from the group consisting of SEQ ID NO: 133, 134, 135, 137, 138 and 139 and SEQ ID NO: 137, 138, 139, 145, 146 and 147.

24. An isolated antibody or antigen-binding fragment thereof that specifically binds to the surface determinant antigen ClfA of Staphylococcus aureus (S. aureus), wherein the VH amino acid sequence is at least 80%, 85%, 90%, 95% or 100% identical amino acid sequence SEQ ID NO: 132 or 140.

25. The isolated antibody or antigen binding fragment thereof of embodiment 24, wherein the amino acid sequence of VH is identical to or contains 1-10 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 132 or 140.

26. An isolated antibody or antigen-binding fragment thereof that specifically binds to the surface determinant antigen ClfA of Staphylococcus aureus (S. aureus), where the amino acid sequence VL is at least 80%, 85%, 90%, 95% or 100% identical amino acid sequence SEQ ID NO: 136 or 144.

27. The isolated antibody or antigen binding fragment thereof of embodiment 26, wherein the amino acid sequence of VL is identical to or contains 1-10 amino acid residue mutations relative to the amino acid sequence of SEQ ID NO: 136 or 144.

28. The isolated antibody or antigen binding fragment thereof of any one of embodiments 21-27, wherein the amino acid sequence of VH is at least 80%, 85%, 90%, 95% or 100% identical to the amino acid sequence of SEQ ID NO: 132 or 140, and the amino acid sequence of VL is at least 80%, 85%, 90%, 95% or 100% identical to the amino acid sequence of SEQ ID NO: 136 or 144.

29. The isolated antibody or antigen binding fragment thereof of embodiment 28, wherein the amino acid sequence VH corresponds to the amino acid sequence of SEQ ID NO: 132 or 140, and the amino acid sequence VL corresponds to the amino acid sequence of SEQ ID NO: 136 or 144.

30. The isolated antibody or antigen binding fragment thereof of any one of embodiments 21-28, wherein VH and VL are selected from the group consisting of SEQ ID NOs: 132 and 136 and SEQ ID NOs: 140 and 144.

31. The isolated antibody or fragment of any one of embodiments 21-23, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL correspond to a set of amino acid sequences selected from the group consisting of SEQ ID NO: 141, 142, 143, 144, 145, 146 and 147.

32. The isolated antibody or antigen binding fragment thereof of Embodiment 21 or 22, wherein the VH amino acid sequence comprises four VH framework regions (FR1, FR2, FR3, FR4), and wherein the four VH framework regions have amino acid sequences that are at least approximately 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four VH framework regions in SEQ ID NO: 132 or 140.

33. The isolated antibody or antigen binding fragment thereof of Embodiment 21 or 22, wherein the VL amino acid sequence comprises four VL framework regions (FR1, FR2, FR3, FR4), and wherein the four VL framework regions have amino acid sequences that are at least approximately 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four VL framework regions in SEQ ID NO: 136 or 144.

34. The isolated antibody or antigen binding fragment thereof of Embodiment 21 or 22, wherein the four VH framework regions have amino acid sequences that are at least about 80%, 85%, 90%, 95%, or 100% identical to the corresponding amino acid sequences of the four frameworks. the VH regions of SEQ ID NO: 132 or 144, and wherein the four VL framework regions have amino acid sequences that are at least about 80%, 85%, 90%, 95% or 100% identical to the corresponding amino acid sequences of the four VL framework regions in SEQ ID NO: 136 or 144.

35. The isolated antibody or antigen binding fragment thereof of any one of embodiments 21-34, wherein VH and VL correspond to SEQ ID NOs 140 and 144.

36. A composition comprising an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 21-35 and a pharmaceutically acceptable excipient.

37. An isolated nucleic acid encoding the amino acid sequence of any one of embodiments 21-35.

38. A composition comprising an isolated antibody or an antigen-binding fragment thereof that specifically binds to S. aureus alpha-toxin (AT), and an isolated antibody or an antigen-binding fragment thereof that specifically binds to an S. aureus surface determinant antigen.

39. The composition of embodiment 38, wherein the S. aureus surface determinant antigen is selected from the group consisting of SdrC, SdrD, SdrE, ClfA, ClfB, IsdA, IsdB, IsdC, IsdE, IsdH, SpA, FnbA and PNAG.

40. The composition of embodiment 38 or 39, wherein the surface determinant antigen is IsdH.

41. The composition of embodiment 40, wherein the isolated antibody or antigen binding fragment thereof that specifically binds IsdH is the antibody or antigen binding fragment thereof of any one of embodiments 1-18.

42. The composition of embodiment 38 or 39, wherein the surface determinant antigen is ClfA.

43. The composition of embodiment 42, wherein the isolated antibody or antigen binding fragment thereof that specifically binds ClfA is the antibody or antigen binding fragment thereof of any one of embodiments 21-35.

44. The composition according to any of embodiments 38-43, where the isolated antibody or antigen-binding fragment thereof, which specifically binds AT, contains:

a. CDR1 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO: 7, 10, 13 or 69;

b. CDR2 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NO 8, 11, 14, 17, 70 or 75; And

c. CDR3 VH containing an amino acid sequence identical to or containing 1, 2 or 3 mutations of amino acid residues with respect to SEQ ID NOs 9, 12, 15, 16, 18, 65, 66, 67, 71, 72, 76 or 78;

and/or

d. CDR1 VL containing the amino acid sequence of SEQ ID NO: 1 or 4;

e. CDR2 VL containing the amino acid sequence of SEQ ID NO: 2, 5, 73 or 77; And

f. CDR3 VL containing the amino acid sequence SEQ ID NO: 3, 6, 64, 68 or 74.

45. The composition according to any of embodiments 38-43, where the isolated antibody or antigen-binding fragment thereof, which specifically binds AT, contains:

a. CDR1 VH containing the amino acid sequence of SEQ ID NO: 7, 10, 13 or 69;

b. CDR2 VH containing the amino acid sequence SEQ ID NO: 8, 11, 14, 17, 70 or 75;

c. CDR3 VH containing the amino acid sequence SEQ ID NO: 9, 12, 15, 18, 16, 65, 66, 67, 71, 72, 76 or 78;

d. CDR1 VL containing the amino acid sequence of SEQ ID NO: 1 or 4;

e. CDR2 VL containing the amino acid sequence of SEQ ID NO: 2, 5, 73 or 77; And

f. CDR3 VL containing the amino acid sequence SEQ ID NO: 3, 6, 64, 68 or 74.

46. The composition according to any one of embodiments 38-45, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL of the isolated antibody or antigen binding fragment thereof that specifically bind AT correspond to the amino acid sequences of SEQ ID NO: 7 , 8, 9, 1, 2 and 3; SEQ ID NO: 10, 11, 12, 1, 2 and 3; SEQ ID NO: 13, 14, 15, 4, 5 and 6; SEQ ID NO: 7, 17, 18, 1, 2 and 3; SEQ ID NO: 7, 8, 16, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 64; SEQ ID NO; 7, 8, 66, 1, 2 and 64; SEQ ID NO: 7, 8, 67, 1, 2 and 68; SEQ ID NO: 7, 8, 67, 1, 2 and 64; SEQ ID NO: 7, 8, 78, 1, 2 and 64; SEQ ID NO: 7, 8, 65, 1, 2 and 68; SEQ ID NO: 69, 70, 71, 1, 2 and 68; SEQ ID NO: 7, 8, 72, 1, 73 and 74; SEQ ID NO: 69, 75, 71, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 2 and 68; SEQ ID NO: 69, 75, 76, 1, 77 and 74; SEQ ID NO: 69, 70, 71, 1, 77 and 74.

47. The composition of any one of embodiments 38-43, wherein the isolated antibody or antigen binding fragment thereof that specifically binds AT contains a heavy chain variable domain that is at least 90% identical to the amino acid sequence of SEQ ID NO: 20, 22, 24 , 26, 28, 41, 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61 or 62; and/or (iii) contain a light chain variable domain that is at least 90% identical to the amino acid sequence of SEQ ID NO: 19, 21, 23, 25, 27, 42, 44, 46, 48, 50, 52, 54, 56 , 58, 60 or 63.

48. The composition of any one of embodiments 38-43, wherein the isolated antibody or antigen binding fragment thereof that specifically binds AT contains a heavy chain variable domain that has the amino acid sequence of SEQ ID NO: 20, 22, 24, 26, 28, 41 , 43, 45, 47, 49, 51, 53, 55, 57, 79, 59, 61, or 62, and (iii) contains a light chain variable domain having the amino acid sequence SEQ ID NO: 19, 21, 23, 25 , 27, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 or 63.

49. The composition according to any one of embodiments 38-48, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL of the isolated antibody or antigen binding fragment thereof that specifically bind AT correspond to the amino acid sequences of SEQ ID NO: 69 , 70, 71, 1, 2 and 68.

50. The composition according to any one of embodiments 38-49, wherein the isolated antibody or antigen binding fragment thereof that specifically binds AT contains a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 57, and contains a light chain variable domain, wherein the domain has the amino acid sequence SEQ ID NO: 58.

51. The composition of any one of embodiments 38-50, wherein the isolated antibody or antigen binding fragment thereof that specifically binds AT further comprises a variant Fc region.

52. The composition of embodiment 51, wherein the isolated antibody or antigen binding fragment thereof that specifically binds AT comprises: 130 and SEQ ID NO: 131.

53. The composition of any one of embodiments 30 or 40-41, further comprising an isolated antibody or antigen-binding fragment thereof that specifically binds ClfA.

54. The composition of embodiment 53, wherein the isolated antibody or antigen binding fragment thereof that specifically binds ClfA is the antibody or antigen binding fragment thereof of any one of embodiments 21-35.

55. A method for determining the presence of S. aureus in a test sample, comprising contacting the test sample with an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-15 or 21-34 and a detectable label.

56. The method of embodiment 55, comprising:

a. contacting the test sample with an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof binds to an S. aureus surface determinant antigen to form a first complex;

b. bringing the complex into contact with a detectable label, wherein the detectable label binds to the antibody or an antigen-binding fragment thereof or to an antigen to form a second complex; And

c. detecting the presence of S. aureus in the test sample based on the signal created by the detectable label in the second complex, wherein the presence of S. aureus is directly correlated with the signal created by the detectable label.

57. The method of embodiment 55, comprising:

a. contacting the test sample with an antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof binds to an S. aureus surface determinant antigen to form a first complex;

b. bringing the complex into contact with a detectable label, wherein the detectable label competes with the antigen for binding to the antibody or antigen-binding fragment thereof so as to form a second complex; And

c. detecting the presence of S. aureus in the test sample based on the signal created by the detectable label in the second complex, wherein the presence of S. aureus indirectly correlates with the signal created by the detectable label.

58. The method as in any one of embodiments 55-57, wherein the sample is a patient sample, and the method further includes diagnosing a patient with an S. aureus infection.

59. The method as in any one of embodiments 55-58, wherein the method is adapted for use in an automatic system or semi-automatic system.

60. A method for determining the presence of S. aureus in a test sample, comprising contacting the test sample with the composition of claim 19 or 36 and at least one detectable label.

61. The method of embodiment 60, comprising:

a. contacting the test sample with the composition of embodiment 13, wherein the composition binds to the S. aureus surface determinant antigen and the S. aureus secreted toxin polypeptide;

b. bringing the test sample into contact with at least one detectable label, wherein the at least one detectable label binds to a surface determinant antigen, a secreted toxin, or an antibody, or an antigen-binding fragment thereof, that is associated with a surface determinant antigen or secreted toxin ; And

c. detecting the presence of S. aureus in the test sample based on a signal created by the at least one detectable label, wherein the presence of S. aureus is directly correlated with the signal created by the at least one detectable label.

62. The method of embodiment 60, comprising:

a. contacting the test sample with the composition of embodiment 13, wherein the composition binds to the S. aureus surface determinant antigen and the S. aureus secreted toxin polypeptide;

b. contacting the test sample with at least one detectable label, wherein the at least one detectable label competes with a surface determinant antigen or secreted toxin for binding to the antibody in the composition of embodiment 13; And

c. detecting the presence of S. aureus in the test sample based on a signal created by at least one detectable label, wherein the presence of S. aureus indirectly correlates with the signal created by at least one detectable label.

63. The method as in any one of embodiments 60-62, wherein the sample is a patient sample, and the method further includes diagnosing a patient with an S. aureus infection.

64. The method as in any one of embodiments 60-63, wherein the method is adapted for use in an automatic system or semi-automatic system.

65. A method of treating an S. aureus infection in a patient, comprising administering to the patient an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18 or 21-35 or a composition according to any one of embodiments 19, 36 or 38-54.

66. A method of preventing or reducing the severity of S. aureus-associated sepsis in a patient, comprising administering to the patient an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18 or 21-35, or a composition according to any one of embodiments 19, 36 or 38-54.

67. A method of delaying the onset of sepsis associated with S. aureus infection in a patient, comprising administering to the patient an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18 or 21-35 or a composition according to any one of embodiments 19, 36 or 38 -54.

68. A method of preventing the onset of sepsis associated with S. aureus infection in a patient, comprising administering to the patient an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18 or 21-35 or a composition according to any one of embodiments 19, 36 or 38 -54.

69. A method of reducing the bacterial load of S. aureus in the bloodstream or heart of a patient, comprising administering to said patient an effective amount of an isolated antibody or antigen-binding fragment thereof according to any one of embodiments 1-18.

70. A method of reducing agglutination of S. aureus bacteria and/or the formation of thromboembolic lesions in a patient, comprising administering to said patient an effective amount of an isolated antibody or an antigen-binding fragment thereof according to any one of embodiments 1-18.

71. The method of any one of embodiments 65-70, further comprising administering an antibiotic to the patient.

72. The method of embodiment 69, wherein the antibiotic is penicillin, oxacillin, flucloxacillin, or vancomycin.

LIST OF SEQUENCES

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<110> MEDIMMUNE, LLC

<120> ANTIBODIES TO SURFACE DETERMINANTS S. AUREUS

<130>ATOX-150WO1

<140> PCT/US2013/068624

<141> 2013-11-06

<150> 61/782,405

<151> 2013-03-14

<150> 61/723.137

<151> 2012-11-06

<160> 148

<170> PatentIn version 3.5

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Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

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Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

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Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

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Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

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35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

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Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

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Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Trp Thr

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Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

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Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr

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Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

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Ser Val Ile Gly Thr Asp Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Ile Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

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Glu Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

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Arg Asp Arg Tyr Ser Ser Ser Asn His Tyr Asn Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

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Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

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Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 24

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 24

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Gly Thr Asp Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Ile Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Glu Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Ser Ser Asn His Tyr Asn Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 25

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 25

Ala Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln Asp Tyr Asn Tyr Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 26

<211> 125

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 26

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Asp Gly Arg Gln Val Glu Asp Tyr Tyr Tyr Tyr Tyr Gly Met

100 105 110

Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120 125

<210> 27

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 27

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 28

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 28

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Asp Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Val

85 90 95

Arg Asp Arg Tyr Ser Gly Asn Phe His Tyr Asn Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 29

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 29

gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctataag gcgtctagtt tagaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttatta ctgccaacag tataatagtt attggacgtt cggccaaggg 300

accaaggtgg aaatcaaa 318

<210> 30

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 30

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt agctacgaca tgcactgggt ccgccaagct 120

acaggaaaag gtctggagtg ggtctcaggt attggcactg ctggtgacac atattatcca 180

ggctccgtga agggccgatt caccatctcc agagaaaatg ccaagaactc cttgtatctt 240

caattgaaca gcctgagagc cggggacacg gctgtgtact tctgtgcaag agacaattat 300

agcagcaccg gggggtacta cggtatggac gtctggggcc aagggaccac ggtcaccgtc 360

tcctca 366

<210> 31

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 31

gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaaactcct gatctataag gcgtctagtt tagaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttatta ctgccaacag tataatagtt attggacgtt cggccaaggg 300

accaaggtgg aaatcaaa 318

<210> 32

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 32

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt aggtacgaca tgcactgggt ccgccaagct 120

acaggaaaag gtctggagtg ggtctcagtt attggtactg atggtgacac atactatcca 180

ggctccgtga agggccgatt catcatctcc agagaaaatg ccaagaactc cttgtatctt 240

gaaatgaaca gcctgagagc cggggacacg gctgtgtatt actgtgcaag agatcggtat 300

agcagctcga accactacaa cggtatggac gtctggggcc aagggaccac ggtcaccgtc 360

tcctca 366

<210> 33

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 33

gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaaggtcct gatctataag gcgtctagtt tagaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttatta ctgccaacag tataatagtt attggacgtt cggccaaggg 300

accaaggtgg aaatcaaa 318

<210> 34

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 34

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt caccttcagt aggtacgaca tgcactgggt ccgccaagct 120

acaggaaaag gtctggagtg ggtctcagtt attggtactg atggtgacac atactatcca 180

ggctccgtga agggccgatt catcatctcc agagaaaatg ccaagaactc cttgtatctt 240

gaaatgaaca gcctgagagc cggggacacg gctgtgtatt actgtgcaag agatcggtat 300

agcagctcga accactacaa cggtatggac gtctggggcc aagggaccac ggtcaccgtc 360

tcctca 366

<210> 35

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 35

gccatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctatgat gcatccagtt tacaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tggcacagat ttcactctca ccatcagcag cctgcagcct 240

gaagattttg caacttatta ctgtctacaa gattacaatt acccgtggac gttcggccaa 300

gggaccaagg tggaaatcaa a 321

<210> 36

<211> 375

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 36

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgacctgggt ccgccaggct 120

ccagggaagg ggctggaatg ggtctcagtt attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccgtc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatggg 300

aggcaggtcg aggattacta ctactactac ggtatggacg tctggggcca agggaccacg 360

gtcaccgtct cctca 375

<210> 37

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 37

gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60

atcacttgcc gggccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120

gggaaagccc ctaagctcct gatctataag gcgtctagtt tagaaagtgg ggtcccatca 180

aggttcagcg gcagtggatc tgggacagaa ttcactctca ccatcagcag cctgcagcct 240

gatgattttg caacttatta ctgccaacag tataatagtt attggacgtt cggccaaggg 300

accaaggtgg aaatcaaa 318

<210> 38

<211> 366

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 38

gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtacag cctctggatt caccttcagt agttacgaca tgcactgggt ccgccaagct 120

acaggaaaag gtctggagtg ggtctcagtt attgatactg ctggtgacac atactatcca 180

ggctccgtga agggccgatt caccatctcc agagaaaatg ccaagaactc cttgtatctt 240

caaatgaaca gcctgagagc cggggacacg gctgtgtatt actgtgtaag agataggtat 300

agtgggaact tccactacaa cggtatggac gtctggggcc aagggaccac ggtcaccgtc 360

tcctca 366

<210> 39

<211> 293

<212> PROTEIN

<213> Staphylococcus aureus

<400> 39

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser

1 5 10 15

Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn

20 25 30

Gly Met His Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn His

35 40 45

Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln

50 55 60

Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp

65 70 75 80

Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala

85 90 95

Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr

100 105 110

Met Ser Thr Leu Thr Tyr Gly Phe Asn Gly Asn Val Thr Gly Asp Asp

115 120 125

Thr Gly Lys Ile Gly Gly Leu Ile Gly Ala Asn Val Ser Ile Gly His

130 135 140

Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys Thr Ile Leu Glu Ser Pro

145 150 155 160

Thr Asp Lys Lys Val Gly Trp Lys Val Ile Phe Asn Asn Met Val Asn

165 170 175

Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser Trp Asn Pro Val Tyr Gly

180 185 190

Asn Gln Leu Phe Met Lys Thr Arg Asn Gly Ser Met Lys Ala Ala Asp

195 200 205

Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser Leu Leu Ser Ser Gly Phe

210 215 220

Ser Pro Asp Phe Ala Thr Val Ile Thr Met Asp Arg Lys Ala Ser Lys

225 230 235 240

Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu Arg Val Arg Asp Asp Tyr

245 250 255

Gln Leu His Trp Thr Ser Thr Asn Trp Lys Gly Thr Asn Thr Lys Asp

260 265 270

Lys Trp Thr Asp Arg Ser Ser Glu Arg Tyr Lys Ile Asp Trp Glu Lys

275 280 285

Glu Glu Met Thr Asn

290

<210> 40

<211> 293

<212> PROTEIN

<213> Staphylococcus aureus

<400> 40

Ala Asp Ser Asp Ile Asn Ile Lys Thr Gly Thr Thr Asp Ile Gly Ser

1 5 10 15

Asn Thr Thr Val Lys Thr Gly Asp Leu Val Thr Tyr Asp Lys Glu Asn

20 25 30

Gly Met Leu Lys Lys Val Phe Tyr Ser Phe Ile Asp Asp Lys Asn His

35 40 45

Asn Lys Lys Leu Leu Val Ile Arg Thr Lys Gly Thr Ile Ala Gly Gln

50 55 60

Tyr Arg Val Tyr Ser Glu Glu Gly Ala Asn Lys Ser Gly Leu Ala Trp

65 70 75 80

Pro Ser Ala Phe Lys Val Gln Leu Gln Leu Pro Asp Asn Glu Val Ala

85 90 95

Gln Ile Ser Asp Tyr Tyr Pro Arg Asn Ser Ile Asp Thr Lys Glu Tyr

100 105 110

Met Ser Thr Leu Thr Tyr Gly Phe Asn Gly Asn Val Thr Gly Asp Asp

115 120 125

Thr Gly Lys Ile Gly Gly Leu Ile Gly Ala Asn Val Ser Ile Gly His

130 135 140

Thr Leu Lys Tyr Val Gln Pro Asp Phe Lys Thr Ile Leu Glu Ser Pro

145 150 155 160

Thr Asp Lys Lys Val Gly Trp Lys Val Ile Phe Asn Asn Met Val Asn

165 170 175

Gln Asn Trp Gly Pro Tyr Asp Arg Asp Ser Trp Asn Pro Val Tyr Gly

180 185 190

Asn Gln Leu Phe Met Lys Thr Arg Asn Gly Ser Met Lys Ala Ala Asp

195 200 205

Asn Phe Leu Asp Pro Asn Lys Ala Ser Ser Leu Leu Ser Ser Gly Phe

210 215 220

Ser Pro Asp Phe Ala Thr Val Ile Thr Met Asp Arg Lys Ala Ser Lys

225 230 235 240

Gln Gln Thr Asn Ile Asp Val Ile Tyr Glu Arg Val Arg Asp Asp Tyr

245 250 255

Gln Leu His Trp Thr Ser Thr Asn Trp Lys Gly Thr Asn Thr Lys Asp

260 265 270

Lys Trp Thr Asp Arg Ser Ser Glu Arg Tyr Lys Ile Asp Trp Glu Lys

275 280 285

Glu Glu Met Thr Asn

290

<210> 41

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 41

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Met Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 42

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 42

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 43

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 43

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 44

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 44

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 45

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 45

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Arg Thr Gly His Tyr Met Gly Met Ser Leu Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 46

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 46

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 47

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 47

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Asn Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 48

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 48

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Tyr Ala Asp Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 49

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 49

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Asn Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 50

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 50

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 51

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 51

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Met Gly Met Ser Leu Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 52

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 52

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp Thr Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 53

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 53

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 54

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 54

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Tyr Ala Asp Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 55

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 55

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Asn Tyr Ser Ser Thr Gly Gly Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 56

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 56

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 57

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 57

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 58

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 58

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Tyr Ala Asp Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 59

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 59

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Asn Tyr Ser Pro Thr Gly Gly Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 60

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 60

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Lys Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Glu Ser Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 61

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 61

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Arg Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 62

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 62

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Arg Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Lys Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 63

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 63

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Val Lys Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Glu Ser Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 64

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 64

Gln Gln Tyr Asp Thr Tyr Trp Thr

15

<210> 65

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 65

Asp Arg Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val

1 5 10

<210> 66

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 66

Asp Arg Tyr Ser Arg Thr Gly His Tyr Met Gly Met Ser Leu

1 5 10

<210> 67

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 67

Asp Asn Tyr Ser Arg Thr Gly His Tyr Met Gly Met Asp Val

1 5 10

<210> 68

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 68

Lys Gln Tyr Ala Asp Tyr Trp Thr

15

<210> 69

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 69

Ser His Asp Met His

15

<210> 70

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 70

Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys Gly

1 5 10 15

<210> 71

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 71

Asp Arg Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val

1 5 10

<210> 72

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 72

Asp Asn Tyr Ser Pro Thr Gly Gly Tyr Tyr Gly Met Asp Val

1 5 10

<210> 73

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 73

Lys Ala Ser Ser Leu Lys Ser

15

<210> 74

<211> 8

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 74

Gln Gln Tyr Glu Ser Tyr Trp Thr

15

<210> 75

<211> 16

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 75

Gly Ile Gly Thr Arg Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys Gly

1 5 10 15

<210> 76

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 76

Asp Lys Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val

1 5 10

<210> 77

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 77

Lys Ala Ser Ser Leu Val Lys

15

<210> 78

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 78

Asp Arg Tyr Ser Pro Thr Gly His Tyr Met Gly Met Ser Leu

1 5 10

<210> 79

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 79

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 80

<211> 118

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 80

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Pro Tyr

20 25 30

Met Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Trp Pro Ser Gly Gly Lys Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Val Arg Arg Gly Gly Ala Thr Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 81

<211> 108

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 81

Asp Ile Gln Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Thr Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gln Asn Trp Pro Leu

85 90 95

Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 82

<211> 119

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 82

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr

20 25 30

Tyr Met Trp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Val Ile Gly Pro Ser Gly Gly Pro Thr Gln Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Trp Gly Gly Arg Tyr Ser Val Phe Glu Thr Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 83

<211> 106

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 83

Asp Ile Gln Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Gly Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Lys Asn

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro Arg Leu Leu Ile

35 40 45

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Val Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Arg Met Gln Pro

65 70 75 80

Glu Asp Phe Val Val Tyr His Cys Gln Gln Tyr Ser Ser Trp Pro Ala

85 90 95

Phe Gly Gln Gly Thr Met Val Glu Ile Asn

100 105

<210> 84

<211> 118

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 84

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Tyr

20 25 30

Phe Met Gly Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Tyr Ser Ser Gly Gly Tyr Thr Ser Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Trp Arg Asp Gly Thr Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 85

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 85

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Ile Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Ser Val Arg Glu Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Phe Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Arg Phe

85 90 95

Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys

100 105

<210> 86

<211> 124

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 86

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Pro Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Lys Ile Ile Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Ile Tyr Tyr Cys

85 90 95

Ala Ser Pro Asn Arg Pro Tyr Asn Ile Gly Trp His Tyr Tyr Phe Asp

100 105 110

Tyr Trp Gly Lys Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 87

<211> 110

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 87

Gln Ser Val Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Glu Gly Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Arg Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Thr Arg

85 90 95

Ser Thr Arg Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

<210> 88

<211> 126

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 88

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Gln Asp Glu Gly Arg Ala Asn Asn Trp Trp Ile Pro Pro

100 105 110

Gly Gly Arg Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120 125

<210> 89

<211> 109

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 89

Ser Ser Glu Leu Thr Gln Asp Pro Thr Leu Ser Val Ala Leu Gly Gln

1 5 10 15

Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Arg Ser Phe Ala

20 25 30

Ser Trp Tyr Gln Lys Lys Pro Gly Gln Ala Pro Val Leu Leu Ile Tyr

35 40 45

Gly Gln Asn Lys Arg Pro Ala Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

Arg Ser Gly Asn Ser Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ala Arg Leu Asn Pro

85 90 95

Tyr Ile Leu Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 90

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 90

Pro Tyr Met Met Gln

15

<210> 91

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 91

Ser Ile Trp Pro Ser Gly Gly Lys Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 92

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 92

Val Arg Arg Gly Gly Ala Thr Asp Tyr

15

<210> 93

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 93

Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Gly

1 5 10

<210> 94

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 94

Gly Ala Ser Thr Arg Ala Thr

15

<210> 95

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 95

Gln Gln Tyr Gln Asn Trp Pro Leu Leu Thr

1 5 10

<210> 96

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 96

Asn Tyr Tyr Met Trp

15

<210> 97

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 97

Val Ile Gly Pro Ser Gly Gly Pro Thr Gln Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 98

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 98

Trp Gly Gly Arg Tyr Ser Val Phe Glu Thr

1 5 10

<210> 99

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 99

Arg Ala Ser Gln Ser Val Arg Lys Asn Val Ala

1 5 10

<210> 100

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 100

Gly Ala Ser Thr Arg Ala Thr

15

<210> 101

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 101

Gln Gln Tyr Ser Ser Trp Pro Ala Phe

15

<210> 102

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 102

Arg Tyr Phe Met Gly

15

<210> 103

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 103

Ser Ile Tyr Ser Ser Gly Gly Tyr Thr Ser Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 104

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 104

Arg Trp Arg Asp Gly Thr Phe Asp Tyr

15

<210> 105

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 105

Arg Ala Ser Gln Ser Val Arg Glu Tyr Leu Asn

1 5 10

<210> 106

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 106

Ala Ala Ser Ser Leu Gln Ser

15

<210> 107

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 107

Gln Gln Ser Tyr Ser Thr Arg Phe Thr

15

<210> 108

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 108

Ser Tyr Pro Ile Ser

15

<210> 109

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 109

Lys Ile Ile Pro Ile Phe Gly Thr Thr Asn Tyr Ala Gln Lys Phe Gln

1 5 10 15

Gly

<210> 110

<211> 15

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 110

Pro Asn Arg Pro Tyr Asn Ile Gly Trp His Tyr Tyr Phe Asp Tyr

1 5 10 15

<210> 111

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 111

Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser

1 5 10

<210> 112

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 112

Glu Gly Ser Lys Arg Pro Ser

15

<210> 113

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 113

Ser Ser Tyr Thr Thr Arg Ser Thr Arg Val

1 5 10

<210> 114

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 114

Ser Tyr Ala Met Ser

15

<210> 115

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 115

Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 116

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 116

Asp Gln Asp Glu Gly Arg Ala Asn Asn Trp Trp Ile Pro Pro Gly Gly

1 5 10 15

Arg

<210> 117

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 117

Gln Gly Asp Ser Leu Arg Arg Ser Phe Ala Ser

1 5 10

<210> 118

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 118

Gly Gln Asn Lys Arg Pro Ala

15

<210> 119

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 119

Asn Ser Arg Asp Ala Arg Leu Asn Pro Tyr Ile Leu

1 5 10

<210> 120

<211> 354

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 120

gaagttcaat tgttagagtc tggtggcggt cttgttcagc ctggtggttc tttacgtctt 60

tcttgcgctg cttccggatt cactttctct ccttacatga tgcagtgggt tcgccaagct 120

cctggtaaag gtttggagtg ggtttcttct atctggcctt ctggtggcaa gacttattat 180

gctgactccg ttaaaggtcg cttcactatc tctagagaca actctaagaa tactctctac 240

ttgcagatga acagcttaag ggctgaggac acggccgtgt attactgtgc gagagtgcgg 300

agggggggag ctactgacta ctggggccag ggaaccctgg tcaccgtctc aagc 354

<210> 121

<211> 324

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 121

gacatccaga tgacccagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60

ctctcctgca gggccagtca gagtgttagc agcaacttag gctggtacca gcagaaacct 120

ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccaacc 180

aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240

gaagattttg caacttatta ctgtcagcag tatcagaact ggcccttgct cactttcggc 300

ggagggacca aggtggaaat caaa 324

<210> 122

<211> 357

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 122

gaagttcaat tgttagagtc tggtggcggt cttgttcagc ctggtggttc tttacgtctt 60

tcttgcgctg cttccggatt cactttctct aattactata tgtggtgggt tcgccaagct 120

cctggtaaag gtttggagtg ggtttctgtt atcggtcctt ctggtggccc tactcagtat 180

gctgactccg ttaaaggtcg cttcactatc tctagagaca actctaagaa tactctctac 240

ttgcagatga acagcttaag ggctgaggac acggccgtgt attactgtgc gagatggggt 300

gggaggtact ctgtatttga aacctggggc caagggacaa tggtcaccgt ctcaagc 357

<210> 123

<211> 318

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 123

gacatccaga tgacccagtc tccagccact ctgtctgtgt ctccagggg aagagccacc 60

ctctcctgca gggccagtca gagtgttaga aaaaacgtag cctggtatca gcagaaacct 120

ggccagcctc ccaggctcct catctatggt gcatccacca gggccactgg tgtcccagcc 180

aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag gatgcagcct 240

gaagattttg tagtttatca ctgtcagcag tatagtagct ggccggcgtt cggccagggg 300

accatggtgg aaatcaac 318

<210> 124

<211> 354

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 124

gaagttcaat tgttagagtc tggtggcggt cttgttcagc ctggtggttc tttacgtctt 60

tcttgcgctg cttccggatt cactttctct cgttacttta tgggttgggt tcgccaagct 120

cctggtaaag gtttggagtg ggtttcttct atctattctt ctggtggcta tacttcttat 180

gctgactccg ttaaaggtcg cttcactatc tctagagaca actctaagaa tactctctac 240

ttgcagatga acagcttaag ggctgaggac acggccgtgt attactgtgc gagacggtgg 300

cgagatggca cctttgacta ctggggccag ggaaccctgg tcaccgtctc aagc 354

<210> 125

<211> 321

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 125

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctattggaga cagagtcacc 60

atctcttgcc gggcaagtca gagcgttaga gagtatctaa attggtatca acaaaaacca 120

gggaaagccc ctaaactcct gatctttgct gcatccagtt tgcagagtgg ggtcccatca 180

agattcagtg gcagtggatc tgggacagat ttcactctca ccatcagcag tctgcaacct 240

gaagattttg caacttatta ctgtcaacag agttacagta cccgattcac tttcggccct 300

gggaccaaag tggacatcaa a 321

<210> 126

<211> 372

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 126

caggtacagc tgcagcagtc aggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60

tcctgcaagg cttctggagg caccttcagc agctatccta tcagctgggt gcgacaggcc 120

cctggacaag ggcttgagtg gatgggaaag atcatcccta tctttggtac aacaaactac 180

gcgcagaagt tccagggcag agtcacgatt accgcggacg aatccacgag cactgcctac 240

atggaactga gcagcctgag atctgaggac acggccatat attactgtgc gagccccaat 300

cgaccctata acattggctg gcactactac tttgactact ggggcaaagg aaccctggtc 360

accgtctcct ca 372

<210> 127

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 127

cagtctgtgc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc gatcaccatc 60

tcctgcactg gaaccagcag tgacgttggt ggttataact atgtctcctg gtaccaacaa 120

cacccaggca aagcccccaa actcatgatt tatgagggca gtaagcggcc ctcaggggtt 180

tctaatcgct tctctggctc caggtctggc aacacggcct ccctgacaat ctctgggctc 240

caggctgagg acgaggctga ttattactgc agctcatata caaccaggag cactcgagtc 300

ttcggcggag ggaccaagct gaccgtccta 330

<210> 128

<211> 378

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 128

gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60

tcctgtgcag cctctggatt cacctttagc agctatgcca tgagctgggt ccgccaggct 120

ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180

gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240

ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc aagagatcag 300

gacgaaggta gagcgaacaa ctggtggatc ccccccgggg gtcgctgggg ccaggggaca 360

atggtcaccg tctcgagt 378

<210> 129

<211> 327

<212> DNA

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polynucleotide

<400> 129

tcttctgagc tgactcagga ccctactctg tctgtggccc tgggacagac agtcagaatc 60

acatgccaag gagacagcct ccgaagatct tttgcaagtt ggtaccagaa gaagccagga 120

caggcccctg tacttctcat ctatggtcaa aataagcggc ccgcagggat cccagaccga 180

ttctctggct ccaggtcagg aaactcagct tcgttgacca tcacaggggc tcaggcggaa 240

gatgaggctg actattactg taattcccgc gacgccagac ttaaccctta tatactcttc 300

ggcggtggga ccaagctgac cgtccta 327

<210> 130

<211> 452

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 130

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His

20 25 30

Asp Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Gly Thr Ala Gly Asp Thr Tyr Tyr Pro Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Asp Arg Tyr Ser Pro Thr Gly His Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro

115 120 125

Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr

130 135 140

Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr

145 150 155 160

Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro

165 170 175

Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr

180 185 190

Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn

195 200 205

His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser

210 215 220

Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu

225 230 235 240

Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu

245 250 255

Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val Val Val Asp Val Ser

260 265 270

His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu

275 280 285

Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr

290 295 300

Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn

305 310 315 320

Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro

325 330 335

Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln

340 345 350

Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val

355 360 365

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

370 375 380

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

385 390 395 400

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

405 410 415

Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val

420 425 430

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

435 440 445

Ser Pro Gly Lys

450

<210> 131

<211> 212

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 131

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Asp Asp Phe Ala Thr Tyr Tyr Cys Lys Gln Tyr Ala Asp Tyr Trp Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro

100 105 110

Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr

115 120 125

Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys

130 135 140

Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu

145 150 155 160

Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser

165 170 175

Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala

180 185 190

Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe

195 200 205

Asn Arg Gly Glu

210

<210> 132

<211> 122

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 132

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Val Phe Thr Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Leu Ile Trp Phe Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Arg Gly Gly Gly Tyr Tyr Tyr Tyr Gly Met Asp Val Trp

100 105 110

Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 133

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 133

Ser Tyr Gly Met His

15

<210> 134

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 134

Leu Ile Trp Phe Asp Gly Ser Asn Glu Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 135

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 135

Arg Gly Gly Gly Tyr Tyr Tyr Tyr Gly Met Asp Val

1 5 10

<210> 136

<211> 107

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 136

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp

20 25 30

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 137

<211> 11

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 137

Arg Ala Ser Gln Gly Ile Arg Asn Asp Leu Gly

1 5 10

<210> 138

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 138

Ala Ala Ser Ser Leu Gln Ser

15

<210> 139

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 139

Leu Gln His Asn Ser Tyr Pro Tyr Thr

15

<210> 140

<211> 121

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 140

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly His Glu Trp Met

35 40 45

Gly Trp Ile Ser Ser Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Ser Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ile Ala Ala Arg Gly Tyr Tyr Tyr Gly Met Asp Val Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 141

<211> 5

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 141

Ser Tyr Gly Ile Ser

15

<210> 142

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 142

Trp Ile Ser Ser Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln

1 5 10 15

Gly

<210> 143

<211> 10

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 143

Ala Ala Arg Gly Tyr Tyr Tyr Gly Met Asp

1 5 10

<210> 144

<211> 108

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

polypeptide

<400> 144

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Gly Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 145

<211> 12

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 145

Arg Ala Ser Gln Ser Ile Ser Gly Ser Tyr Leu Ala

1 5 10

<210> 146

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 146

Gly Ala Ser Ser Arg Ala Thr

15

<210> 147

<211> 9

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 147

Gln Gln Tyr Ser Ser Trp Pro Ala Phe

15

<210> 148

<211> 17

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic

peptide

<400> 148

Ser Val Ile Asp Thr Ala Gly Asp Thr Tyr Tyr Pro Gly Ser Val Lys

1 5 10 15

Gly

<---

Claims (33)

1. Pharmaceutical composition for use in the treatment of S. aureus infection, containing an isolated antibody or antigen-binding fragment thereof that specifically binds to S. aureus alpha-toxin (AT), and an isolated antibody or antigen-binding fragment thereof that specifically binds to S. aureus ClfA, wherein CDR1 VH, CDR2 VH, CDR3 VH, CDR1 VL, CDR2 VL and CDR3 VL of the isolated antibody or antigen binding fragment thereof that specifically binds AT corresponds to the amino acid sequences of SEQ ID NO: 69, 70, 71, 1, 2 and 68, respectively . 2. The pharmaceutical composition according to claim 1, where the isolated antibody or its antigen-binding fragment, which specifically binds to AT, contains a heavy chain variable domain having the amino acid sequence of SEQ ID NO:57 and light chain variable domain with amino acid sequence SEQ ID NO:58. 3. The pharmaceutical composition according to claim 1, where the isolated antibody or antigen-binding fragment thereof, which specifically binds to Ab, additionally contains a variant of the Fc region. 4. The pharmaceutical composition according to claim 3, where the isolated antibody or antigen-binding fragment thereof, which specifically binds to AT, contains heavy chain with amino acid sequence SEQ ID NO:130 and light chain with the amino acid sequence SEQ ID NO:131. 5. The composition according to claim 1, where the isolated antibody or antigen-binding fragment thereof, which specifically binds to ClfA, contains: a. CDR1 VH containing the amino acid sequence of SEQ ID NO:133; b. CDR2 VH containing the amino acid sequence of SEQ ID NO:134; c. CDR3 VH containing the amino acid sequence of SEQ ID NO:135; d. CDR1 VL containing the amino acid sequence of SEQ ID NO:137; e. CDR2 VL containing the amino acid sequence of SEQ ID NO:138; And f. CDR3 VL containing the amino acid sequence of SEQ ID NO:139. 6. The composition according to claim 5, where the isolated antibody or antigen-binding fragment thereof, which specifically binds to ClfA, contains VH with amino acid sequence SEQ ID NO:132 and VL with amino acid sequence SEQ ID NO:136. 7. A method of treating an S. aureus infection in a patient, comprising administering to the patient a composition according to any one of claims. 1-6. 8. The method according to claim 7, further comprising administering an antibiotic to the patient. 9. The method according to claim 8, where the antibiotic is penicillin, oxacillin, flucloxacillin or vancomycin. 10. A method for reducing bacterial agglutination of S. aureus and/or the formation of thromboembolic lesions in a patient, including administering to said patient a composition according to any one of claims. 1-6. 11. The method according to claim 10, further comprising administering an antibiotic to the patient. 12. The method according to claim 11, where the antibiotic is penicillin, oxacillin, flucloxacillin or vancomycin. 13. A method for preventing or reducing the severity of sepsis associated with S. aureus in a patient, comprising administering to the patient a composition according to any one of claims. 1-6. 14. The method according to claim 13, further comprising administering an antibiotic to the patient. 15. The method according to claim 14, where the antibiotic is penicillin, oxacillin, flucloxacillin or vancomycin. 16. A method for delaying the onset of sepsis associated with S. aureus infection in a patient, comprising administering to the patient a composition according to any one of claims. 1-6. 17. The method according to claim 16, further comprising administering an antibiotic to the patient. 18. The method according to claim 17, where the antibiotic is penicillin, oxacillin, flucloxacillin or vancomycin.