CN116635024A

CN116635024A - Compositions and methods for treating thyroid eye disorders

Info

Publication number: CN116635024A
Application number: CN202180081949.7A
Authority: CN
Inventors: V·贝迪安; P·哈尔文; T·基塞拉克; A·佘; J·维奥林; Y·赵
Original assignee: Viridian Treatment Co
Current assignee: Viridian Treatment Co
Priority date: 2020-10-14
Filing date: 2021-10-14
Publication date: 2023-08-22
Also published as: CN118059229A; CN116583273A

Abstract

The present application provides antibodies and compositions against IGF-1R, and uses thereof.

Description

Compositions and methods for treating thyroid eye disorders

Cross-reference to related applications:

the present application claims priority from U.S. provisional application No. 63/091,839 to 10, 14, 2021, 5, 21, 63/201,978, 2021, 8, 10, 63/260,130, and 2021, 9, 28, each of which is incorporated herein by reference in its entirety.

The background technology is as follows:

thyroid-related eye diseases (TAOs) (also known as Thyroid Eye Diseases (TED), graves eye diseases or orbital diseases (GO), thyroid toxic abrupt eyes, thyroid dysfunction, and several other terms) are orbital diseases associated with thyroid dysfunction. TAOs fall into two categories. Active TAO, which typically lasts 1 to 3 years, is characterized by a persistent autoimmune/inflammatory response in the orbital soft tissue. Active TAO causes expansion and remodeling of ocular soft tissue. Autoimmune/inflammatory responses of active TAO spontaneously subside and the condition turns into inactive TAO. Inactivity TAO is a term used to describe the long-term/permanent sequelae of active TAO. The etiology of TAO is unknown. TAO is often associated with graves hyperthyroidism, but may also occur as part of other autoimmune conditions that affect the thyroid gland and produce pathology in the orbit and periorbital tissues and rarely in the pre-tibial skin (pre-tibial myxoedema) or fingers (thyroidism). TAO is an autoimmune orbital disease in which the orbit and periocular soft tissues are primarily affected and secondarily affect the eye and vision. In TAO, the eyeball is forced forward (swelled) out of its orbital canal, a phenomenon known as protruding eye (proptosis) or protruding eye (exophalmos), due to inflammation and distention of orbital soft tissue (mainly ocular muscles and fat). Although most cases of TAO do not cause vision loss, this condition can cause exposed keratopathy that endangers vision, troublesome double vision (dipeptida/double vision), and press thyroid dysfunction optic neuropathy. TAO may occur before, contemporaneously with, or after the systemic complications of thyroid dysfunction. Ocular clinical manifestations of TAO include upper eyelid recession, eyelid hysteresis, swelling, redness (erythema), conjunctivitis and raised eyes (protruding or protruding eyes), bulbar conjunctival edema, periorbital edema and altered eye movement with significant functional, social and cosmetic consequences. Many signs and symptoms of TAO, including raised eyes and ocular congestion, are caused by the expansion of orbital adipose tissue and periocular muscles. Part of the cause of the increased adipose tissue volume is new adipocyte development (adipogenesis) within the orbital fat. Accumulation of hydrophilic glycosaminoglycans (mainly hyaluronic acid) within the perimuscular connective tissue between the orbital adipose tissue and the extraocular muscle fibers further dilates the adipose compartment and enlarges the extraocular body. Hyaluronic acid is produced by fibroblasts residing in orbital fat and extraocular muscles, and its synthesis in vitro is stimulated by several cytokines and growth factors including IL-1 beta, interferon-gamma, platelet derived growth factors, thyroid Stimulating Hormone (TSH) and insulin-like growth factor I (IGF-I).

Antibodies that activate insulin-like growth factor I receptor (IGF-IR) are also detected and are involved in the activity TAO. Without being bound by any theory, it is believed that the TSHR and IGF-IR form physiological and functional complexes in orbital fibroblasts, and blocking IGF-IR appears to attenuate IGF-1 and TSH-dependent signaling. Blocking IGF-IR with antibody antagonists has been shown to reduce TSHR and IGF-I dependent signaling and thus interrupt the pathological activity of autoantibodies acting as agonists of either receptor.

IGF-IR is a widely expressed heterotetrameric protein involved in the regulation of proliferation and metabolic function in many cell types. Which is a tyrosine kinase receptor comprising two subunits. IGF-IR alpha contains ligand binding domains, while IGF-IR beta is involved in signaling and contains tyrosine phosphorylation sites.

Current therapies for hyperthyroidism caused by graves' disease are not perfect because of the lack of therapies that target specific underlying pathogenic autoimmune mechanisms of the disease. Even more complex is the treatment of moderate to severe active TAO. Although a deeper understanding of its pathogenesis has been demonstrated in recent years, TAO remains a therapeutic challenge and challenge. There is no approved drug for the treatment of active TAO. Intravenous glucocorticoids (ivGC) and oral glucocorticoids are used to treat patients with moderate to severe active TAO, but the results are less satisfactory. Partial reactions are frequent, and recurrence (rebound) is common after withdrawal. Adverse events occur and many patients eventually require rehabilitation surgery when their pathology has been converted to inactive TAO. Thus, there remains a need to provide alternative therapies for TAO and its related symptoms.

Disclosure of Invention

Embodiments are generally related to IGF-1R antibodies and antigen binding fragments thereof. Certain IGF-1R antibodies and antigen binding fragments inhibit IGF-1R function or block IGF-I mediated biological functions of IGF-1R signaling. In addition, the present invention relates generally to methods for treating thyroid-related eye diseases (TAOs), also known as Thyroid Eye Diseases (TED), graves' eye diseases or orbital diseases (GO), thyrotoxic process eyes, thyroid dysfunction, ocular disorders and other thyroid eye diseases associated with IGF-1R signaling.

In some embodiments, antibodies or antigen binding fragments thereof are provided comprising sequences as provided herein. In some embodiments, the antibody comprises a VL sequence as set forth in SEQ ID NOs 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86; and a VH sequence as set forth in SEQ ID NOs 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83. In some embodiments, the antibody comprises an LCDR sequence as set forth in SEQ ID NOs 17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61, or 81 and an HCDR sequence as set forth in SEQ ID NOs 20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63, or 64; or any combination or variant thereof.

In some embodiments, the antibody or antigen binding fragment thereof comprises V as set forth in SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86 _L A peptide, or any variant thereof. In some embodiments, the antibody or antigen binding fragment thereof comprises V as set forth in SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83 _H A peptide, or any variant thereof.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO:20, 26, 32, 38, 44, 50 or 56; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 21, 27, 33, 39, 45, 51 or 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 22, 28, 34, 40, 46, 52 or 58; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 17, 23, 29, 35, 41, 47 or 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 18, 24, 30, 36, 42, 48 or 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 19, 25, 31, 37, 43, 49, 55 or 81; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 20; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 22; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 17; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 19; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 26; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 28; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 23; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 24; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 25; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 32; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 34; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 29; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 31; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 37; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 44; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 46; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 41; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 43; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 50; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 52; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 47; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 49; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 56; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 58; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 55; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 62; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 64; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 59; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 61; or a variant of any of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 81; or a variant of any of the foregoing.

In some embodiments, the antibody comprises V as set forth in SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86 _L A sequence, or a variant thereof. In some embodiments, the antibody comprises V as set forth in SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83 _H A sequence, or a variant thereof.

In some embodiments, the antibody comprises the sequence of SEQ ID NOS 65 to 72, 78, 82 or 85, or variants thereof.

In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 3 and a heavy chain comprising the amino acid sequence of SEQ ID NO. 83. In some embodiments, the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO. 13 and a heavy chain variable region having the amino acid sequence of SEQ ID NO. 14.

In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain having the amino acid sequence of SEQ ID NO. 92.

In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain having the amino acid sequence of SEQ ID NO. 94.

In some embodiments, the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain having the amino acid sequence of SEQ ID NO. 95.

In some embodiments, variants of any of the antibodies provided herein are provided, as long as the CDRs remain constant compared to the parent (non-variant) sequences provided herein.

In some embodiments, the antibody comprises an Fc region. In some embodiments, the Fc region is as set forth in SEQ ID NOS 75 to 77, 84, 87, 88, 89 or 90. In some embodiments, the Fc region is as set forth in SEQ ID NO. 75. In some embodiments, the Fc region is as set forth in SEQ ID NO. 76. In some embodiments, the Fc region is as set forth in SEQ ID NO: 77. In some embodiments, the Fc region is as set forth in SEQ ID NO. 84. In some embodiments, the Fc region is as set forth in SEQ ID NO. 87. In some embodiments, the Fc region is as set forth in SEQ ID NO. 88. In some embodiments, the Fc region is as set forth in SEQ ID NO. 89. In some embodiments, the Fc region is as set forth in SEQ ID NO. 90.

In some embodiments, pharmaceutical compositions comprising an antibody as provided herein are provided.

In some embodiments, there is provided a method of treating or lessening the severity of thyroid-associated eye disease (TAO) or a symptom thereof, comprising administering to a subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of reducing the Clinical Activity Score (CAS) of thyroid-related eye disease (TAO) in a subject, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of reducing a) the relief of the eye by at least 2mm and b) the reduction of Clinical Activity Score (CAS) in a subject suffering from thyroid-related eye disease (TAO), the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject, the method comprising administering to a subject an antibody as provided herein or a pharmaceutical composition comprising the same, wherein treatment with said antibody (i) reduces the bulge eye of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

In some embodiments, methods of improving the quality of life of a subject suffering from thyroid-related eye disease (TAO, also known as graves 'eye disease/graves' orbit disease) are provided, comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of treating or lessening the severity of a glandular eye disease (TAO) in a subject, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, methods of increasing IGF-1R internalization on a cell are provided, the methods comprising contacting the cell with an antibody or pharmaceutical composition comprising the same as provided herein.

In some embodiments, there is provided a method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, there is provided a method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml, or 105 μg/ml at least 1, 2, or 3 weeks after administration.

In some embodiments, there is provided a method of inhibiting IGF-1 induced receptor autophosphorylation in a cell by at least 95%, 96%, 97%, 98%, or 99% or 100%, the method comprising contacting the cell with an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, embodiments are provided for any of the methods provided herein, wherein the antibody or antigen-binding fragment thereof is administered in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable diluent or excipient or carrier. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for treating TAO. In some embodiments, the pharmaceutical composition further comprises a corticosteroid; rituximab (rituximab) or other anti-CD 20 antibody; tobalizumab or other anti-IL-6 antibodies; or selenium, infliximab or other anti-tnfα antibodies or Thyroid Stimulating Hormone Receptor (TSHR) inhibitors.

Drawings

Fig. 1 shows NHP (non-human primate) serum concentrations of various antibodies and embodiments as provided herein.

Figure 2 shows various properties of antibodies as provided herein.

Fig. 3 illustrates various properties of antibodies as provided herein.

Fig. 4 illustrates various properties of antibodies as provided herein.

Fig. 5 illustrates various properties of antibodies as provided herein.

Fig. 6 illustrates various properties of antibodies as provided herein.

Fig. 7 illustrates various properties of antibodies as provided herein.

Fig. 8 illustrates various properties of antibodies as provided herein.

Detailed Description

Provided herein are antibodies that bind to and modulate the activity of IGF-1R. Antibodies can be used, for example, to treat thyroid eye disease.

As used herein, "thyroid-related eye disease" (TAO), "thyroid eye disease" (TED), "graves eye disease" or "graves orbital disease" (GO) refer to the same disorder or condition and are used interchangeably. It is all referring to inflammatory orbital pathology associated with some autoimmune thyroid disorders, most commonly with "graves 'disease" (GD), but sometimes with other diseases, such as Hashimoto's thyroiditis.

The terms "bulge" and "protrusion (exophthalmos)", also known as the eye protrusion (exophthalmos/exophalmia/exomobits), refer to the anterior protrusion, displacement, bulge or protuberance of an organ. As used herein, the term refers to a forward protrusion, displacement, bulge or protuberance of the eye forward from the orbit. Some of those skilled in the art recognize that the eyes and the eyes have the same meaning and are generally used interchangeably, while others recognize subtle differences in their meaning. Some people use the raised eyes to refer to severe raised eyes; or endocrine related raised eyes. Other people use the term protruding eye when describing a protruding eye in relation to the eye of a subject, e.g. with TAO (TED or GO).

As used herein, the terms "convex eye" and "convex eye" are used interchangeably and refer to an anterior protrusion, displacement, bulge, or protuberance of an eye anteriorly away from the orbit. Any increase in soft tissue content of the orbit, occurring laterally or posteriorly, will displace the eyeball anteriorly due to the rigid bone structure of the orbit with only the anterior opening for dilation. The protruding or protruding eyes may be the result of several disease processes including infection, inflammation, tumors, wounds, cancer metastasis, endocrine lesions, vascular diseases, and external orbital lesions. TAO (TED or GO) is currently recognized as the most common ocular cause in adults. The protruding eyes may be bilateral, as is common in TAO (TED or GO); or unilateral (as is common in orbital tumors).

The extent of the presbyopia can be measured using, for example, a presbyopia meter (an instrument for measuring the extent of forward displacement of the eye). The device allows for measurement of the anterior distance of the lateral orbital margin to the anterior cornea. Computed Tomography (CT) scanning and Magnetic Resonance Imaging (MRI) can also be used to evaluate the extent of a raised eye or eyes. CT scanning is an excellent imaging modality for diagnosing TAO. In addition to allowing for the visualization of the enlarged extraocular muscles, CT scanning also provides the surgeon or clinician with a mapping of the bone anatomy of the orbit when orbit decompression is desired. MRI provides excellent imaging of the orbital contents through its multiplanar and inherent contrast capabilities without the radiation exposure associated with CT scan studies. MRI provides a better imaging of the optic nerve, orbital fat, and extraocular muscles, but CT scanning provides a better view of the bone architecture of the orbit. Orbital ultrasonography may also be used to diagnose and evaluate TAOs because it can be performed quickly and with a high degree of confidence. The high reflectivity and enlargement of the extraocular muscles are readily assessed, and continuous ultrasound examination can also be used to assess the progression or stability of ocular disease. Based on the technology currently available or that will become available in the future, one of ordinary skill in the art will be able to determine the best mode for diagnosing and assessing the extent of a bulge or eye-bump.

As used herein, the term "antibody" refers to any form of antibody that exhibits the desired biological activity. Thus, it is used in its broadest sense and specifically covers, but is not limited to, monoclonal antibodies (including full length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), humanized antibodies, fully human antibodies, chimeric antibodies, and camelized single domain antibodies. A "parent antibody" is an antibody obtained by exposing the immune system to an antigen prior to modification of the antibody for its intended use (e.g., humanization of the antibody for use as a human therapeutic antibody).

As used herein, unless otherwise indicated, "antibody fragment" or "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to an antigen bound by a full-length antibody, e.g., a fragment that retains one or more CDR regions. Examples of antibody binding fragments include, but are not limited to, fab ', F (ab') ₂ And Fv fragments; a bispecific antibody; a linear antibody; single chain antibody molecules, such as sc-Fv; nanobodies and multispecific antibodies formed from antibody fragments.

"Fab fragment" comprises the C of a light chain and a heavy chain _H 1 and a variable region. The heavy chain of a Fab molecule is unable to form disulfide bonds with another heavy chain molecule.

The "Fc" region contains two C comprising antibodies _H 1 and C _H 2 domain. Two heavy chain fragments are linked to C by two or more disulfide bonds _H The hydrophobic interactions of the 3 domains remain together.

In some embodiments, an antibody or antigen fragment herein comprises an Fc region. In some embodiments, the Fc region comprises a mutation that increases the half-life of the antibody when bound to the Fc region. In some embodiments, the Fc region comprises the S228P, L235E, M Y, S254T, T256E, M428L, N434S, L234F, P331S mutation or any combination thereof. In some embodiments, the Fc region comprises M252Y, S T and T256E mutations. Non-limiting examples of Fc regions comprising M252Y, S T and T256E mutations (collectively, "YTE mutations") can be found in the sequence of SEQ ID NO:89. In some embodiments, the Fc region comprising a YTE mutation comprises the sequence of SEQ ID NO:90, which differs from SEQ ID NO:89 by the presence of a C-terminal lysine (K) residue. Numbering of the Fc region may be performed according to the Kabat numbering system for the Fc region.

In some embodiments, the Fc region comprises S228P and L235E mutations. In some embodiments, the antibodies comprise L234F, L E and P331S mutations. In some embodiments, the Fc region comprises M252Y, S254T, T256E, S P and L235E mutations. In some embodiments, the Fc region comprises the S228P, L235E, M428L and N434S mutations. In some embodiments, the Fc region comprises M428L and N434S mutations. In some embodiments, the Fc region comprises L234F, L235E, P331S, M Y, S254T and T256E mutations. Mutations in the Fc region are also described in US2007041972A1, EP2235059B1, U.S. Pat. No. 8,394,925 and Mueller et al, mol Immunol 1997, month 4; 34 441-52, each of which is incorporated herein by reference in its entirety. The numbering referred to herein refers to the Kabat numbering system for the Fc region.

In some embodiments, the Fc region comprises a sequence selected from the group consisting of:

"Fab' fragment" contains one light chain and one heavy chain and contains V _H Domain and C _H 1 domain and C _H 1 and C _H 2 such that an interchain disulfide bond can be formed between the two heavy chains of two Fab 'fragments to form F (ab') ₂ A molecule.

“F(ab') ₂ Fragments "comprise two light chains and two C-containing fragments _H 1 and C _H ² Heavy chains of a portion of the constant region between domains, such that an interchain disulfide bond is formed between the two heavy chains. Thus, F (ab') ₂ Fragments consist of two Fab' fragments held together by disulfide bonds between the two heavy chains.

The "Fv region" comprises variable regions from the heavy and light chains, but lacks constant regions.

The term "single chain Fv" or "scFv" antibody refers to a V comprising an antibody _H And V _L Antibody fragments of domains, wherein these domains are present in a single polypeptide chain. Generally, fv polypeptides are further described in V _H And V is equal to _L The domains contain polypeptide linkers between them that enable scF v to form the desired structure for antigen binding. For reviews of scFv, see Pluck thun (1994) PHARMACOLOGY OF MONOCLONAL ANTIBODIES, volume 113, rosenburg and Moore, springer-Verlag, new York, pages 269-315. See also International patent application publication No. WO 88/01649 and U.S. Pat. Nos. 4,946,778 and 5,260,203.

"domain antibodies" are immunologically functional immunoglobulin fragments that contain only the variable region of a heavy chain or the variable region of a light chain. In some cases, two or more V _H The region is covalently linked to a peptide linker to produce a bivalent domain antibody. Two V of bivalent domain antibody _H The regions may target the same or different antigens.

A "bivalent antibody" comprises two antigen binding sites. In some cases, the two binding sites have the same antigen specificity. However, the bivalent antibody may be bispecific (see below).

In certain embodiments, monoclonal antibodies herein also include camelized single domain antibodies. See, e.g., muyldermans et al (2001) Trends biochem. Sci.26:230; reichmann et al (1999) J.Immunol.methods 231:25; WO 94/04678; WO 94/25591; U.S. patent No. 6,005,079. In one embodiment, the present invention provides a composition comprising two V _H Single domain antibodies to domains, the two V' s _H The domains have modifications such that a single domain antibody is formed.

As used herein, the term "bispecific antibody" refers to a small antibody fragment having two antigen binding sites, said fragment being in the same polypeptide chain (V _H -V _L Or V _L -V _H ) Comprising a light chain variable domain linked (V _L ) Heavy chain variable domain (V) _H ). By using a linker that is so short that it is not possible to pair between two domains on the same strand, the domains are forced to pair with the complementary domain of the other strand, creating two antigen binding sites. Bispecific antibodies are more fully described in, for example, EP 404,097; WO 93/11161; and Holliger et al, (1993) Proc.Natl.Acad.Sci.USA 90:6444-6448. For reviews of engineered antibody variants, see generally Holliger and Hudson (2005) Nat. Biotechnol.23:1126-1136.

Typically, a variant antibody or antigen-binding fragment of an antibody provided herein retains at least 10% of its IGF-1R binding activity when expressed as activity on a molar basis (when compared to the modified parent antibody). In some embodiments, a variant antibody (or antigen fragment thereof) or antigen binding fragment of an antibody provided herein retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of IGF-1R binding affinity compared to the parent antibody. As described herein, it is also contemplated that an antibody or antigen-binding fragment of the invention may include conservative or non-conservative amino acid substitutions that do not substantially alter its biological activity, which may also be referred to as a "conservative variant" or "functional conservative variant" of the antibody.

By "isolated antibody" is meant a purified state of the bound compound, in this context meaning that the molecule is substantially free of other biomolecules, such as nucleic acids, proteins, lipids, carbohydrates, or other materials, such as cell debris and growth media. In general, the term "isolated" is not intended to refer to the complete absence of the material or the absence of water, buffer or salt unless its content substantially interferes with the experimental or therapeutic use of the binding compounds as described herein.

The term "monoclonal antibody" as used herein refers to a substantially homogeneous population of antibodies, i.e., the amino acid sequences of the antibody molecules comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. In contrast, conventional (polyclonal) antibody preparations typically include numerous different antibodies having different amino acid sequences in their variable domains, particularly their CDRs, which are typically specific for different epitopes. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the invention may be made by the hybridoma method first described by Kohler et al (1975) Nature 256:495 or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). "monoclonal antibodies" can also be isolated from phage antibody libraries using techniques described, for example, in Clackson et al (1991) Nature 352:624-628 and Marks et al (1991) J.mol.biol.222:581-597. See also Presta (2005) J.allergy Clin.Immunol.116:731.

As used herein, a "chimeric antibody" is an antibody having a variable domain from a first antibody and a constant domain from a second antibody, wherein the first and second antibodies are from different species. (U.S. Pat. No. 4,816,567 and Morrison et al, (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855). Typically, the variable domain is obtained from an antibody from a laboratory animal of a rodent ("parent antibody") and the constant domain sequence is obtained from a human antibody, such that the resulting chimeric antibody will be less likely to elicit an adverse immune response in a human subject than the parent (e.g., rodent) antibody.

As used herein, the term "humanized antibody" refers to a form of antibody that contains sequences from human and non-human (e.g., murine, rat) antibodies. In general, a humanized antibody will comprise substantially all of at least one and typically two variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the Framework (FR) regions are those of a human immunoglobulin sequence. The humanized antibody may optionally comprise at least a portion of a human immunoglobulin constant region (Fc).

The term "fully human antibody" refers to an antibody comprising only human immunoglobulin sequences. Fully human antibodies may contain murine carbohydrate chains if produced in mice, in mouse cells, or in hybridomas derived from mouse cells. Similarly, "mouse antibody" refers to an antibody that comprises only mouse immunoglobulin sequences. Alternatively, a fully human antibody may contain a rat carbohydrate chain if produced in a rat, a rat cell, or a hybridoma derived from a rat cell. Similarly, "rat antibody" refers to an antibody comprising only rat immunoglobulin sequences.

Typically, the basic antibody structural units comprise tetramers. Each tetramer includes two identical pairs of polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 amino acids or more that is primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector function. Generally, human light chains are classified as kappa light chains and lambda light chains. Furthermore, human heavy chains are generally classified as μ, δ, γ, α or ε, and isotypes of antibodies are defined as IgM, igD, igG, igA and IgE, respectively. Within the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, wherein the heavy chain further includes a "D" region having about more than 10 amino acids. See generally Fundamental ImmunologyChapter 7 (Paul, W., code, 2 nd edition, raven Press, N.Y. (1989)).

The variable regions of each light chain/heavy chain pair form an antibody binding site. Thus, in general, an intact antibody has two binding sites. Except in bifunctional or bispecific antibodies, the two binding sites are generally identical.

Typically, the variable domains of the heavy and light chains comprise three hypervariable regions, also known as Complementarity Determining Regions (CDRs), which are located within relatively conserved Framework Regions (FR). The CDRs are typically aligned by the framework regions so as to be able to bind to a specific epitope. In general, from N-terminus to C-terminus, the light and heavy chain variable domains comprise FR1, CDR1, FR2,CDR2, FR3, CDR3 and FR4. The assignment of amino acids to domains is generally based onSequences of Proteins of Immunological InterestKabat et al; national Institutes of Health, bethesda, md.; 5 th edition; NIH publication No. 91-3242 (1991); kabat (1978) adv. Prot. Chem.32:1-75; kabat et al, (1977) J.biol. Chem.252:6609-6616; chothia et al, (1987) J mol. Biol.196:901-917 or Chothia et al, (1989) Nature 342:878-883.

As used herein, the term "hypervariable region" refers to the amino acid residues in an antibody that are responsible for antigen binding. The hypervariable region comprises amino acid residues from the "complementarity determining region" or "CDR" (i.e., residues 24-34 (CDRL 1), 50-56 (CDRL 2) and 89-97 (CDRL 3) in the light chain variable domain and residues 31-35 (CDRH 1), 50-65 (CDRH 2) and 95-102 (CDRH 3) in the heavy chain variable domain; kabat et al, (1991) Sequences of Proteins ofImmunological Interest, 5 th edition Public Health Service, national Institutes of Health, bethesda, md.) and/or those residues from the "hypervariable loop" (i.e., residues 26-32 (CDRL 1), 50-52 (CDRL 2) and 91-96 (CDRL 3) in the light chain variable domain and 26-32 (CDRH 1), 53-55 (CDRH 2) and 96-101 (CDRH 3) in the heavy chain variable domain; chothia and Lesk (1987) J.mol.917. Biol.196). As used herein, the term "framework" or "FR" residues refer to those variable domain residues other than the hypervariable region residues defined herein as CDR residues. CDRs provide most of the contact residues for the antibody to bind to an antigen or epitope. CDRs of interest may be derived from donor antibody variable heavy and light chain sequences and include analogs of naturally occurring CDRs that also share or retain the same antigen binding specificity and/or neutralizing capacity as the donor antibody from which they are derived.

In some embodiments, the antibody may be in the form of: full length antibodies, single domain antibodies, recombinant heavy chain only antibodies (VHH), single chain antibodies (scFv), shark heavy chain only antibodies (VNAR), micro proteins (cysteine knot proteins, knottins), ankyrin repeat proteins (DARPin); tetranectin (Tetranectin); affibodies (affibodies); transbody (trans body); anti-carrier protein (Anticalin); adanetin (AdNectin); kefir (Affilin); microsomes (Microbody); a peptide aptamer; altrex (alterase); a plastic antibody; fei Luoti (phylomer); stradobody (stradobody); giant body (maxibody); ivermectin (eviody); fenoome (fynomer), armadillo-repeat, kunitz domain, avermectin (avimer), apremimer (atrimer), pran Luo Ti (probody), immunomer, qu Aoshan anti (triomab), troyite (troybody); pepbody (pepbody); vaccinal bodies (vaccinal bodies), monoclonal antibodies (UniBody); affine mo (Affimer), duoBody, fv, fab, fab ', F (ab') 2, peptidomimetic molecules, or synthetic molecules, as described in U.S. patent or patent publication nos. US 7,417,130, US 2004/132094, US 5,831,012, US 2004/0234334, US 7,250,297, US 6,818,418, US 2004/209443, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004/146938, US 2004/157209, US 6,994,982, US 6,794,144, US 2010/239633, US 7,803,907, US 2010/119446, and/or US 7,166,697, each of which is incorporated herein by reference in its entirety. See also Storz mabs.2011 for 5 to 6 months; 3 (3): 310-317, which are incorporated herein by reference.

The term "antigen" as used herein means any molecule capable of producing or binding to an antibody, either directly or indirectly. Included within the definition of "antigen" are nucleic acids encoding a protein. An "antigen" may also refer to a binding partner for an antibody. In some embodiments, the antigen is IGF-1R protein expressed on the surface of cells. In some embodiments, the cell is an intact cell. Intact cells are cells that have not been lysed or ruptured in the presence of a detergent or other agent. Cells that have been treated with detergents or other agents that disrupt or punch in the cell membrane are not intact cells. For example, provided herein are methods for producing antibodies that bind to IGF-1R proteins, comprising culturing cells comprising a nucleic acid molecule encoding an IGF-1R antibody.

As used herein, "specific binding" or "immunospecific binding" or "immunospecificallyBinding "refers to the binding of an antibody to a predetermined antigen (e.g., IGF-1R) or an epitope presented on the antigen. In some embodiments, the antibody is at 10 ^-7 Dissociation constant (K) of M or less _D ) K bound to a predetermined antigen _D K which binds to a non-specific antigen other than the predetermined antigen (e.g. BSA, casein or another non-specific polypeptide) _D At least twice as small. The phrases "antibody that recognizes IGF-1R" and "antibody specific for IGF-1R" are used interchangeably herein with the term "antibody that immunospecifically binds to IGF-1R". The present invention is directed to IGF-1R. The degree of specificity necessary for an anti-IGF-1R antibody may depend on the intended use of the antibody and, at any rate, is defined by its suitability for the intended purpose. In some embodiments, an antibody or binding compound derived from an antigen binding site of an antibody in a contemplated method binds to its antigen (IGF-1R) with at least twice, at least ten times, at least 20 times, or at least 100 times the affinity to any other antigen.

Methods for determining mAb specificity and affinity by competitive inhibition can be found in Harlow, et al, antibodies: A Laboratory Manual, cold Spring Harbor Laboratory Press, cold Spring Harbor, n.y., 1988), coligan et al, eds, current Protocols in Immunology, greene Publishing assoc.and Wiley Interscience, n.y. (1992,1993) and Muller, meth. Enzymol.92:589 601 (1983), which are incorporated herein by reference in their entirety.

The term "homolog" means a protein sequence that has between 40% and 100% sequence homology or identity to a reference sequence. The percent identity between the two peptide chains can be determined by pairwise alignment using the default settings of the AlignX module of Vector ntiv.9.0.0 (Invitrogen corp., carslbad, calif.). In some embodiments, the antibody or antigen binding fragment thereof has at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homology or identity to a sequence described herein. In some embodiments, the antibody has conservative substitutions compared to the sequences described herein. Exemplary conservative substitutions are depicted in table 1 and are encompassed within the scope of the disclosed subject matter. Conservative substitutions may be present in the framework regions or in the antigen binding sites, provided that they do not adversely affect the properties of the antibody. Substitutions may be made to improve antibody properties, such as stability or affinity. Conservative substitutions will result in molecules having functional and chemical characteristics similar to those of such modifications. Exemplary amino acid substitutions are shown in the table below.

In some embodiments, variants of the proteins and peptides provided herein are provided. In some embodiments, the variant comprises a substitution, deletion, or insertion. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) substitutions. As described herein, a substitution may be a conservative substitution. In some embodiments, the substitution is non-conservative. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) deletions. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) insertions. In some embodiments, substitutions, deletions, or insertions are present in CDRs provided herein. In some embodiments, the substitution, deletion, or insertion is not present in a CDR provided herein.

The term "in combination with … …" as used herein means that the described agents can be administered to an animal or subject together in admixture, simultaneously as a single agent, or sequentially in any order as a single agent.

Techniques for generating antibodies to small peptide sequences that recognize and bind to those sequences that are in free or conjugated form or, in the case of larger proteins, appear as native sequences are well known in the art. Such antibodies include murine, murine-human, and human-human antibodies produced by hybridoma or recombinant techniques known in the art. Antibodies may also be produced in humans, mice, sheep, rats, rabbits, sharks, camels or chickens. In some embodiments, the antibody is produced in chicken. Antibodies can also be produced in other small animals.

The term "epitope" means a portion of any molecule capable of being recognized and bound by an antibody at one or more of the antigen binding regions of an Ab. Epitopes are generally composed of chemically active surface groups of molecules such as amino acids or sugar side chains, and have specific three-dimensional structural features as well as specific charge characteristics. Examples of epitopes include, but are not limited to, residues described herein that form IGF-1R epitopes. In some embodiments, the epitope is present only in non-denatured proteins. In some embodiments, the epitope is present only in the denatured protein.

In some embodiments, the source of DNA encoding the non-human antibody includes an antibody-producing cell line, such as a hybrid cell line commonly known as a hybridoma.

Hybrid cells are formed by fusion of non-human antibody-producing cells (typically spleen cells of animals immunized with a native or recombinant antigen, or peptide fragment of an antigen protein sequence). Alternatively, the non-human antibody-producing cells may be B lymphocytes obtained from the blood, spleen, lymph nodes or other tissues of an antigen-immunized animal.

The second fusion partner providing immortal function may be lymphoblastic or plasmacytoma or myeloma cells, which are not themselves antibody-producing cells, but are malignant. Fusion partner cells include, but are not limited to, hybridomas SP2/0-Ag14, abbreviated as SP2/0 (ATCC CRL 1581) and myeloma P3X63Ag8 (ATCC TIB 9) or derivatives thereof. See, e.g., ausubel, infra, harlow, infra, and Colligan, infra, the contents of which are incorporated herein by reference in their entirety.

Antibodies can be produced according to the examples provided herein. Once the sequences are known, antibodies can also be generated according to known methods. Antibodies can also be converted to different types, such as to human IgG, and the like. By converting an antibody to a human antibody, the human subject should not recognize the antibody as an foreign antibody. Conversion of non-human IgG antibodies to human IgG antibodies is well known and can be routinely performed once the native sequence is known. As discussed herein, antibodies can be modified according to known methods. Such methods are described, for example, in Riechmann L, clark M, waldmann H, winter G (1988) Reshaping human antibodies for treatment. Nature 332 (6162): 332-323; twushita N, park M, pakabunto K, ong K, avdalovic A, fu H, jia, V.squez M, kumar S. (2004). Antibody-producing cells that promote nucleotide sequences encoding the antigen-binding region of a chimeric antibody can also be produced by transforming non-human (e.g., primate) or human cells. For example, antibody-producing B lymphocytes can be infected with a virus such as Epstein-Barr virus (Kozbor et al, immunol. Today 4:72:79 (1983)) and transformed to give immortalized antibody-producing cells. Alternatively, B lymphocytes may be transformed by providing a transgene or a transgene product, as is well known in the art. See, e.g., ausubel, infra, harlow, infra, and Colligan, infra, the contents of which are incorporated herein by reference in their entirety. Cell fusion is achieved by standard procedures well known to those skilled in the immunological arts. Fusion partner cell lines and methods for fusion and selection of hybridomas and selection of mabs are well known in the art. See, for example, ausubel, harlow, infra, and Colligan, infra, the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the antibody is a MAb that binds to IGF-1R. In some embodiments, the antibody binds to an amino acid of an epitope of IGF-1R.

In some embodiments, the antibody comprises a sequence as provided herein.

The sequence of the antibody may be modified to produce a human IgG antibody. The transformation of the sequences provided herein may be modified to produce other types of antibodies. The CDRs may also be linked to other antibodies, proteins, or molecules to produce antibody fragments that bind to IGF-1R. Which may be in the form of an antibody drug conjugate ("ADC"), a multispecific molecule, or a chimeric antigen receptor. The CDR and antibody sequences provided herein are also humanized or made fully human according to known methods. The sequences may also be made into chimeric antibodies as described herein.

In some embodiments, the antibody comprises an amino acid sequence comprising a sequence provided herein or a fragment thereof. In some embodiments, the antibody comprises one or more amino acid sequences, antigen binding fragments thereof, or human IgG variants thereof, as provided herein. "human IgG variant thereof" refers to an antibody that has been modified to human IgG when the starting antibody is not a human IgG antibody.

As described herein, the production of antibodies having known sequences is conventional and can be performed by any method. Thus, in some embodiments, nucleic acids encoding antibodies or fragments thereof are provided. In some embodiments, the nucleic acid encodes a sequence provided herein. Antibodies may also be modified to chimeric or human antibodies. Antibodies may also be used in the form of injectable pharmaceutical compositions. Also as described herein, the antibody may be an isolated antibody or an engineered antibody.

In some embodiments, there is provided a "derivative" of an antibody, fragment, region or derivative thereof, including those proteins encoded by genes truncated or modified to produce a molecular species functionally similar to an immunoglobulin fragment. Modifications include, but are not limited to, the addition of genetic sequences encoding cytotoxic proteins such as plant and bacterial toxins. Modifications may also include reporter proteins, such as fluorescent or chemiluminescent tags. Fragments and derivatives may be produced in any manner.

The identification of these antigen binding regions and/or epitopes recognized by the abs described herein provides information required to generate additional monoclonal antibodies with similar binding characteristics and therapeutic or diagnostic utility as compared to embodiments of the application.

The nucleic acid sequence encoding an antibody described herein can be genomic DNA or cDNA, or RNA (e.g., mRNA) encoding at least one of the variable regions described herein. A suitable alternative to using chromosomal gene segments as a source of DNA encoding the V region antigen binding fragments is to use cDNA to construct chimeric immunoglobulin genes, e.g., as reported by Liu et al (Proc.Natl.Acad.Sci., USA 84:3439 (1987) and J.immunology 139:3521 (1987)), which references are incorporated herein by reference in their entirety. The use of cDNA requires the combination of gene expression elements and genes appropriate for the host cell in order to synthesize the desired protein. The use of cDNA sequences is preferred over genomic sequences (which contain introns) because cDNA sequences can be expressed in bacteria or other hosts lacking a suitable RNA splicing system.

For example, a cDNA encoding a V region antigen binding fragment capable of detecting, binding to, or neutralizing IGF-1R antigen may be provided using known methods based on the use of the amino acid sequences provided herein. Because the genetic code is degenerate, more than one codon may be used to encode a particular amino acid (Watson et al, see below). One or more different oligonucleotides can be identified using the genetic code, each of which will be capable of encoding an amino acid. The probability that a particular oligonucleotide will actually constitute an actual XXX coding sequence can be estimated by considering the abnormal base pairing relationships in eukaryotic or prokaryotic cells expressing the antibody or fragment and the frequency with which particular codons (encoding particular amino acids) are actually used. Such "rules of codon usage" are disclosed by Lathes et al, J.molecular.biol.183:1.12 (1985). Using the "codon usage rules" of Lathes, a single oligonucleotide or collection of oligonucleotides containing a theoretical "most likely" nucleotide sequence capable of encoding an antibody variable or constant region sequence was identified.

The variable regions described herein can be combined with any type of constant region, including human constant regions or murine constant regions. The human genes encoding the constant (C) regions, fragments and regions of antibodies can be derived from the human fetal liver repertoire by known methods. The human C region gene may be derived from any human cell, including human cells that express and produce human immunoglobulins. Human C _H The regions may be derived from any of the known classes or isotypes of the human H chain, including γ, μ, α, δ, or ε, and subtypes thereof, such as G1, G2, G3, and G4. Since the H chain isotype is responsible for various effector functions of antibodies, C _H The selection of the region will be guided by the desired effector function, such as complement fixation, or activity in Antibody Dependent Cellular Cytotoxicity (ADCC). Preferably C _H The region is derived from γ1 (IgG 1), γ3 (IgG 3), γ4 (IgG 4) or μ (IgM). Human C _L The region may be derived from human L chain isotypes, kappa or lambda. In some embodiments, the antibody comprises an Fc domain. In some embodiments, the Fc domain comprises a mutation that increases the half-life of the antibody. In some embodiments, the Fc domain comprises a processSuch as those described in U.S. patent No. 7,670,600, which is incorporated by reference herein in its entirety. In some embodiments, the constant region comprises a mutation at amino acid residue 428 relative to a wild-type human IgG constant domain, said positions being numbered according to the EU numbering index of Kabat. Without being bound by any particular theory, antibodies comprising mutations corresponding to residue 428 may have an extended half-life compared to the half-life of IgG having wild-type human IgG constant domains. In some embodiments, the mutation is to replace a natural residue with threonine, leucine, phenylalanine, or serine. In some embodiments, the antibody further comprises one or more amino acid substitutions at one or more of amino acid residues 251-256, 285-290, 308-314, 385-389, and 429-436, relative to the corresponding wild-type human IgG constant domain, which amino acid residues are numbered according to the Kabat EU numbering index. Specific mutations or substitutions at these positions are described in U.S. patent No. 7,670,600, which is incorporated herein by reference in its entirety.

Genes encoding the human immunoglobulin C region can be obtained from human cells by standard cloning techniques (Sambrook et al (Molecular Cloning: A Laboratory Manual, 2 nd edition, cold Spring Harbor Press, cold Spring Harbor, N.Y.) (1989) and Ausubel et al, code Current Protocols in Molecular Biology (1987 1993)). Human C region genes are readily obtained from known clones containing genes representing two classes of L chains, five classes of H chains and sub-classes thereof. Such as F (ab') ₂ And chimeric antibody fragments of Fab can be prepared by designing the appropriate truncated chimeric H chain gene. For example, code F (ab') ₂ Chimeric genes of the H chain portion of the fragment will include a CH encoding the H chain ₁ The DNA sequence of the domain and hinge region is followed by a translation stop codon, thereby resulting in a truncated molecule.

In some embodiments, antibodies, murine, human, humanized or chimeric antibodies, fragments and regions of antibodies described herein are prepared by cloning DNA fragments encoding the H and L chain antigen binding regions of IGF-1R antigen-specific antibodies, and ligating these DNA fragments to DNA fragments encoding C-chain antigen binding regions, respectively _H And C _L DNA fragments of the regions are generated to produce murine, human or chimeric immunoglobulin encoding genes.

Thus, in some embodiments, a fusion chimeric gene is formed comprising at least a first DNA fragment encoding an antigen binding region of non-human origin (e.g., functionally rearranged V region) linked by a junction (J) fragment to a second DNA fragment encoding at least a portion of a human C region.

Thus, the cDNA encoding antibody V and C regions, the method of producing antibodies according to some embodiments described herein, involves several steps, as exemplified below: 1. isolating messenger RNA (mRNA) from cell lines producing antibodies to IGF-1R antigen and from optional additional antibodies supplying heavy and light chain constant regions; cloning and generating cDNA therefrom; 2. preparing a full-length cDNA library from purified mRNA, wherein suitable V and/or C region gene fragments of L and H chain genes can be: (i) identifying with a suitable probe, (ii) sequencing and (iii) compatibilizing the C or V gene fragment with another antibody from the chimeric antibody; 3. constructing a complete H or L chain coding sequence by ligating cloned specific V region gene fragments to cloned C region genes as described above; 4. the L and H chains are expressed and produced in selected hosts (including prokaryotic and eukaryotic cells) to provide murine-murine, human-human or human-murine antibodies.

Two coding DNA sequences are said to be "operably linked" if the ligation results in a continuous translatable sequence without a change or disruption of the triple reading frame. A coding sequence is operably linked to a gene expression element if the linkage produces the proper function of the gene expression element to cause expression of the DNA coding sequence.

As used herein and unless otherwise indicated, the term "about" is intended to mean ± 5% of the value it modifies. Thus, about 100 means 95 to 105.

In some embodiments, the presence of an antigen is detected using an antibody described herein. The antibodies of the invention may be used in any device or method to detect the presence of an antigen.

The term "purified" with reference to an antibody refers to an antibody that is substantially free of other materials associated with molecules in its natural environment. For example, the purified protein is substantially free of cellular material or other proteins of the cell or tissue from which it is derived. The term refers to a protein wherein the purity of the isolated protein is sufficient for analysis or is at least 70% to 80% (w/w) pure, at least 80% -90% (w/w) pure, 90-95% pure; and at least 95%, 96%, 97%, 98%, 99% or 100% (w/w) pure formulation. In some embodiments, the antibody is purified.

As an alternative to preparing monoclonal antibody secreting hybridomas, monoclonal antibodies directed against the polypeptides described herein can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phage display library) with the polypeptides described herein, thereby isolating immunoglobulin library members that bind the polypeptides. Techniques and commercially available kits for generating and screening phage display libraries are well known to those of skill in the art. In addition, examples of methods and reagents particularly useful for generating and screening libraries of antibody or antigen binding protein presentations can be found in the literature. Thus, the epitopes described herein can be used to screen for other antibodies that can be used therapeutically, diagnostically, or as research tools.

Antibody conjugates

The antibodies provided herein can also be conjugated to a chemical moiety. The chemical moiety may be, inter alia, a polymer, a radionuclide or a cytotoxic factor. In some embodiments, this may be referred to as an antibody drug conjugate. In some embodiments, the chemical moiety is a polymer that increases the half-life of the antibody molecule in the subject. Suitable polymers include, but are not limited to, polyethylene glycol (PEG) (e.g., PEG having a molecular weight of 2kDa, 5kDa, 10kDa, 12kDa, 20kDa, 30kDa, or 40 kDa), dextran, and monomethoxy polyethylene glycol (mPEG). Lee et al, (1999) (bioconj. Chem. 10:973-981) disclose single chain antibodies conjugated to PEG. Wen et al, (2001) (bioconj. Chem. 12:545-553) disclose conjugated antibodies with PEG attached to a radiometal chelator (diethylenetriamine pentaacetic acid (DTPA)). Examples of chemical moieties include, but are not limited to, antimitotic agents such as calicheamicin (e.g., ozagrimocin), monomethyl auristatin E, maytansine, and the like. Other examples include, but are not limited to, bioactive anti-microtubule agents, alkylating agents, and DNA minor groove binding agents. Other examples are provided herein and below. The chemical moiety may be attached to the antibody through a linking group (maleimide), a cleavable linker, such as a cathepsin cleavable linker (valine-citrulline), and in some embodiments one or more spacers (e.g., para-aminobenzyl carbamate). Without being bound by any particular theory, once the antibody conjugate binds IGF-1R, it can internalize and the chemical moiety can kill the cell or otherwise inhibit its growth. In some embodiments, the cell is a thyroid cell.

The antibodies and antibody fragments of the invention may also be conjugated with labels such as: ⁹⁹ Tc、 ⁹⁰ Y、 ¹¹¹ In、 ³² P、 ¹⁴ C、 ¹²⁵ I、 ³ H、 ¹³¹ I、 ¹¹ C、 ¹⁵ O、 ¹³ N、 ¹⁸ F、 ³⁵ S、 ⁵¹ Cr、 ⁵⁷ To、 ²²⁶ Ra、 ⁶⁰ Co、 ⁵⁹ Fe、 ⁵⁷ Se、 ¹⁵² Eu、 ⁶⁷ CU、 ²¹⁷ Ci、 ²¹¹ At、 ²¹² Pb、 ⁴⁷ Sc、 ¹⁰⁹ Pd、 ²³⁴ th and ⁴⁰ K、 ¹⁵⁷ Gd、 ⁵⁵ Mn、 ⁵² tr and Tr ⁵⁶ Fe。

Antibodies and antibody fragments may also be conjugated with fluorescent or chemiluminescent labels including fluorophores such as rare earth chelators, luciferin and derivatives thereof, rhodamine (rhodomine) and derivatives thereof, isothiocyanates, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde, fluorescamine (fluorescamine), and, ¹⁵² Eu, dansyl (dansyl), umbelliferone (umbelliferone), fluorescein (luciferin), luminol (lumil), isoluminol (isoluminal label), aromatic acridinium ester (aromatic acridinium ester) labels, imidazole labels, acridinium salt (acridinium salt) labels, oxalate labels, jellyfish-labels (aequorin label), 2, 3-dihydro phthalazinedione, biotin/avidin, spin labels and stable free radicals.

Antibody molecules may also be conjugated to cytotoxic factors such as diphtheria toxin, pseudomonas aeruginosa (Pseudomonas aeruginosa) exotoxin a chain, ricin a chain, abrin a chain, mo Disu (modeccin) a chain, alpha-sarcin (alpha-sarcosine), tung (Aleurites fordii) proteins and compounds (e.g., fatty acids), caryophyllin, pokeweed protein (Phytoiacca americana protein) PAPI, PAPII and PAP-S, balsam pear (momordica charantia) inhibitors, curcin, crotonin, soapbark (saponaria officinalis) inhibitors, mitomycin, restrictocin, phenol, and enomycin.

Any method known in the art for conjugating an antibody molecule of the invention to various moieties may be used, including those described by Hunter et al, (1962) Nature 144:945; david et al, (1974) Biochemistry 13:1014; pain et al, (1981) J.Immunol. Meth.40:219; and Nygren, J. (1982) Histochem. And Cytochem. 30:407. Methods of conjugating antibodies are conventional and well known in the art.

Chimeric antigen receptor

The antibodies provided herein can also be incorporated into chimeric antigen receptors ("CARs") that can be used, for example, in CAR-T cells. In some embodiments, the extracellular domain of the CAR can be an antibody as provided herein. In some embodiments, the antibody is in the form of an scFv. CAR-T cells are a type of treatment in which the T cells of the patient are modified so that they will attack cells expressing IGF-1R. T cells are obtained from the blood of a patient. Next, genes for specific receptors for a certain protein that bind to the patient's cells are added in the laboratory. In some embodiments, the receptor binds to IGF-1R using the binding region of the antibodies provided herein. CAR-T cells comprising IGF-1R antibodies can then be used to treat conditions, such as those provided herein.

In some embodiments, provided herein are antibodies (e.g., anti-IGF-1R antibodies). In some embodiments, the antibody is a recombinant antibody that binds to IGF-1R protein. In some embodiments, the IGF-1R protein is a human IGF-1R protein. In some embodiments, the IGF-1R protein recognized by the antibody is in its native (non-denatured) conformation. In some embodiments, the antibody does not specifically bind to a denatured IGF-1R protein. As used herein, the term "recombinant antibody" refers to an antibody that does not naturally occur. In some embodiments, the term "recombinant antibody" refers to an antibody that is not isolated from a human subject.

In some embodiments, the antibody comprises one or more peptides or variants thereof having the sequence:

the columns indicated as antibody sequences contain the VH and VL chains of the antibodies. Where a VH chain is described with an Fc sequence, the Fc sequence may be modified or substituted for a different Fc region as provided herein. However, in some embodiments, an antibody may comprise VH and VL sequences as provided in the tables provided herein. For example, in some embodiments, an antibody comprises one or more VH, HC, LC, or VL having the following sequences (those having constant domains are complete light or heavy chains) or variants thereof:

In some embodiments, the variable light chain as set forth in SEQ ID NO. 13 does not have a C-terminal arginine residue. This is illustrated, for example, in the following sequence:

thus, in some embodiments, where the variable light chain comprises the sequence of SEQ ID NO. 13, it may be substituted with the sequence of SEQ ID NO. 97.

In some embodiments, the heavy chain variable region as set forth in SEQ ID NO. 14 may comprise a C22S substitution. This is illustrated in the following sequence:

thus, in some embodiments, the antibody comprises the VH sequence of SEQ ID NO:96 and the VL sequence of SEQ ID NO:13 or SEQ ID NO: 97.

In some embodiments, the antibody comprises the VH of SEQ ID NO. 14 and the VL sequence of SEQ ID NO. 97.

In some embodiments, the antibody comprises a VL of SEQ ID NO. 98 and a VH of SEQ ID NO. 99. In some embodiments, the antibody comprises the VL of SEQ ID NO. 98 and the VH of SEQ ID NO. 99, having an Fc region comprising M252Y, S254T and T256E mutations. In some embodiments, the antibody comprises the VL of SEQ ID NO. 98 and the VH of SEQ ID NO. 99, having an Fc region comprising the M428L and N434S mutations.

As provided herein, heavy chains may be linked to the Fc region, including those with mutations that may affect the half-life of the antibody. Non-limiting mutations in the Fc region are provided herein.

In the tables provided herein, LC and HC can be described with VH and VL domains with or without constant regions. The constant regions may be replaced as provided herein. The VH and VL regions can be used to form antibodies as provided herein. The VH and VL sequences may be in any form, including (but not limited to) scFv forms wherein the VH and VL regions are linked by a peptide linker. Examples of peptide linkers that can be used to link the various peptides provided herein include (but are not limited to): (GGGGS) _n (SEQ ID NO:73)；(GGGGA) _n (SEQ ID NO: 74) or any combination thereof, wherein each n is independently 1 to 5. In some embodiments, the variable regions are not linked by a peptide linker. In some embodiments, the antibody comprises SEQ ID NO. 1 and SEQ ID NO. 2, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO. 3 and SEQ ID NO. 4, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO 5 and SEQ ID NO 6, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO. 7 and SEQ ID NO. 8, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO 9 and SEQ ID NO 10, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO. 11 and SEQ ID NO. 12, or CDR regions thereof. In some embodiments, the antibody comprises SEQ ID NO. 13 and SEQ ID NO. 14, or CDR regions thereof. At the position of In some embodiments, the antibody comprises SEQ ID NO. 15 and SEQ ID NO. 16, or CDR regions thereof.

In some embodiments, an antibody or antigen binding fragment thereof is provided, wherein the antibody or antibody fragment comprises a peptide selected from the following table.

In some embodiments, the antibody or antibody binding fragment thereof comprises heavy or light chain CDRs with sequences of SEQ ID NOs 17 to 64 and 81. In some embodiments, the antibody or antibody binding fragment thereof comprises a light chain CDR having the sequence of SEQ ID NO:17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61, or 81. In some embodiments, the antibody or antibody binding fragment thereof comprises a heavy chain CDR having the sequence of SEQ ID NO:20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63, or 64.

In some embodiments, the antibody or antibody binding fragment thereof comprises a light chain having LCDR1, LCDR2, and LCDR3, wherein LCDR1 has the sequence of SEQ ID NO:17, 23, 29, 35, 41, 47, 53, or 59, LCDR2 has the sequence of SEQ ID NO:18, 24, 30, 36, 42, 48, 54, or 60, and LCDR3 has the sequence of SEQ ID NO:19, 25, 31, 37, 43, 49, 55, 61, or 81.

In some embodiments, the antibody or antibody binding fragment thereof comprises a heavy chain having HCDR1, HCDR2 and HCDR3, wherein HCDR1 has the sequence of SEQ ID NO:20, 26, 32, 38, 44, 50, 56 or 62, HCDR2 has the sequence of SEQ ID NO:21, 27, 33, 39, 45, 51, 57 or 63, and HCDR3 has the sequence of SEQ ID NO:22, 28, 34, 40, 46, 52, 58 or 64.

The different CDR motifs may be combined in any combination, including those combinations not depicted in the table above. For example, the following embodiments provide non-limiting examples of such combinations.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 17; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 19; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 20; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 22; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 23; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 24; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 25; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 26; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 28; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 29; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 31; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 32; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 34; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 37; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 41; light chain CDR2 has the amino acid sequence of SEQ ID NO. 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 43; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 44; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 46; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 47; light chain CDR2 has the amino acid sequence of SEQ ID NO. 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 49; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 50; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 52; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 53; light chain CDR2 has the amino acid sequence of SEQ ID NO. 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 55; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 58; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 59; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 61; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 62; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 64; or a variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises: (i) A light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 81; and (ii) a heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing.

In some embodiments, the light chain variable region CDR1 is replaced with any of the other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced with any of the other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced with any of the other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced with any of the other light chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced with any of the other light chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced with any of the other light chain CDR3 sequences.

In some embodiments, antibodies or antigen binding fragments thereof or proteins are provided comprising peptides having the sequences as set forth in any one of SEQ ID NOs 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86, and 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83.

In some embodiments, the antibody or antigen binding fragment thereof comprises a sequence or variant of any of the foregoing.

In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO. 65 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:66 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:67 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO. 68 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:69 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO. 70 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:71 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:72 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:78 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO. 82 or a variant of any of the foregoing. In some embodiments, the antibody or antigen binding fragment thereof comprises the sequence of SEQ ID NO:85 or a variant of any of the foregoing.

In some embodiments, V _L And/or V _H The sequences are provided herein. In some embodiments, V _L The sequences are provided as elements of the Light Chain (LC). In some embodiments, V is provided as an element of the Light Chain (LC) _L The sequence is underlined in the LC sequence. In some embodiments, V is provided as an element of a heavy chain (LC) _H The sequence is underlined in the HC sequence.

In some embodiments, an anti-cancer agentThe body or antigen binding fragment thereof comprises V as set forth in SEQ ID NO. 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86 _L A peptide, or any combination thereof. V (V) _L The peptide may comprise a variant of any of these sequences as provided herein.

In some embodiments, the antibody or antigen binding fragment thereof comprises V as set forth in SEQ ID NO. 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83 _H A peptide, or any combination thereof. V (V) _H The peptide may comprise a variant of any of these sequences as provided herein.

In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L A peptide. Wherein V is _H The peptide comprises a sequence as set forth in SEQ ID NO 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83 and V _L The peptide comprises the sequence set forth in SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86.

In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 2 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 1. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 4 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 3. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 6 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 5. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 8 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 7. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO 10 and V _L The peptide comprising a peptide as set forth in SEQ ID NO 9Sequence. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 12 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 11. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 14 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 13. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 16 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 15. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 80 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 79. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO:83 and V _L The peptide comprises the sequence as set forth in SEQ ID NO. 3. In some embodiments, the antibody or antigen-binding fragment thereof comprises V _H Peptides and V _L Peptides, wherein V _H The peptide comprises the sequence as set forth in SEQ ID NO. 14 and V _L The peptide comprises the sequence set forth in SEQ ID NO. 86.

In some embodiments, the antibody or antigen binding fragment thereof comprises an LC peptide as set forth in SEQ ID NOs 1, 3, 5, 7, 9, or 11, or any combination thereof. LC peptides may comprise variants of any of these sequences as provided herein.

In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide as set forth in SEQ ID NO. 2, 4, 6, 8, 10, 12, or 83, or any combination thereof. The HC peptide may comprise a variant of any of these sequences as provided herein.

In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID No. 2, 4, 6, 8, 10, 12 or 83 and the LC peptide comprises a sequence as set forth in SEQ ID No. 1, 3, 5, 7, 9 or 11. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO. 2 and the LC peptide comprises a sequence as set forth in SEQ ID NO. 1. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO. 4 and the LC peptide comprises a sequence as set forth in SEQ ID NO. 3. In some embodiments, HC peptides comprising the sequence set forth in SEQ ID NO. 4 have an additional C-terminal lysine (K) residue. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO. 6 and the LC peptide comprises a sequence as set forth in SEQ ID NO. 5. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO. 8 and the LC peptide comprises a sequence as set forth in SEQ ID NO. 7. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO:10 and the LC peptide comprises a sequence as set forth in SEQ ID NO: 9. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO:12 and the LC peptide comprises a sequence as set forth in SEQ ID NO: 11. In some embodiments, the antibody or antigen binding fragment thereof comprises an HC peptide and an LC peptide, wherein the HC peptide comprises the sequence as set forth in SEQ ID NO:83 and the LC peptide comprises the sequence as set forth in SEQ ID NO: 3.

In addition to these specific combinations, V _H Peptides and V _L Any of the peptides may be combined with one another.

In addition to these specific combinations, any of the HC peptide and LC peptide may be combined with each other.

In some embodiments, the antibody comprises the sequence or antigen binding fragment of ATCC clone PTA-7444. The antibody sequence produced by ATCC clone PTA-7444, including antigen binding fragments thereof, is incorporated herein by reference in its entirety.

In addition, as provided herein, an antibody may be a multispecific antibody in that the antibody has multiple binding regions targeting different proteins or the same protein at different epitopes. In some embodiments, the antibody is a bispecific antibody.

As provided herein, the different peptides described herein (V _H Or V _L ) May be linked by a peptide linker or not and are alternatively a contiguous sequence. In some embodiments, the peptide linker comprises the sequence: (GGGGS) _n (SEQ ID NO:73)；(GGGGA) _n (SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5. The linked peptide form may be represented by V _H -Z-V _L Or V _L -Z-V _H Wherein Z is a peptide linker. In some embodiments, Z is (GGGGS) _n (SEQ ID NO:73)；(GGGGA) _n (SEQ ID NO: 74) or any combination thereof, wherein each n is independently 1 to 5.

As provided herein, an antibody or antigen binding fragment thereof may be a variant of a sequence.

Other examples of antibodies include, but are not limited to, those provided in the following: each of US20160096894A1, EP1399483B1, EP2194067B1, US20040202651A1, US20110229933A1, US8137933B2, US8951790B2, US20190270820A1, US7572897B2, US20090275126A1, EP1959014B1, US20080014203A1, US20080226635A1, US20120076778A1, US20190153071A1, WO2011161119A1, US10611825B2, US20120237507A1, EP2681240B1, US9982036B2, US20180312573A1, EP2681239B1, US20160151487A1, US20190225696A1, WO2017011773A2, US20200023076A1, US20190153471A1, US20190194713A1, WO2020006486A1, US20080112888A1, US20150168424A1, EP2032989B2, US9045536B2, are incorporated herein by reference in their entirety. Other examples of antibodies include, but are not limited to, those provided in the following: each of US8153121B2, EP1469879B1, WO2016064716A1, US20190270820A1, US20180280527A1, US20190225696A1, US7998681B2, US20040202651A1, US20050136063A1, US20090285824A1, US20150274829A1, EP2322550B1, US20060286103A1, US20070071675A1, US20100047239A1, US20130004416A1, US20080112888A1, US20150168424A1, US20100143340A1, US20110014117A1, US20100260668A1, US20100074900A1, US20150017168A1, US20110044980A1, US20130330323A1, US20120263722A1, US20120201746A1, US10519245B2, US20180243432A1, US20170218091A1, US20200115460A1, US20100104645A1, US20120065380A1, EP2970433B1, US20160289341A1, US20160289343A1, US20190293656A1, and the like, each of which is incorporated herein by reference in its entirety.

In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the sequence:

and the light chain comprises the following sequence:

in some embodiments, the heavy chain of SEQ ID NO. 94 comprises a C-terminal lysine residue added to the C-terminal end of SEQ ID NO. 94.

and the light chain comprises the sequence of SEQ ID NO. 93.

In some embodiments, the heavy chain of SEQ ID NO. 95 comprises a C-terminal lysine residue added to the C-terminal end of SEQ ID NO. 95.

In some embodiments, the antibody comprises a heavy chain and a light chain, wherein the heavy chain comprises the sequence of SEQ ID NO. 83 and the light chain comprises the sequence of SEQ ID NO. 3.

In some embodiments, the antibody comprises the VH sequence of SEQ ID NO:96 and the VL sequence of SEQ ID NO:13 or SEQ ID NO: 97. In some embodiments, the antibody comprises the VH of SEQ ID NO. 14 and the VL sequence of SEQ ID NO. 97.

Pharmaceutical composition

In some embodiments, to prepare a pharmaceutical or sterile composition of an anti-IGF-1R antibody or other protein provided herein, an antibody or antigen-binding fragment thereof or other protein provided herein is admixed with a pharmaceutically acceptable carrier or excipient. See, e.g., remington's Pharmaceutical Sciences and U.S. Pharmacopeia: national Formulary, mack Publishing Company, easton, pa (1984).

Formulations of therapeutic and diagnostic agents may be prepared by mixing them with acceptable carriers, excipients or stabilizers in the form of, for example, lyophilized powders, slurries, aqueous solutions or suspensions (see, for example, hardman et al, (2001) Goodman and Gilman's The Pharmac ological Basis of Therapeutics, mcGraw-Hill, new York, N.Y., genn aro (2000) Remington: the Science and Practice of Pharmacy, lipp incatt, williams, and Wilkins, new York, N.Y., avis et al (eds.) (1993) Pharmaceutical Dosage Forms: parenteral Medications, marcel Dekker, N.Y., lieberman et al (1990) Pharmaceutical Dosage Forms:Tablets, marcel Dekker, N.Y., lieberman et al (1990) Pharmace utical Dosage Forms:Disperse Systems, marcel Dekker, N.Y., weine r and Intkoskie (2000) Excipient Toxicity and Safety, marcel Dekker, N.Y., ko.). In some embodiments, the antibody is diluted to a suitable concentration in a sodium acetate solution pH5 to 6 and NaCl or sucrose is added for tonicity. Additional agents such as polysorbate 20 or polysorbate 80 may be added to enhance stability.

Toxicity and therapeutic efficacy of antibody compositions administered alone or in combination with another dose can be determined in cell culture or experimental animals by standard pharmaceutical procedures, e.g., to determine LD ₅₀ (50% of the population lethal dose) and ED ₅₀ (50% of the population at a therapeutically effective dose). The dose ratio between toxic and therapeutic effects is the therapeutic index (LD ₅₀ /ED ₅₀ ). In particular aspects, antibodies exhibiting high therapeutic indices are desirable. The data obtained from these cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds is preferably within a range of circulating concentrations, including ED which is little or no toxic ₅₀ . The dosage may vary within this range depending upon the dosage form employed and the route of administration.

In some embodiments, the compositions of the invention are administered to a subject according to the Physics' Desk Reference 2003 (Thomson Healthcare; 57 th edition (11/1/2002)).

The mode of administration may vary. Suitable routes of administration include oral, rectal, transmucosal, enteral, parenteral; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, cutaneous, transdermal or intraarterial.

In some embodiments, the antibody or antigen binding fragment thereof may be administered by an invasive route such as injection. In some embodiments, the antibody or antigen-binding fragment thereof or pharmaceutical composition thereof is administered intravenously, subcutaneously, intramuscularly, intraarterially, intra-articular (e.g., in an arthritic joint), or by inhalation, aerosol delivery. Administration by a non-invasive route (e.g., oral; e.g., in a pill, capsule, or tablet) is also within the scope of embodiments of the present invention.

In some embodiments, the antibody or antigen binding fragment thereof may be administered directly to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retroorbital sinus. In some embodiments, the injection is into the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retroorbital Dou Shiyong. In some embodiments, the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, an suprachoroidal injection, or an intracameral injection. In some embodiments, the injection is intravitreal injection. In some embodiments, the injection is an intraorbital injection. In some embodiments, the injection is a retroorbital injection. In some embodiments, the injection is a suprachoroidal injection. In some embodiments, the injection is an intracameral injection.

In some embodiments, the anti-IGF-1R antibody or antigen-binding fragment thereof is administered in combination with at least one additional therapeutic agent, such as, but not limited to, any therapeutic agent used to treat thyroid-eye disease. For example, in some embodiments, an anti-IGF-1R antibody or antigen-binding fragment thereof is administered in combination with at least one other therapeutic agent, such as, but not limited to, a therapeutic agent to treat thyroid eye disease or a condition associated therewith. Examples of such therapeutic and therapeutic agents include, but are not limited to, antithyroid agents, diabetic agents, beta-blockers, propylthiouracil (propylthiouracil), methimazole (methimazole), propranolol, atenolol, metoprolol, nadolol, corticosteroids, metformin, sulfonylurea, meglitinide, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, conventional insulin, insulin aspart, insulin lispro, insulin protamine (insulin isophalne), deglupine, insulin diglucose, acarbose miglol (migllol), acebutolol (acebutolol), atenolol (atenolol), betaxolol (betaxolol), bisoprolol (bisoprolol), carteolol (cartelol), carvedilol (carvedilol), esmolol (esmolol), labetalol (labetalol), metoprolol (metalol), nadolol (nadolol), nebivolol (nebivolol), penbutolol (pentallolol), indolol (pindolol), propranolol (propranolol), sotalol (sotalol), timolol (timolol), timolol (tomolol) ophthalmic solution, sitagliptin (sitagliptin), saxagliptin, linagliptin (alotin), alogliptin (alogliptin), duraglutide, exenatide, semaglutin, liraglutide, canagliflozin, dapagliflozin, engagliflozin, or any combination thereof.

The composition may be administered by medical devices known in the art. For example, the pharmaceutical compositions of the present invention may be administered by injection with a hypodermic needle (including, for example, a pre-filled syringe or an auto-injector).

The pharmaceutical composition may also be administered with a needleless subcutaneous injection device; such as U.S. patent No. 6,620,135; no. 6,096,002; no. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; devices disclosed in 4,790,824 or 4,596,556.

The pharmaceutical composition may also be administered by infusion. Examples of well known implants and modular forms for administration of pharmaceutical compositions include: U.S. patent No. 4,487,603, which discloses an implantable micro-infusion pump that dispenses a drug at a controlled rate; U.S. Pat. No. 4,447,233, which discloses a drug infusion pump that delivers a drug at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses a variable flow implantable infusion device for continuous drug delivery; U.S. Pat. No. 4,439,196 discloses an osmotic drug delivery system having multiple compartment compartments. Many other such implants, delivery systems and modules are well known to those skilled in the art.

Alternatively, the antibody may be administered locally rather than systemically, for example by direct injection of the antibody into an arthritic joint or pathogen-induced lesion characterized by an immunopathology, typically in the form of a depot or sustained release formulation. Furthermore, the antibodies may be administered in the form of a targeted drug delivery system (e.g., in the form of liposomes coated with tissue specific antibodies) that targets, for example, arthritic joints or pathogen-induced lesions characterized by immune pathology. Liposomes will target and be selectively taken up by diseased tissue.

The administration regimen depends on several factors, including the serum or tissue turnover rate of the therapeutic antibody, the extent of the symptoms, the immunogenicity of the therapeutic antibody, and the accessibility of the target cells in the biological matrix. Preferably, the administration regimen delivers sufficient therapeutic antibody to effect amelioration of the disease condition of interest while minimizing undesirable side effects. Thus, the amount of biologic delivered depends in part on the particular therapeutic antibody and the severity of the condition being treated. Guidelines for selecting appropriate doses of therapeutic antibodies are available (see, e.g., wawrzynzak (1996) anti-body Therapy, bio Scientific Pub. Ltd, oxfordshire, UK; kresina (eds.) (1991) Monoclonal Antibodies, cytokines and Arthritis, marcel Dekker, new York, NY; bach (eds.) (1993) Monoclonal Antibodies and Peptide Therapy in Autoimmune Diseases, marcel Dekker, new York, NY; baert et al (2003) New Engl. J. Med.348:601-608; milgom et al (1999) New Engl. J. Med.341:1966-1973; slamon et al (2001) New Engl. J. Med.344:783-792; benenoviz et al (2000) New Engl. J. 613:613-342:2001-608; med. 46:46; new Engl. 46-46:46; med. 46:619).

The clinician, for example, determines the appropriate dosage using parameters or factors known or suspected in the art to affect the treatment. Generally, the initial dose is an amount slightly less than the optimal dose, followed by an incremental increase in smaller increments until the desired or optimal effect (relative to any negative side effects) is achieved. Important diagnostic measures include, for example, those measures of symptoms of inflammation or the level of inflammatory cytokines produced. In general, it is desirable that the biological agent to be used is derived from the same species as the targeted treated animal, thereby minimizing any immune response to the agent. In the case of human subjects, for example, chimeric, humanized and fully human antibodies may be desirable.

The antibody or antigen binding fragment thereof may be provided by continuous infusion or by a dose administered, for example, once daily, 1 to 7 times weekly, once weekly, biweekly, once monthly, bi-monthly, quarterly, half a year, once annually, etc. The dosage may be provided, for example, intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscularly, intracranially, intravertebrally, or by inhalation. In some embodiments, the antibody is administered every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks. In some embodiments, the antibody is administered every four weeks. In some embodiments, the antibody is administered every five weeks. In some embodiments, the antibody is administered every seven weeks. In some embodiments, the antibody is administered every six weeks. In some embodiments, the antibody is administered every eight weeks. In some embodiments, the antibody is administered for at least 21 to 52 weeks or longer. In some embodiments, the antibody is administered in this schedule for at least 21 weeks. In some embodiments, the antibody is administered at this schedule for at least 24 weeks. In some embodiments, the antibody is administered at this schedule for at least 32 weeks. In some embodiments, the antibody is administered at this schedule for at least 36 weeks. In some embodiments, the antibody is administered at this schedule for at least 40 weeks. In some embodiments, the antibody is administered at this schedule for at least 42 weeks. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) once. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) twice. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) three times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) four times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) five times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) six times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) seven times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) eight times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) nine times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 10 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 11 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 12 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 13 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 14 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 15 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 16 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 17 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 18 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 19 times. In some embodiments, the antibody is administered (e.g., infused or subcutaneously) 20 times. When an antibody is administered more than once, the antibody may be administered according to a schedule, such as the schedule provided herein.

Total weekly dosages may be provided as herein. In some embodiments, the total weekly dose is at least 0.05 μg/kg body weight, more generally at least 0.2 μg/kg, 0.5 μg/kg, 1 μg/kg, 10 μg/kg, 100 μg/kg, 0.25mg/kg, 1.0mg/kg, 2.0mg/kg, 5.0mg/ml, 10mg/kg, 25mg/kg, 50mg/kg or more (see, e.g., yang et al (2003) New Engl. J. Med.349:427-434; herld et al (2002) New Engl. J. Med.346:1692-1698; liu et al (1999) J. Neurol. Neurosurg. Psych.67:451-456; portielji et al (20003) Cancer Immunol. Immunother. 52:133-144). Dosages may also be provided to achieve a predetermined target concentration of antibodies in the serum of a subject, such as 0.1 μg/ml, 0.3 μg/ml, 1 μg/ml, 3 μg/ml, 10 μg/ml, 30 μg/ml, 100 μg/ml, 300 μg/ml, or greater than 300 μg/ml.

In some embodiments, the serum concentration of the antibody in the subject is at least or about 10 μg/ml or 20 μg/ml or 50 μg/ml, 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml or 105 μg/ml at least 1, 2 or 3 weeks after administration.

In some embodiments, a dose of 20mg/kg IV is administered. In some embodiments, one administration is used to provide a Cmin of 133ug/mL after about 5 weeks. In some embodiments, the administered antibody dose provides a Cmin of 102ug/mL after 6 weeks. In some embodiments, the dosage of antibody is as provided herein, e.g., 10mg/mg as the starting dose, with subsequent doses being the same or lower. In some embodiments, the antibody is administered as provided herein at a dose that achieves a Cmin of at least or about 100 ug/mL.

As used herein, "inhibiting" or "treating" includes delaying the progression of symptoms associated with a disorder and/or lessening the severity of symptoms of such a disorder. The term further includes ameliorating existing uncontrolled or unwanted symptoms, preventing additional symptoms and ameliorating or preventing the underlying cause of such symptoms. Thus, the term indicates that a beneficial result has been conferred to a vertebrate subject having, or likely to be afflicted with, a disorder, disease or condition.

As used herein, the terms "therapeutically effective amount," "therapeutically effective dose," and "effective amount" refer to an amount of an antibody or antigen-binding fragment thereof that is effective to cause a measurable improvement in one or more symptoms of a disease or condition or progression of such disease or condition when administered to a cell, tissue, or subject, alone or in combination with additional therapeutic agents. A therapeutically effective dose further refers to an amount of a binding compound sufficient to cause at least a partial improvement in symptoms, e.g., to treat, cure, prevent, or ameliorate a related medical condition, or to increase the rate of treatment, cure, prevention, or amelioration of such a condition. When applied to an individual active ingredient administered alone, a therapeutically effective dose refers to the ingredient alone. When applied to a combination, a therapeutically effective dose refers to the combined amount of the active ingredients that, whether administered in combination, serially or simultaneously, produces a therapeutic effect. An effective amount of the therapeutic agent will improve the diagnostic measure or parameter by at least 10%; typically at least 20%; preferably at least about 30%; more preferably at least 40%, most preferably at least 50%. In cases where subjective measures are used to assess disease severity, an effective amount may also result in an improvement in the subjective measures. In some embodiments, if the amount is an amount useful for treating or ameliorating a condition as provided herein, the amount is a therapeutically effective amount.

The term "subject" as used throughout includes any organism, such as animals, including mammals (e.g., rats, mice, dogs, cats, rabbits) and, for example, humans. The subject may also be referred to as a patient. In some embodiments, the subject is a subject in need thereof. A subject "in need" refers to a subject that has been identified as in need of treatment for a condition to be treated and is treated with a particular intent to treat such condition. For example, the condition may be any of the conditions described herein.

In view of the isolated antibodies binding to epitopes on IGF-1R proteins or other proteins described herein and exhibiting IGF-1R inhibiting or therapeutic activity in vitro and/or in vivo, antibodies or antigen binding fragments thereof capable of inhibiting IGF-1R function are suitable for use as therapeutic agents for the treatment of IGF-1R related conditions in humans and animals. These conditions include thyroid eye disease. Thus, also provided are methods of treating such conditions, wherein the methods comprise administering an antibody or antigen-binding fragment thereof to a subject having such a condition.

In some embodiments, the method comprises administering to a susceptible subject or a subject exhibiting a condition in which IGF-1R is known or suspected to have caused the observed pathology a therapeutically or prophylactically effective amount of one or more monoclonal antibodies or antigen binding fragments of an antibody described herein. Any active form of antibody may be administered, including, but not limited to, scFv, fab, and F (ab') 2 fragments, and other forms of antibodies provided herein.

As used herein, IGF-1R-related pathology refers to a pathology caused by modulation of IGF-1R. These conditions include, but are not limited to, thyroid eye disease and other conditions provided herein.

In some embodiments, the antibodies used are compatible with the recipient species such that the immune response against the MAb does not produce an unacceptably short circulation half-life or induce an immune response against the MAb in the subject.

Treatment of an individual may include administering a therapeutically effective amount of an antibody described herein. Antibodies may be provided in kit form, such as those provided herein. The antibody may be used or administered alone or in admixture with another therapeutic, analgesic or diagnostic agent as provided herein. In providing an antibody or fragment thereof capable of binding to IGF-1R or an antibody capable of preventing IGF-1R pathology in a recipient patient to the patient, the dosage of the administered agent will vary depending on factors such as the age, weight, height, sex, general medical condition, past medical history, etc. of the patient.

An antibody capable of treating a condition associated with IGF-1R activity or to treat an IGF-1R related pathology is intended to be provided to a subject in an amount sufficient to affect a reduction, regression, or improvement in IGF-1R related symptoms or pathology. Such pathologies include thyroid eye diseases and the like.

Thus, in some embodiments, methods of treating a subject having an IGF-1R mediated disorder are provided. In some embodiments, the methods comprise administering a pharmaceutical composition comprising an antibody or antigen-binding fragment thereof as provided herein. In some embodiments, the disorder is thyroid eye disease. As provided herein, the antibodies or antigen binding fragments thereof may be administered with other therapeutic agents. These therapeutic agents may be administered simultaneously or sequentially.

In some embodiments, the antibodies or antigen binding fragments thereof are useful for treating thyroid eye disease. In some embodiments, the antibodies or antigen binding fragments thereof are useful for treating or lessening the severity of thyroid-related eye disease (TAO) or symptoms thereof.

In some embodiments, methods or uses are provided to reduce the ocular bulge in a subject's eye suffering from thyroid-related eye disease (TAO).

In some embodiments, the subject is a subject that has been previously treated with antibodies different from those provided herein.

In some embodiments, methods or uses are provided for Clinical Activity Scoring (CAS) of a subject having or suspected of having a thyroid-related eye disease (TAO).

In some embodiments, methods or uses are provided for reducing the relief of the eye of a subject suffering from thyroid-related eye disease (TAO) by at least 2mm and b) reducing Clinical Activity Score (CAS).

As used herein, the term Clinical Activity Scoring (CAS) refers to a scheme described and scored according to table 2. According to this scheme, there is a score given for each parameter evaluated in the table below. The sum of all scores defines clinical activity and provides CAS, with 0 or 1 constituting the inactive disease and 7 being the severely active ocular disease.

As provided in table 2, CAS is composed of seven components: spontaneous retrobulbar pain, pain in attempts to move the eye (up, left, right, and down gaze), conjunctival redness, palpebral redness, bulbar conjunctival edema, verruca/fold swelling, and eyelid swelling. The components were scored for the presence (1 point) or absence (0 point). The score for each efficacy assessment is the sum of all present items; a range of 0 to 7 is obtained, where 0 or 1 constitutes an inactive disease and 7 is a severely active ocular disease. A >2 score change is considered clinically significant.

The method of claim 1, wherein the spontaneous orbital pain is pain sensation or pressure sensation on or behind the sphere. This pain can be caused by elevated intra-orbital pressure as the volume of orbital tissue increases through excessive synthesis of extracellular matrix, fluid accumulation, and cellular infiltration and expansion. The gaze-induced orbital pain of item 2 may be eye pain when looking or attempting to look up, down or sideways, i.e. pain when the eye moves up, down or sideways or when attempting to move the eye. Such pain may be caused by inflamed muscle stretching, especially when attempting to look upward. The 'stretching pain' is not caused by the finger pressing on the eyeball and would be predicted if it were a manifestation of elevated intra-orbital pressure. Both types of pain may be reduced after anti-inflammatory treatment. Thus, these types of pain are believed to be directly associated with autoimmune inflammation of the orbit and are therefore suitable for assessing TAO activity.

Swelling in TAO was considered as term 6 bulbar conjunctiva edema (conjunctival edema) in table 1, and verruca and/or semilunar fold swelling. Both are signs of TAO activity. Eyelid swelling can be caused by edema, fat prolapse of the entire orbital membrane, or fibrosis. In addition to swelling, other symptoms indicative of active TAO include redness and/or pain of the conjunctiva, eyelid, mons and/or folds of the semilunar fold.

In some embodiments, the subject's eyes are reduced by at least 2mm. In some embodiments, the subject's eyes are reduced by at least 3mm. In some embodiments, the subject's eyes are reduced by at least 4mm.

In some embodiments, in the subject being treated, the subject's Clinical Activity Score (CAS) is reduced by at least 2 points. In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to one (1). In some embodiments, the subject's Clinical Activity Score (CAS) decreases to zero (0).

In some embodiments, there is provided a method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject, wherein treatment with said antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

In some embodiments, methods of improving the quality of life of a subject suffering from thyroid-related eye disease (TAO, also known as graves 'eye disease/graves' orbit disease) are provided. In some embodiments, quality of life is measured by graves' eye disease quality of life (GO-QoL) assessment or its vision function or appearance subscale. In some embodiments, the treatment results in a GO-QoL improvement of greater than or equal to 8 points. In some embodiments, the treatment results in an improvement in the GO-QoL functional sub-scale. In some embodiments, the treatment results in an improvement in the appearance sub-scale of GO-QoL.

In some embodiments, methods of treating or lessening the severity of a double vision in a subject suffering from thyroid-related eye disease (TAO) are provided. In some embodiments, the complex is considered a constant complex. In some embodiments, the review is a non-constant review. In some embodiments, the review is considered intermittent review. In some embodiments, improvement in or reduction in severity of the double vision persists for at least 20 weeks after cessation of antibody administration. In some embodiments, improvement in or reduction in severity of the double vision persists for at least 50 weeks after cessation of antibody administration.

The severity of the disease can be measured in the following non-limiting embodiments. For example, for eyelid orifices, the distance (in mm) between the eyelid edges is measured while the patient is relaxed in the first eye position, sitting position, and looking at a distance. For swelling of the eyelid, measured/assessed as "absent/undefined", "moderate" or "severe". Eyelid redness is absent or present. Conjunctival redness is absent or present. In some embodiments, conjunctival edema is absent or present. In some embodiments, the inflammation of the verruca or plication is absent or present. The eyes were measured in millimeters using the same Hertel eye relief meter and the same angular separation for individual patients. Subjective review scores of 0 to 3 (0 = no review; 1 = review in the first gaze position at tiredness or at first wake-up; 2 = review at non-constant, i.e. extreme review; 3 = review at constant, i.e. continuous review in the first eye position or reading position). For ocular muscular involvement, monocular movement (reduction) is measured in degrees. Cornea involvement is absent/punctiform or keratopathy/ulceration. For optic nerve involvement, i.e., optimal correction of vision, color vision, optic disc, relative pupil afferent defects, the condition is absent or present. In addition, when optic nerve compression is suspected, the visual field is detected. In some embodiments, patients may be classified according to the following severity categories. For example, thyroid eye disease that endangers vision: patients with thyroid Dysfunction Optic Neuropathy (DON) and/or cornea rupture. Such category requires immediate intervention. Moderate to severe thyroid eye disease: patients without vision-compromised disease, but whose eye disease is sufficient to affect daily life, justify the risk of immunosuppression (if active) or surgical intervention (if inactive). Patients with moderate to severe thyroid eye disease typically have any one or more of the following: eyelid retraction is greater than or equal to 2mm, moderate or severe soft tissue involvement, and the protruding eye is 3mm above or 3mm above normal for race and sex, non-constant or constant double vision. Mild thyroid eye disease: the characteristics of thyroid eye disease have only a slight impact on daily life, and are insufficient to justify immunosuppression or surgical treatment in patients. They typically have only one or more of the following: slight eyelid withdrawal (< 2 mm), mild soft tissue involvement, raised eyes <3mm above normal values for race and sex, temporary or no compound vision, and corneal exposure in response to lubricant.

In some embodiments, the patient may be characterized by a graves' quality of life (GO-QoL) score. In addition to the bulge (or eye-pop) and CAS, GO quality of life (GO-QoL) questionnaires are used to evaluate quality of life. This questionnaire was designed to determine the improved quality of life following treatment using the methods disclosed herein. In some embodiments, the questionnaire may determine that the side effects after treatment with an antibody or antigen binding fragment thereof according to the methods disclosed herein are reduced or absent from the side effects when treated with a glucocorticoid. GO-QoL is a 16-item self-filling questionnaire divided into 2 subsets and used to evaluate the perceived impact of a subject's TED on (i) his daily physical activity associated with vision function and (ii) psychosocial function. Quality of life was assessed using a GO QoL questionnaire. The GO-QoL questionnaire was completed on days 1 and 6, 12 and 24 (or PW) and on months 7 and 12 (or PW) during the follow-up period [ C.B.Terwee et al, 1998]. GO-Qol is a 16 self-filling questionnaire divided into two self-evaluation sub-scales; one sub-scale covers the effect of vision function on daily activities and the other sub-scale evaluates the effect of self-perceived appearance. Vision function subscales cover activities such as driving, walking outdoors, reading, watching tv. The appearance sub-scale interrogates the subject for the following questions: such as whether an eye disease changes the appearance of the subject, causing others to have a negative response to the subject, causing social separation and causing the subject to attempt to mask its appearance. Each sub-table has 8 questions, so-this is very true; yes- -one point; or no-neither will answer the question. Each question was scored from 0 to 2 separately and then the total raw score was mathematically converted to a 0-100 scale, where 0 represents the greatest negative impact on quality of life and 100 represents no impact. Changes > or equal to 8 points on the 0-100 scale are shown to be clinically significant. The combined scores take the original scores from both sub-scales and convert them back to a single 0-100 scale. The questionnaire has two self-evaluating sub-scales. Each sub-table has 8 questions, to (i) be-very so; (ii) is-a point; or (iii) no-neither answer the question. Each question was scored from 0 to 2 separately, and then the total raw score was mathematically converted to a 0-100 scale, where 0 represents the greatest negative impact on quality of life and 100 represents no impact. A >8 point change on the 0-100 scale is considered clinically significant. The combined scores were taken from the original scores from both sub-scales and then converted to a single 0-100 scale.

Patients may also be assessed by the presence or absence of a review Gorman scale. The Gorman assessment of subjective review includes four categories: no compound vision (absence), compound vision when the patient is tired or awake (intermittent), compound vision when extremely focused (non-constant), and continuous compound vision in the first eye position or reading position (constant). Patients were scored according to their grade of review they experienced. Improvements of grade 1 or greater are considered clinically significant.

In some embodiments, the method comprises administering an antibody, such as those provided herein. In some embodiments, the antibody is administered as a first dose at a dose of about 1mg/kg to about 5mg/kg of antibody. In some embodiments, the antibody is administered as a first dose at a dose of about 5mg/kg to about 10mg/kg of antibody. In some embodiments, the antibody is administered as a subsequent dose at a dose of about 5mg/kg to about 20mg/kg of antibody. In some embodiments, the antibody is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses. In some embodiments, the subsequent dose is administered every three weeks for at least 21 weeks.

In some embodiments, the antibody is administered in the form of a pharmaceutical composition, such as those provided herein. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for treating TAO. In some embodiments, the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD 20 antibody; tobrazumab or other anti-IL-6 antibodies; or selenium, infliximab or other anti-tnfα antibodies or Thyroid Stimulating Hormone Receptor (TSHR) inhibitors.

In some embodiments, the methods provided herein comprise administering to a subject an antibody or antigen-binding fragment thereof that specifically binds and inhibits IGF-IR. In some embodiments, the antibodies are as provided herein.

Kits useful in practicing embodiments described herein are also provided. The kit of the invention comprises a first container containing or packaged in association with an antibody as described above. The kit may also comprise another container containing or packaged in association with a solution necessary or appropriate for the implementation. The container may be made of glass, plastic or foil and may be a vial, bottle, pouch, tube, bag, etc. Kits may also contain written information, such as the procedure used to implement the embodiments; or analytical information such as the amount of reagent contained in the first container means. The container may be located in another container apparatus (e.g., a box or bag) along with the written information.

Yet another aspect provided herein is a kit for detecting IGF-1R protein in a biological sample. The kit includes a container containing one or more antibodies that bind to epitopes of IGF-1R protein and instructions for using the antibodies for the following purposes: binding the antibody to the IGF-1R protein to form an immune complex and detecting the formation of the immune complex such that the presence or absence of the immune complex correlates with the presence or absence of the IGF-1R protein in the sample. Examples of containers include multi-well culture plates that allow for simultaneous detection of IGF-1R proteins in multiple samples.

In some embodiments, antibodies that bind to IGF-1R proteins are provided. In some embodiments, the antibody is isolated. In some embodiments, the antibody specifically binds. In some embodiments, the antibody binds to an appropriately folded IGF-1R protein. In some embodiments, the antibody is specific for a particular IGF-1R conformational state (open or closed). In some embodiments, the antibody binds to IGF-1R protein in the cell membrane. In some embodiments, the antibody binds to IGF-1R protein in the cell membrane in intact cells. In some embodiments, the antibody inhibits or neutralizes the function of IGF-1R protein. As used herein, the term "neutralize" means that the activity or function of a protein is inhibited. Inhibition may be complete or partial. In some embodiments, the activity or function of the protein is inhibited by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99%. The percent inhibition may be based on the function or activity of the protein in the absence of antibodies. In some embodiments, the antibody inhibits glucose transport facilitated by IGF-1R. In some embodiments, the antibody inhibits internalization of IGF-1R protein.

In some embodiments, the antibody comprises a sequence as provided herein or an antigen binding fragment thereof. In some embodiments, the antibody comprises a heavy chain CDR described herein or an antigen binding fragment thereof. The heavy chain may be one or more of the heavy chains described herein. In some embodiments, the antibody comprises a light chain or antigen-binding fragment thereof as described herein.

In some embodiments, methods of treating, inhibiting, or ameliorating an IGF-1R related pathology are provided. In some embodiments, the methods comprise administering an antibody described herein or a pharmaceutical composition described herein to a subject to treat, inhibit, or ameliorate IGF-1R-related pathology. In some embodiments, the pathology is as described herein.

In some embodiments, methods of detecting the presence or absence of IGF-1R in a sample are provided, the methods comprising contacting the sample with one or more antibodies described herein, thereby detecting binding of the antibodies to IGF-1R antigen. In some embodiments, detection of binding indicates the presence of IGF-1R antigen; or an absence of detection of binding to IGF-1R antigen indicates the absence of IGF-1R antigen. The detection may be performed by any known method, such as using a biosensor, ELISA, sandwich assay, etc. However, in some embodiments, the method includes detecting the presence of the protein in non-denaturing conditions. Non-denaturing conditions may be used so that the protein of interest is detected in its native or properly folded form.

In some embodiments, a method of identifying a test antibody that binds to an epitope on an IGF-1R protein is provided, the method comprising contacting the test antibody with the epitope on the IGF-1R protein and determining whether the test antibody binds to the epitope. In some embodiments, determining comprises determining whether the test antibody binds to a protein and is competitively inhibited by an antibody comprising a sequence as provided herein. In some embodiments, determining comprises mutating one or more residues of an epitope or protein and determining binding of the test antibody to the mutated epitope, wherein the test antibody is considered to bind to that epitope if the mutation reduces binding of the test antibody compared to a non-mutated epitope.

In some embodiments, methods of monitoring internalization of IGF-1R from the cell surface are provided. In some embodiments, the methods comprise contacting a cell with an anti-IGF-1R antibody as provided herein and detecting the presence of IGF-1R in or on the cell surface. Differences in cell surface expression can be measured and internalization can be monitored and measured. For example, this can be used to measure the effect of another molecule (e.g., a test agent) on regulating internalization of IGF-1R protein. Thus, the antibodies provided herein can be used to identify assays that modulate (increase or decrease) internalization of IGF-1R proteins. Test molecules that increase internalization, which would be measured as decreased binding of an anti-IGF-1R antibody to an IGF-1R protein on the cell surface, can be identified according to the methods provided herein. Test molecules that reduce internalization, which would be measured as increased binding of an anti-IGF-1R antibody to an IGF-1R protein on the cell surface, can be identified according to the methods provided herein. Surface expression may be measured by fluorescence, which may be performed by a secondary antibody recognizing IGF-1R antibodies or by labeling anti-IGF-1R antibodies provided herein.

In some embodiments, methods of inhibiting IGF-1 stimulated receptor phosphorylation on a cell are provided. In some embodiments, the method comprises contacting the cell with an antibody or pharmaceutical composition comprising the same as provided herein. In some embodiments, contacting comprises administering an antibody or a pharmaceutical composition comprising the same to a subject. In some embodiments, the cell is a cell in the eye. In some embodiments, the subject has or is at risk of Thyroid Eye Disease (TED). In some embodiments, the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm. In some embodiments, the IC50 is measured in an in vitro assay, as described in the examples, as provided herein. In some embodiments, the IC50 is measured in a cell that is an a549 cell or a HOCF cell.

In some embodiments, there is provided a method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml, or 105 μg/ml for at least 1, 2, or 3 weeks after administration. In some embodiments, the serum concentration is measured after administering one, two, or three doses of the antibody or pharmaceutical composition comprising the same to the subject.

In some embodiments, methods of inhibiting IGF-1 induced receptor autophosphorylation in a subject in need thereof by at least 95%, 96%, 97%, 98%, or 99% or 100% are provided. In some embodiments, the method comprises administering an antibody, or pharmaceutical composition comprising the same, as provided herein to a subject. In some embodiments, IGF-1 induced receptor autophosphorylation is inhibited in the eye or orbital region of a subject. In some embodiments, IGF-1 induced receptor autophosphorylation is inhibited, thereby treating thyroid eye disease or ameliorating symptoms as described herein in a subject.

The illustrated embodiments

In some embodiments, embodiments provided herein further include (but are not limited to):

1. an antibody or antigen-binding fragment thereof, comprising:

a VL sequence as set forth in SEQ ID NOs 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86;

VH sequences as set forth in SEQ ID NOs 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83;

an LCDR sequence as set forth in SEQ ID NO 17, 18, 19, 23, 24, 25, 29, 30, 31, 35, 36, 37, 41, 42, 43, 47, 48, 49, 53, 54, 55, 59, 60, 61 or 81, or

An HCDR sequence as set forth in SEQ ID NO 20, 21, 22, 26, 27, 28, 32, 33, 34, 38, 39, 40, 44, 45, 46, 50, 51, 52, 56, 57, 58, 62, 63 or 64; and

Any combination or variant thereof.

2. The antibody or antigen-binding fragment thereof of embodiment 1, wherein the antibody binds to IGF-1R.

3. The antibody of embodiment 1, wherein the antibody is a monoclonal antibody.

4. The antibody of embodiment 1, wherein the antibody is a humanized antibody.

5. The antibody of embodiment 1, wherein the antibody is an scFv antibody.

6. The antibody of any one of embodiments 1 to 5, wherein the antibody or antigen-binding fragment thereof comprises V as set forth in SEQ ID NOs 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86 _L A peptide, or any variant thereof.

7. The antibody of any one of embodiments 1 to 6, wherein the antibody or antigen-binding fragment thereof comprises V as set forth in SEQ ID NO 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83 _H A peptide, or any variant thereof.

8. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO:20, 26, 32, 38, 44, 50 or 56; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO. 21, 27, 33, 39, 45, 51 or 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 22, 28, 34, 40, 46, 52 or 58; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 17, 23, 29, 35, 41, 47 or 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 18, 24, 30, 36, 42, 48 or 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 19, 25, 31, 37, 43, 49, 55 or 81; or a variant of any of the foregoing.

9. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 20; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 21; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 22; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 17; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 18; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 19; or a variant of any of the foregoing.

10. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 26; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 27; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 28; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 23; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 24; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 25; or a variant of any of the foregoing.

11. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 32; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 33; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 34; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 29; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 30; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 31; or a variant of any of the foregoing.

12. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 37; or a variant of any of the foregoing.

13. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 44; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 45; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 46; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 41; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 42; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 43; or a variant of any of the foregoing.

14. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 50; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 51; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 52; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 47; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 48; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 49; or a variant of any of the foregoing.

15. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 56; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 57; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 58; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 53; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 54; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 55; or a variant of any of the foregoing.

16. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 62; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 63; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 64; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 59; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 60; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 61; or a variant of any of the foregoing.

17. An antibody or antigen-binding fragment thereof, wherein the antibody or antibody fragment comprises: (i) A heavy chain variable region comprising heavy chain CDR1, CDR2 and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID No. 38; the heavy chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 39; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2 and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID No. 35; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO. 36; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO. 81; or a variant of any of the foregoing.

18. The antibody of any one of embodiments 6-17, wherein the heavy chain variable region and the light chain variable region are not linked by a linker.

19. The antibody of any one of embodiments 6-17, wherein the heavy chain variable region and the light chain variable region are linked by a peptide linker.

20. The antibody of embodiment 19, wherein the peptide linker comprises the sequence of: (GGGGS) _n (SEQ ID NO:73)；(GGGGA) _n (SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5.

21. The antibody of any one of embodiments 1 to 20, wherein the antibody comprises the sequence of SEQ ID NOs 65 to 72, 78, 82 or 85, or variants thereof.

22. The antibody of any one of embodiments 1 to 21, wherein the antibody comprises V as set forth herein _L Sequence: 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86, or a variant thereof.

23. The antibody of any one of embodiments 1 to 21, wherein the antibody comprises V as set forth in SEQ ID NO 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83 _H A sequence, or a variant thereof.

24. The isolated antibody of any one of embodiments 1 to 21, wherein the antibody comprises the sequence of SEQ ID NOs 65 to 72, 78, 82 or 85, or variants thereof.

25. The antibody of any one of embodiments 1 to 24, wherein the variant has 1 to 10 substitutions, deletions or insertions.

26. The antibody of any one of embodiments 1 to 24, wherein the variant has 1 to 10 conservative substitutions.

27. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 85% homology to the sequence of SEQ ID NOs 1 to 72, 78 to 83, or 85 to 86.

28. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homology to the sequence of SEQ ID NOs 1 to 72, 78 to 83 or 85 to 86.

29. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 85% identity to the sequence of SEQ ID NOs 1 to 72, 78 to 83 or 85 to 86.

30. The antibody of any one of embodiments 1 to 26, wherein the variant has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the sequence of SEQ ID NOs 1 to 72, 78 to 83 or 85 to 86.

31. The antibody of any one of embodiments 1-26, wherein the antibody is an scFv antibody.

32. The antibody of any one of embodiments 1 to 26, wherein the antibody is a monoclonal antibody.

33. The antibody of any one of embodiments 1 to 26, wherein the antibody is a humanized antibody.

34. The antibody of any one of the preceding embodiments, wherein the antibody comprises an Fc region.

35. The antibody of embodiment 34, wherein the Fc region is as set forth in SEQ ID NOS 75 to 77 or 84.

36. The antibody of any one of the preceding embodiments, wherein the Fc region comprises a mutation that when linked to the Fc region increases the half-life of the antibody.

37. The antibody of embodiment 36, wherein the Fc region comprises the S228P, L235E, M Y, S254T, T256E, M428L, N434S, L35234F, P S mutation or any combination thereof.

38. The antibody of embodiment 36, wherein the Fc region comprises M252Y, S254T and T256E mutations.

39. The antibody of embodiment 36, wherein the Fc region comprises S228P and L235E mutations.

40. The antibody of embodiment 36, wherein the Fc region comprises L234F, L235E and P331S mutations.

41. The antibody of embodiment 36, wherein the Fc region comprises M252Y, S254T, T256E, S228P and L235E mutations.

42. The antibody of embodiment 36, wherein the Fc region comprises S228P, L235E, M L and N434S mutations.

43. The antibody of embodiment 36, wherein the Fc region comprises M428L and N434S mutations.

44. The antibody of embodiment 36, wherein the Fc region comprises L234F, L235E, P331S, M Y, S254T and T256E mutations.

45. A nucleic acid molecule encoding the antibody or antigen-binding fragment thereof of any one of the preceding embodiments.

46. A vector comprising the nucleic acid molecule of embodiment 45.

47. A cell comprising a nucleic acid molecule according to embodiment 45 or a vector according to embodiment 46.

48. A pharmaceutical composition comprising the antibody or nucleic acid molecule encoding the same of any one of embodiments 1-44.

49. The pharmaceutical composition of embodiment 48, wherein the composition is an injectable pharmaceutical composition.

50. A method of treating or lessening the severity of thyroid-associated eye disease (TAO) or a symptom thereof, comprising administering to a subject the antibody of any one of embodiments 1-44 or a pharmaceutical composition comprising the same.

51. A method of reducing the ocular bulge in a subject suffering from thyroid-associated ocular disease (TAO), comprising administering to the subject the antibody of any one of embodiments 1-44 or a pharmaceutical composition comprising the same.

52. A method of treating thyroid eye disease in a subject comprising administering to the subject the antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising the same.

53. A method of reducing the Clinical Activity Score (CAS) of a thyroid-associated eye disease (TAO) in a subject comprising administering to the subject the antibody of any one of embodiments 1-44, or a pharmaceutical composition comprising the same.

54. A method of reducing a) the bulge eye by at least 2mm and b) the Clinical Activity Score (CAS) in a subject having a thyroid-associated eye disease (TAO) comprising administering to the subject the antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising the same.

55. The method of any one of embodiments 50 to 54, wherein the protrusion is reduced by at least 2mm.

56. The method of any one of embodiments 50 to 54, wherein the protrusion is reduced by at least 3mm.

57. The method of any one of embodiments 50 to 54, wherein the protrusion is reduced by at least 4mm.

58. The method of any one of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

59. The method of any one of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced to one (1).

60. The method of any one of embodiments 50-54, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

61. A method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject comprising administering to a subject the antibody of any one of embodiments 1-44, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

62. A method of improving the quality of life of a subject suffering from thyroid-associated ocular disease (TAO, also known as graves 'eye disease/graves' orbit disease) comprising administering to the subject the antibody of any one of embodiments 1 to 44, or a pharmaceutical composition comprising the same.

63. The method of embodiment 62, wherein quality of life is measured by a graves' eye disease quality of life (GO-QoL) assessment or a visual function or appearance subscale thereof.

64. The method of embodiment 63, wherein the treatment results in a GO-QoL improvement of greater than or equal to 8 points.

65. The method of embodiment 63, wherein the treatment results in an improvement in the GO-QoL functional sub-scale.

66. The method of embodiment 63, wherein the treatment results in an improvement in the appearance subscale of GO-QoL.

67. A method of treating or lessening the severity of a double vision in a subject having a thyroid-associated eye disease (TAO), comprising administering to the subject the antibody of any one of embodiments 1-44, or a pharmaceutical composition comprising the same.

68. The method of embodiment 67, wherein the review is a constancy review.

69. The method of embodiment 67, wherein the review is non-constant review.

70. The method of embodiment 67, wherein the review is considered intermittent review.

71. The method of embodiment 67, wherein the improvement in or the severity of the double vision is reduced for at least 20 weeks after cessation of antibody administration.

72. The method of embodiment 67, wherein the improvement in or the severity of the double vision is reduced for at least 50 weeks after cessation of antibody administration.

73. The method of any one of embodiments 50 to 72, wherein the antibody is administered as a first dose at a dose of about 1mg/kg to about 5mg/kg antibody.

74. The method of any one of embodiments 50 to 72, wherein the antibody is administered as a first dose at a dose of about 5mg/kg to about 10mg/kg of antibody.

75. The method of any one of embodiments 50 to 72, wherein the antibody is administered as a subsequent dose at a dose of about 5mg/kg to about 20mg/kg antibody.

76. The method of any one of embodiments 50 to 72, wherein the antibody is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses.

77. The method of embodiment 76, wherein the subsequent dose is administered every three weeks for at least 21 weeks.

78. The method of any one of embodiments 50 to 77, wherein the antibody or antigen-binding fragment thereof is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a bispecific antibody, a single chain antibody, a multispecific antibody, a Fab ', a F (ab') 2, an Fv, or a scFv.

79. The method of any one of embodiments 50 to 78, wherein the antibody or antigen-binding fragment thereof is administered in the form of a pharmaceutical composition further comprising a pharmaceutically acceptable diluent or excipient or carrier.

80. The method of embodiment 79, wherein the pharmaceutical composition further comprises one or more pharmaceutically active compounds for treating TAO.

81. The method of embodiment 79 or 80, wherein the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD 20 antibody; tobrazumab or other anti-IL-6 antibodies; or selenium, infliximab or other anti-tnfα antibodies or Thyroid Stimulating Hormone Receptor (TSHR) inhibitors.

82. The method of any one of the preceding embodiments, wherein the antibody or antigen binding fragment thereof is administered directly to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retroorbital sinus.

83. The method of embodiment 82, wherein the antibody or antigen-binding fragment thereof is administered by injection.

84. The method of embodiment 83, wherein the injection is intravitreal, intraorbital, retroorbital, suprachoroidal or intracameral.

85. A method of increasing IGF-1R internalization on a cell, the method comprising contacting the cell with the antibody of any one of embodiments 1 to 44 or a pharmaceutical composition comprising the same.

86. The method of embodiment 85, wherein contacting comprises administering to the subject the antibody of any one of embodiments 1 to 44 or a pharmaceutical composition comprising the same.

87. The method of embodiment 86, wherein the subject has or is at risk of thyroid-eye disease (TED).

88. A method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with the antibody of any one of embodiments 1-44 or a pharmaceutical composition comprising the same.

89. The method of embodiment 88, wherein contacting comprises administering to a subject the antibody of any one of embodiments 1-44 or a pharmaceutical composition comprising the same.

90. The method of embodiment 89, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

91. The method of any one of embodiments 88 to 90, wherein the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm.

92. The method of embodiment 91, wherein the IC50 is measured in an in vitro assay, as provided herein.

93. The method of any one of embodiments 88 to 92, wherein the cell is an a549 cell or an HOCF cell.

94. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody of any one of embodiments 1 to 44 or as provided elsewhere herein, or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml, or 105 μg/ml for at least 1, 2, or 3 weeks after administration.

95. The method of embodiment 94, wherein the antibody or pharmaceutical composition is administered intravenously.

96. The method of embodiment 94 or 96, wherein the antibody or pharmaceutical composition is administered at a dose of about 20 mg/kg.

97. The method of any one of embodiments 94 to 96, wherein the antibody or pharmaceutical composition is administered at least or about once a week, once every two weeks, once every 3 weeks, or once every 4 weeks.

98. A method of inhibiting IGF-1 induced receptor autophosphorylation in a cell by at least 95%, 96%, 97%, 98% or 99% or 100%, comprising contacting the cell with an antibody or pharmaceutical composition comprising the same as described in any one of embodiments 1 to 44 or as provided elsewhere herein.

99. The method of embodiment 98, wherein inhibition of IGF-1 induced receptor autophosphorylation is measured as compared to receptor autophosphorylation induced in the absence of the antibody or pharmaceutical composition.

100. The method of embodiment 98 or 99, wherein contacting comprises administering an antibody or a pharmaceutical composition comprising the same to the subject.

101. The method of embodiment 100, wherein the subject has or is at risk of thyroid-eye disease (TED).

102. A method of inhibiting IGF-1 induced receptor autophosphorylation in a subject in need thereof by at least 95%, 96%, 97%, 98% or 99% or 100%, comprising administering to the subject an antibody of any one of embodiments 1 to 44 or as provided elsewhere herein or a pharmaceutical composition comprising the same.

103. The method of embodiment 102, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

104. The method of any one of embodiments 102 or 103, wherein the antibody or pharmaceutical composition is administered intravenously.

105. The method of any one of embodiments 98 to 104, wherein the antibody comprises a CDR of VRDN-1100.

106. The method of any one of embodiments 98 to 104, wherein the antibody comprises a CDR of an antibody to VRDN-1100 or a CDR of VRDN-2700.

107. An isolated antibody comprising a light chain having the amino acid sequence of SEQ ID No. 3 and a heavy chain comprising the amino acid sequence of SEQ ID No. 83.

108. An isolated antibody comprising a light chain variable region having the amino acid sequence of SEQ ID No. 13 and a heavy chain variable region having the amino acid sequence of SEQ ID No. 14.

109. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain amino acid sequence of SEQ ID NO. 92.

110. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain amino acid sequence of SEQ ID NO. 94.

111. The isolated antibody of embodiment 108, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID NO. 93 and a heavy chain having the amino acid sequence of SEQ ID NO. 95.

112. A pharmaceutical composition comprising the antibody of any one of embodiments 107-111.

113. A pharmaceutical composition suitable for intravenous administration comprising the antibody of any one of embodiments 107-111.

114. A pharmaceutical composition suitable for subcutaneous administration comprising the antibody of any one of embodiments 107-111.

115. A method of treating thyroid eye disease in a subject, the method comprising administering a pharmaceutical composition comprising the antibody of any one of embodiments 107-111.

116. The method of embodiment 115, wherein the pharmaceutical composition is administered intravenously.

117. The method of embodiment 115, wherein the pharmaceutical composition is administered subcutaneously.

118. A method of treating or lessening the severity of thyroid-associated eye disease (TAO) or a symptom thereof, comprising administering to a subject the antibody of any one of embodiments 107-111 or a pharmaceutical composition comprising the same.

119. A method of reducing the ocular bulge in a subject suffering from thyroid-associated ocular disease (TAO), comprising administering to the subject the antibody of any one of embodiments 107-111 or a pharmaceutical composition comprising the same.

120. A method of treating thyroid eye disease in a subject comprising administering to the subject the antibody of any one of embodiments 107-111 or a pharmaceutical composition comprising the same.

121. A method of reducing the Clinical Activity Score (CAS) of a thyroid-associated eye disease (TAO) in a subject, comprising administering to the subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same.

122. A method of reducing a) the bulge eye by at least 2mm and b) the Clinical Activity Score (CAS) in a subject having a thyroid-associated eye disease (TAO) comprising administering to the subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same.

123. The method of any one of embodiments 118 to 122, wherein the protrusion is reduced by at least 2mm.

124. The method of any one of embodiments 118 to 122, wherein the protrusion is reduced by at least 3mm.

125. The method of any one of embodiments 118 to 122, wherein the protrusion is reduced by at least 4mm.

126. The method of any one of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

127. The method of any one of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced to one (1).

128. The method of any one of embodiments 118-122, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

129. A method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject comprising administering to a subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

130. A method of improving the quality of life of a subject suffering from thyroid-associated ocular disease (TAO, also known as graves 'eye disease/graves' orbit disease), comprising administering to the subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same.

131. The method of embodiment 130, wherein quality of life is measured by a graves' eye disease quality of life (GO-QoL) assessment or a visual function or appearance subscale thereof.

132. The method of embodiment 130, wherein the treatment results in a GO-QoL improvement of greater than or equal to 8 points.

133. The method of embodiment 130, wherein the treatment results in an improvement in the GO-QoL function sub-scale.

134. The method of embodiment 130, wherein the treatment results in an improvement in the appearance sub-scale of GO-QoL.

135. A method of treating or lessening the severity of a double vision in a subject having a thyroid-associated eye disease (TAO), comprising administering to the subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same.

136. The method of embodiment 135, wherein the review is a constancy review.

137. The method of embodiment 135, wherein the review is non-constant review.

138. The method of embodiment 135, wherein the review is considered intermittent review.

139. The method of embodiment 135, wherein improvement in or reduction in severity of double vision continues for at least 20 weeks after cessation of antibody administration.

140. The method of embodiment 135, wherein improvement in or reduction in severity of double vision continues for at least 50 weeks after cessation of antibody administration.

141. The method of any one of embodiments 115-140, wherein the antibody is administered as a first dose at a dose of about 1mg/kg to about 5mg/kg antibody.

142. The method of any one of embodiments 115-140, wherein the antibody is administered as a first dose at a dose of about 5mg/kg to about 10mg/kg of antibody.

143. The method of any one of embodiments 115-140, wherein the antibody is administered as a subsequent dose at a dose of about 5mg/kg to about 20mg/kg of antibody.

144. The method of any one of embodiments 115 to 140, wherein the antibody is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses.

145. The method of embodiment 144, wherein the subsequent dose is administered every three weeks for at least 21 weeks.

146. The method of any one of embodiments 115 to 140, wherein the antibody is administered in a pharmaceutical composition comprising a pharmaceutically acceptable diluent, excipient, or carrier.

147. The method of embodiment 146, wherein the pharmaceutical composition further comprises one or more pharmaceutically active compounds for treating TAO.

148. The method of embodiment 146 or 147, wherein the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD 20 antibody; tobrazumab or other anti-IL-6 antibodies; or selenium, infliximab or other anti-tnfα antibodies or Thyroid Stimulating Hormone Receptor (TSHR) inhibitors.

149. A method of increasing IGF-1R internalization on a cell, the method comprising contacting the cell with the antibody or pharmaceutical composition comprising the same of any one of embodiments 107-111.

150. The method of embodiment 149, wherein contacting comprises administering an antibody or a pharmaceutical composition comprising the same to the subject.

151. The method of embodiment 150, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

152. A method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with the antibody of any one of embodiments 107-111 or a pharmaceutical composition comprising the same.

153. The method of embodiment 152, wherein contacting comprises administering to the subject the antibody of any one of embodiments 1-44 or a pharmaceutical composition comprising the same.

154. The method of embodiment 153, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

155. The method of embodiment 153 or 154, wherein the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm.

156. The method of embodiment 155, wherein the IC50 is measured in an in vitro assay, such as the assays provided herein.

157. The method of any one of embodiments 152 to 157, wherein the cell is an a549 cell or an HOCF cell.

158. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the antibody of any one of embodiments 107-111, or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml, or 105 μg/ml for at least 1, 2, or 3 weeks after administration.

159. The method of embodiment 158, wherein the antibody or pharmaceutical composition is administered intravenously.

160. The method of embodiment 158 or 159, wherein the antibody or pharmaceutical composition is administered as a first dose or subsequent dose at a dose of about 1mg/kg to about 5mg/kg (mg antibody/kg subject), about 5mg/kg to about 10mg/kg antibody, or about 5mg/kg to about 20 mg/kg.

161. The method of any one of embodiments 158 to 160, wherein the antibody is administered in the following amounts: about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10mg/kg antibody as a first dose; and about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20mg/kg antibody as a subsequent dose.

162. The method of any one of embodiments 158-161, wherein the antibody or pharmaceutical composition is administered at least or about once a week, once every two weeks, once every 3 weeks, or once every 4 weeks.

163. A method of inhibiting IGF-1 induced receptor autophosphorylation in a subject in need thereof by at least 95%, 96%, 97%, 98%, or 99% or 100%, comprising administering to the subject the antibody of any one of embodiments 107-111 or a pharmaceutical composition comprising the same.

164. A pharmaceutical composition comprising an antibody for treating thyroid eye disease in a subject, wherein the antibody comprises a light chain variable region having the amino acid sequence of SEQ ID No. 13 and a heavy chain variable region having the amino acid sequence of SEQ ID No. 14.

165. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region having M428L and N434S substitutions.

166. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region having M428L, N434S, M Y, S T and T256E substitutions.

167. The pharmaceutical composition of embodiment 164, wherein the antibody comprises an Fc region having M252Y, S254T and T256E substitutions.

168. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID No. 93 and a heavy chain amino acid sequence of SEQ ID No. 92.

169. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID No. 93 and a heavy chain amino acid sequence of SEQ ID No. 94.

170. The pharmaceutical composition of embodiment 164, wherein the antibody comprises a light chain having the amino acid sequence of SEQ ID No. 93 and a heavy chain amino acid sequence of SEQ ID No. 95.

171. A method of treating thyroid eye disease in a subject, the method comprising administering the pharmaceutical composition comprising an antibody of any one of embodiments 164-170.

172. The method of embodiment 171, wherein the pharmaceutical composition is administered intravenously.

173. The method of embodiment 171, wherein the pharmaceutical composition is administered subcutaneously.

174. A method of treating or lessening the severity of thyroid-associated eye disease (TAO) or a symptom thereof in a subject, comprising administering to a subject the pharmaceutical composition of any one of embodiments 164-170.

175. A method of reducing the ocular bulge in a subject's eye with a thyroid-associated ocular disease (TAO), the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

176. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 2-4.

177. A method of reducing the Clinical Activity Score (CAS) of thyroid-related eye disease (TAO) in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

178. A method of reducing a) the bulge eye by at least 2mm and b) the Clinical Activity Score (CAS) in a subject having a thyroid-associated eye disease (TAO), the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

179. The method of any one of embodiments 174 to 178, wherein the protrusion is reduced by at least 2mm.

180. The method of any one of embodiments 174 to 178, wherein the protrusion is reduced by at least 3mm.

181. The method of any one of embodiments 174 to 178, wherein the protrusion is reduced by at least 4mm.

182. The method of any one of embodiments 174 to 178, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

183. The method of any one of embodiments 174 to 178, wherein the subject's Clinical Activity Score (CAS) is reduced to one (1).

184. The method of any one of embodiments 174 to 178, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

185. A method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject comprising administering to a subject the pharmaceutical composition of any one of embodiments 164-170, wherein treatment with said antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

186. A method of improving the quality of life of a subject suffering from thyroid-associated ocular disease (TAO, also known as graves' eye disease), the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

187. The method of embodiment 186, wherein quality of life is measured by a graves' eye disease quality of life (GO-QoL) assessment or a visual function or appearance subscale thereof.

188. The method of embodiment 186, wherein the treatment results in a GO-QoL improvement of greater than or equal to 8 points.

189. The method of embodiment 186, wherein the treatment results in an improvement in the GO-QoL function sub-scale.

190. The method of embodiment 186, wherein the treatment results in an improvement in the appearance subscale of GO-QoL.

191. A method of treating or lessening the severity of a double vision in a subject having a thyroid-associated eye disease (TAO), the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

192. The method of embodiment 191, wherein the review is a constancy review.

193. The method of embodiment 191, wherein the review is a non-constant review.

194. The method of embodiment 191, wherein the review is considered intermittent review.

195. The method of embodiment 191, wherein improvement of double vision or reduction in severity thereof continues for at least 20 weeks after cessation of antibody administration.

196. The method of embodiment 191, wherein improvement of double vision or reduction in severity thereof continues for at least 50 weeks after cessation of antibody administration.

197. The method of any one of embodiments 171 to 196, wherein the pharmaceutical composition is administered as the first dose at a dose of about 1mg/kg to about 5mg/kg, about 5mg/kg to about 10mg/kg, about 10mg/kg to about 20mg/kg, about 20mg/kg to about 30mg/kg, about 5mg/kg, about 10mg/kg, about 15mg/kg, about 20mg/kg, about 25mg/kg, or about 30mg/kg of antibody.

198. The method of any one of embodiments 171 to 196, wherein the pharmaceutical composition is administered at a dose of about 10mg/kg to about 20mg/kg of antibody as the first dose.

199. The method of any one of embodiments 171 to 196, wherein the pharmaceutical composition is administered as a subsequent dose at a dose of about 1mg/kg to about 10mg/kg, about 2mg/kg to about 5mg/kg, or about 5mg/kg to about 20mg/kg of antibody.

200. The method of any one of embodiments 171 to 196, wherein the pharmaceutical composition is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses.

201. The method of embodiment 200, wherein the subsequent dose is administered every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks for at least 21 to 52 weeks or more.

202. A method of increasing IGF-1R internalization on a cell, the method comprising contacting the cell with the pharmaceutical composition of any one of embodiments 164-170.

203. The method of embodiment 202, wherein contacting comprises administering to a subject a pharmaceutical composition of any one of embodiments 164 to 170.

204. The method of embodiment 203, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

205. A method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with the pharmaceutical composition of any one of embodiments 164-170.

206. The method of embodiment 205, wherein contacting comprises administering to the subject the pharmaceutical composition of any one of embodiments 164-170.

207. The method of embodiment 206, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

208. The method of any one of embodiments 205 to 207, wherein the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm.

209. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164-170, wherein the serum concentration of antibody in the subject is at least or about 10 μg/ml or 20 μg/ml or 50 μg/ml, 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml or 105 μg/ml for at least 1, 2 or 3 weeks after administration.

210. The method of embodiment 209, wherein the pharmaceutical composition is administered intravenously or subcutaneously.

211. An isolated antibody comprising a light chain having the amino acid sequence of SEQ ID No. 3 and a heavy chain comprising the amino acid sequence of SEQ ID No. 83.

212. An isolated antibody comprising a variable light chain comprising the sequence of SEQ ID No. 98 and a variable heavy chain comprising the sequence of SEQ ID No. 99 and an Fc region comprising M252Y, S T and T256E mutations.

213. An isolated antibody comprising a variable light chain comprising the sequence of SEQ ID No. 98 and a variable heavy chain comprising the sequence of SEQ ID No. 99 and an Fc region comprising the M428L and N434S mutations.

214. A pharmaceutical composition comprising the antibody of any one of embodiments 211-213.

215. A pharmaceutical composition suitable for intravenous administration comprising the antibody of any one of embodiments 211-213.

216. A pharmaceutical composition suitable for subcutaneous administration comprising the antibody of any one of embodiments 211-213.

217. A method of treating thyroid eye disease in a subject, the method comprising administering a pharmaceutical composition comprising an antibody of any one of embodiments 211-213.

218. The method of embodiment 217, wherein the pharmaceutical composition is administered intravenously.

219. The method of embodiment 217, wherein the pharmaceutical composition is administered subcutaneously.

220. A method of treating or lessening the severity of a thyroid-associated eye disease (TAO) or a symptom thereof, comprising administering to a subject the antibody of any one of embodiments 211-213 or a pharmaceutical composition comprising the same.

221. A method of reducing the ocular bulge in a subject with thyroid-associated ocular disease (TAO), comprising administering to the subject the antibody of any one of embodiments 211-213 or a pharmaceutical composition comprising the same.

222. A method of treating thyroid eye disease in a subject comprising administering to the subject the antibody of any one of embodiments 211-213 or a pharmaceutical composition comprising the same.

223. A method of reducing the Clinical Activity Score (CAS) of a thyroid-associated eye disease (TAO) in a subject comprising administering to the subject the antibody of any one of embodiments 211-213, or a pharmaceutical composition comprising the same.

224. A method of reducing a) the bulge eye by at least 2mm and b) the Clinical Activity Score (CAS) in a subject having a thyroid-associated eye disease (TAO) comprising administering to the subject the antibody of any one of embodiments 211-213, or a pharmaceutical composition comprising the same.

225. The method of any one of embodiments 220 to 224, wherein the protrusion is reduced by at least 2mm.

226. The method of any one of embodiments 220 to 224, wherein the protrusion is reduced by at least 3mm.

227. The method of any one of embodiments 220 to 224, wherein the protrusion is reduced by at least 4mm.

228. The method of any one of embodiments 220 to 224, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

229. The method of any one of embodiments 220 to 224, wherein the subject's Clinical Activity Score (CAS) is reduced to one (1).

230. The method of any one of embodiments 220 to 224, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

231. A method of treating or lessening the severity of thyroid-associated ocular disease (TAO) in a subject comprising administering to a subject the antibody of any one of embodiments 1-3, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

232. A method of improving the quality of life of a subject suffering from thyroid-associated ocular disease (TAO, also known as graves 'eye disease/graves' orbit disease) comprising administering to the subject the antibody of any one of embodiments 211 to 213, or a pharmaceutical composition comprising the same.

233. The method of embodiment 232, wherein quality of life is measured by a graves' eye disease quality of life (GO-QoL) assessment or a visual function or appearance subscale thereof.

234. The method of embodiment 232, wherein the treatment results in a GO-QoL improvement of greater than or equal to 8 points.

235. The method of embodiment 232, wherein the treatment results in an improvement in the GO-QoL functional sub-scale.

236. The method of embodiment 232, wherein the treatment results in an improvement in the appearance subscale of GO-QoL.

237. A method of treating or lessening the severity of a double vision in a subject having a thyroid-associated eye disease (TAO), comprising administering to the subject the antibody of any one of embodiments 211-213, or a pharmaceutical composition comprising the same.

238. The method of embodiment 237, wherein the review is a constancy review.

239. The method of embodiment 237, wherein the review is a non-constant review.

240. The method of embodiment 237, wherein the review is considered intermittent review.

241. The method of embodiment 237, wherein improvement of double vision or reduction in severity thereof continues for at least 20 weeks after cessation of antibody administration.

242. The method of embodiment 237, wherein improvement of double vision or reduction in severity thereof continues for at least 50 weeks after cessation of antibody administration.

243. The method of any one of embodiments 217-242, wherein the antibody is administered as a first dose at a dose of about 1mg/kg to about 5mg/kg, about 5mg/kg to about 10mg/kg, about 10mg/kg to about 20mg/kg, about 20mg/kg to about 30mg/kg, about 5mg/kg, about 10mg/kg, about 15mg/kg, about 20mg/kg, about 25mg/kg, or about 30mg/kg of antibody.

244. The method of any one of embodiments 217-242, wherein the antibody is administered at a dose of about 10mg/kg to about 20mg/kg of antibody as a first dose.

245. The method of any one of embodiments 217-242, wherein the antibody is administered as a subsequent dose at a dose of about 1mg/kg to about 10mg/kg, about 2mg/kg to about 5mg/kg, or about 5mg/kg to about 20mg/kg of antibody.

246. The method of any one of embodiments 217 to 242, wherein the antibody is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses.

247. The method of embodiment 246, wherein the subsequent dose is administered every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks for at least 21 to 52 weeks or more.

248. A method of increasing IGF-1R internalization on a cell, the method comprising contacting the cell with the antibody or pharmaceutical composition comprising the same of any one of embodiments 211-213.

249. The method of embodiment 248, wherein contacting comprises administering an antibody or pharmaceutical composition comprising the same to a subject.

250. The method of embodiment 249, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

251. A method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with the antibody or pharmaceutical composition comprising the same of any one of embodiments 211-213.

252. The method of embodiment 251, wherein contacting comprises administering to the subject an antibody or pharmaceutical composition comprising the same of any one of embodiments 211-213.

253. The method of embodiment 252, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

254. The method of any one of embodiments 251 to 253, wherein the antibody has an IC50 less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm.

255. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the antibody of any one of embodiments 211-213 or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 10 μg/ml or 20 μg/ml or 50 μg/ml, 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml or 105 μg/ml for at least 1, 2 or 3 weeks after administration.

256. The method of embodiment 255, wherein the antibody or the pharmaceutical composition is administered intravenously or subcutaneously.

The subject matter will now be described with reference to the following examples. These embodiments are provided for illustrative purposes only and the claims should in no way be construed as limited to these embodiments, but should be construed to cover any and all variations that become apparent from the teachings provided herein. Those skilled in the art will readily recognize a variety of non-critical parameters that may be changed or modified to produce substantially similar results.

Examples

Example 1: IGF-1R antibodies block IGF-1 stimulation.

Blockade of IGF-1 stimulation is measured by hyaluronic acid secretion in the presence of IGF-1R antibodies VRDN-2700, VRD N-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein. Immunoglobulins were purified from serum of patients with graves' eye disease (GO) and tested for their ability to directly activate TSHR/IGF-1R crosstalk (cross talk) in primary cultures of TSH R and/or IGF-1R and GO fibroblasts. Cells are treated with M22 or GO-Ig with or without IGF-1R inhibiting antibodies (such as those provided herein), including, but not limited to, VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein. Hyaluronic Acid (HA) secretion was measured as the primary biological response of GO fibroblast stimulation. IGF-1R autophosphorylation is used as a measure of direct IGF-1R activation. TSHR activation was determined by cyclic AMP (cAMP) production. IGF-1R antibodies as disclosed herein were found to effectively block HA secretion, and thus were found to block IGF stimulation.

Example 2: treatment of patients with thyroid eye disease and clinical assessment of thyroid eye disease by IGF-1R antibodies.

Infusion of IGF-1R inhibiting antibodies (such as those provided herein) including, but not limited to, VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all of which are disclosed herein, is provided to a subject. The number of infusions was personalized for each subject and based on the clinical judgment of the investigator. The day 1 interrogation occurred within 14 days after the final interrogation of the previous trial. The interrogation window at week 1 and week 4 is + -1 day, and at week 3, week 6, week 9, week 12, week 15, week 18, week 21 and week 24 are + -3 days. The follow-up period was only for subjects who were male eye non-responders in the previous trial; subjects who relapsed in the previous trial did not participate in the follow-up period. The window of inquiry during the follow-up period was + -7 days.

The treatment period was 24 weeks (6 months) during which 8 infusions of tetuzumab were administered.

Subjects as male eye non-responders were scheduled to participate in a 6 month follow-up period in the present extended study; subjects who relapsed in the lead-in study and who were retreated in the present extension study will not participate in the follow-up period.

Efficacy assessment was performed on both eyes at each assessment time point. The "study eye" (i.e., the more severely affected eye) will remain the same as the eye identified at the baseline (day 1) visit of the prior study. Efficacy was assessed for both eyes, but the study eyes were used to assess the primary outcome measure.

Efficacy was assessed by bulge eye (measured as a bulge eye assessment of clinical severity measures using a Hertel instrument to ensure consistency of the measurements), CAS (7-item scale), review (measured as part of clinical severity measure), and clinical severity measures (including movement limitation assessment).

Quality of life was assessed using a GO-QoL questionnaire.

Safety was assessed by AE and concomitant drug use monitoring, immunogenicity testing, physical and ophthalmic examinations, vital signs, clinical safety laboratory evaluations (whole blood count, chemistry (including thyrocheck and HbA 1C) and urinalysis), pregnancy testing (if applicable) and Electrocardiogram (ECG). Study was also monitored by the data security monitoring committee (DSMB).

The bulge eye assessment was performed using a Hertel bulge eye meter to ensure consistency of measurement, and the same Hertel instrument and the same observer (unless unavoidable) were used at each assessment over the complete duration of the study. In addition, the same canthus spacing (ICD) was used in each case.

The convex eye of each eye was measured on days 1 and 6, 12, 18 and 24 (or Premature Withdrawal (PW)) during the treatment period and on months 7, 9 and 12 (or PW) during the follow-up period. Measurements were recorded on the clinical severity measure eCRF of the eyes.

As provided herein, antibodies were found to be effective in treating thyroid eye disease and improving quality of life.

Example 3: antibodies with increased pK

The cynomolgus macaque is administered an amount of 10mg/kg by intravenous or subcutaneous route of antibodies comprising CDRs of VRDN-2700 with YTE mutations in the Fc domain, and samples are collected at time points of 0.5 hours, 2 hours, 8 hours and day 1, day 3, day 7, day 10, day 14, day 21 and day 28 for PK analysis by ELISA. Tituzumab was also administered at 10mg/kg IV as a comparator. The results shown in fig. 1 demonstrate that the antibodies have significantly higher PK as compared to tetuzumab.

This result suggests that antibodies comprising the CDRs of VRDN-2700 are likely to be administered at lower doses than tetuzumab even when administered subcutaneously. These results have not been predicted.

Example 4:

VRDN-1100 is an antagonist antibody against the insulin-like growth factor-1 receptor (IGF-1R) being developed for the treatment of Thyroid Eye Disease (TED). TED is driven by Thyroid Stimulating Hormone Receptor (TSHR) agonistic autoantibodies and cross-talk between TSHR and IGF-1R. TED is characterized by recruitment of fibroblasts expressing IGF-1R and TSHR into the orbital tissues, where they mediate deposition of hyaluronic acid and expansion of orbital muscles and fat 1. IGF-1R antagonism was found to reverse this orbital tissue expansion and firmly alleviate symptom 2 in TED patients.

VRDN-1100 is a humanized monoclonal antibody targeting IGF-1R. The IGF-1R binding and antagonist characteristics of VRDN-1100 were analyzed.

Method

Surface Plasmon Resonance (SPR): the antibody was captured by immobilization of the anti-Fc and recombinant IGF-1R extracellular domain (ECD) flowed as the analyte. Association and dissociation rate constants (ka and KD, respectively) and equilibrium dissociation constants KD were derived by global fitting of the data to a single-site model.

Epitope grouping (Epitope binding): VRDN-1100 was immobilized on the chip surface by amine coupling and used to capture IGF-1R-ECD, after which tetuzumab was flowed on the chip.

Cell binding: a549 human lung adenocarcinoma cells or primary human choroidal fibroblasts (HOCF) were incubated with different concentrations of VRDN-1100 or tetuzumab. A single dose of 50nm IgG1 isotype control was used as negative control. Unbound antibody was removed by washing and cells were incubated with Alexa Fluor 488-goat anti-human antibody and cell impermeable dye to gate for viable cells. The Median Fluorescence Intensity (MFI) of living cells was measured by flow cytometry and the data was analyzed using FlowJo software. Fitting a dose curve using a nonlinear regression model; log (agonist) versus response-variable slope (four parameters).

Internalizing: cells were incubated with different concentrations of antibody of interest at 4℃and 37℃for 60 minutes. Cells were then washed 3 times and incubated with FITC-labeled goat anti-human Fc secondary antibody for 30 minutes at 4 ℃. The MFI of living cells was measured by flow cytometry and the data was analyzed using FlowJo software.

Cell surface marker expression: HOCF cells were incubated with 10ug/mL of directly labeled antibody or IgG isotype control. Median Fluorescence Intensity (MFI) was measured by flow cytometry and data was analyzed using FlowJo software.

Antagonism: serum starved a549 or HOCF cells were preincubated with varying concentrations of test antibody for one hour at 37 ℃ followed by 7 minutes at 37 ℃ by addition of 100ng/mL (a 549) or 200ng/mL (HOCF) IGF-1. Phosphorylated IGF-1R (pIGF 1R) of biological replicates was measured according to the manufacturer's protocol using an R & D Systems pIGF-1R ELISA, and pIGF-1R concentrations were normalized to the lowest test antibody concentration. Fitting a dose curve using a nonlinear regression model; log (inhibitor) vs response-variable slope (four parameters).

Results

VRDN-1100 binds IGF-1R with sub-nanomolar affinity. FIG. 2A illustrates that increasing concentrations of IGF-1R-ECD bound to anti-FC captured VRDN-1100 or tetuzumab revealed a gradual increase in SPR signal, enabling global fitting to a binding model. VRDN-1100 showed more sustained binding interactions after IGF-1R clearance. FIG. 2B illustrates that IGF-1R-ECD binds firmly to immobilized VRDN-1100. Tetuzumab does not exhibit binding to the IGF-1r: VRDN-1100 complex, indicating that tetuzumab and VRDN-1100 have overlapping epitopes. The data is also illustrated in a table as shown in fig. 2.

VRDN-1100 binds IGF-1R on A549 cells with high affinity. As shown in fig. 3, binding of VRDN-1100 to a549 cells was assessed by flow cytometry and found to have a similar binding profile as tetuzumab at three different concentrations. As also shown in fig. 3, the binding dose response curve shows VRDN-1100EC50 = 0.1nM. As shown in figure 3, VRDN-1100, VRDN-2700 with M252Y, S T and T256E mutations in the Fc domain, and tetuzumab display similar binding at temperatures that block IGF-1R receptor internalization. Panel D shows that VRDN-1100, VRDN-2700 with M252Y, S254T and T256E mutations in the Fc domain, and tetuzumab caused similar levels of internalization (about 50%), as measured by a decrease in membrane IGF-1R receptor levels at 37℃versus 4 ℃. In the bar graph of fig. 3, the leftmost bar graph is a isotype control, the left two bar graph sets are tetuzumab, the right two bar graph sets are VRDN-1100 and the rightmost bar graph set is VRDN-2700.

HOCF as an in vitro model of TED pathology.

Cd34+, thy-1+ orbital fibroblasts are involved in extracellular matrix deposition and pathogenic fibrosis in TED 5. As shown in FIG. 4, HOFC expression of (A) IGF-1R and (B) TSHR, and (C) CD34 and Thy-1 was shown, demonstrating its ability to function as an in vitro model system for IGF-1R function in TEDs.

VRDN-1100 binds IGF-1R on HOCF cells with high affinity.

Figure 5 illustrates binding of VRDN-1100 to HOCF cells, which were assessed by flow cytometry and found to have a largely similar binding to tetuzumab at three different concentrations. The graph in the lower right hand corner of fig. 5 shows a binding dose response curve, which demonstrates that EC50 = 0.4nM for VRDN-1100.

VRDN-1100 is a sub-nanomolar IGF-1R antagonist. VRDN-1100 potently inhibited IGF-1 stimulated receptor phosphorylation on a549 cells (ic50=0.09 nM) and HOCF cells (ic50=0.09 nM), which is shown in panels a and B of fig. 6.

These results demonstrate that VRDN-1100 on IGF-1R overlaps with the tetuzumab epitope, VRDN-1100 binds to IGF-1R on cells with a sub-nanomolar EC50, VRDN-1100 promotes IGF-1R internalization, and VRDN-1100 inhibits IGF-1R phosphorylation with a sub-nanomolar IC 50. Thus, VRDN-1100 binds, antagonizes, and internalizes IGF-1R at sub-nanomolar concentrations, suggesting that VRDN-1100 should be able to be used to potentially potently inhibit TED pathophysiology.

Example 4 VRDN-2700 with M252Y, S254T and T256E mutations in the Fc domain is a novel anti-IGF-1R antibody as described herein that incorporates half-life extending modifications in the Fc region of the antibody and is useful for treating Thyroid Eye Disease (TED). Relative to the commercial IGF-1R antibody tetuzumab, pharmacokinetic (PK) parameters of VRDN-2700 with such Fc mutations were measured in cynomolgus macaques, and PK models were constructed to plan potential human dosing regimens.

TED is an autoimmune condition most commonly associated with graves' disease and hyperthyroidism, but is also found in patients with normal or degraded thyroid function. The ocular disease in TED is driven by Thyroid Stimulating Hormone Receptor (TSHR) agonistic autoantibodies and crosstalk between TSHR and IGF-1R. The pathological reconstruction of the orbit and periorbital tissues produces different manifestations, which may include dry eye, increased tear, localized irritation, eyelid retraction and eventual bulge, compound vision and optic nerve compression with consequent vision loss.

The underlying pathology of TED is activation of the inflammatory cascade within the orbit, mainly due to recruitment of fibroblasts and immune cells. Over-expression of IGF-1R has been demonstrated in the orbit of TED patients and IGF-1R inhibiting antibodies are presumed to disrupt IGF-1R and TSHR crosstalk and attenuate the inflammatory cascade. Indeed, IGF-1R antagonism has been demonstrated to firmly alleviate many of the inflammatory symptoms affecting TED patients.

VRDN-2700 is a monoclonal antibody that inhibits IGF-1 mediated signaling at sub nanomolar titers through IGF-1R and incorporates clinically proven extended half-life Fc modifications (M252Y, S254T and T256E). This antibody was found to have a more favorable PK profile than conventional IgG therapeutic antibodies, possibly a less burdened therapeutic modality for the patient.

The cynomolgus macaque was administered with a VRDN-2700 having an Fc mutation by Intravenous (IV) infusion at 2, 10 and 50mg/kg for 30min and by Subcutaneous (SC) injection at 2 and 10 mg/kg. 10mg/kg of tetomilast was likewise administered by 30min IV infusion. VRDN-2700 and tetuzumab levels in serum were measured using a human IgG-specific ELISA assay. Data were analyzed using the WinNonlin non-compartmental model. Semi-mechanical models incorporating target-mediated drug treatment were constructed using available human and cynomolgus macaque data. The data are described below.

The table and diagram of fig. 7 illustrates a more advantageous PK profile.

The table shows PK parameters +/-SD. Evidence of target-mediated drug Treatment (TMDD) was observed at 2mg/kg, but not at the 10 and 50mg/kg doses, consistent with reporting TMDD saturation of tetuzumab and other IGF-1R antibodies at higher doses.

VRDN-2700 half-life extension modification extension exposure

At equivalent doses, SC-administered VRDN-2700 with YTE mutations had greater exposure than intravenous infusion of tetuzumab and reached about 2 times the half-life of tetuzumab in NHP, with the bioavailability (F) of SC-administered VRDN-2700 estimated to be 62% using the preliminary discovery phase formulation. Parameter estimation +/-SD is shown in FIG. 8.

Model simulations predict that administration of VRDN-2700 at 10mg/kg every 3 weeks or 20mg/kg every 6 weeks will result in a Cmin of >100ug/mL, similar to approved tetuzumab regimens (10 mg/kg first dose followed by seven 20mg/kg doses q3 w). The 10mg/kg q3w regimen will have a lower Cmax value. Longer dosing intervals will increase patient convenience and reduce treatment costs, while lower doses and Cmax values can potentially reduce toxicity. Furthermore, the model predicts that a steady state Cmin of about 130ug/mL can be achieved by subcutaneous 300mg fixed dose of VRDN-2700 given weekly, enabling home administration. With lower Cmin values being effective, 300mg fixed dose VRDN-2700 was predicted to reach steady state Cmin levels of about 50ug/mL every other week. In summary, the extended half-life of VRDN-2700 is predicted to provide patients with a wider selection of more appropriate dosing intervals and routes of administration.

Example 5: VRDN2700 properties during evaluation of antibodies, expression of VRDN-2700 was compared to other antibodies having mutations in the Fc domain (L/S mutations as described herein). Unexpectedly, the yield of antibodies with YTE mutations in the Fc domain (VRDN 2700) was approximately 80% higher than the yield of similar antibodies with L/S mutations. This is surprising and unexpected because other antibodies that have been tested for targeting IGF-1R with YTE or LS mutations have similar expression regardless of Fc mutation. The YTE version has fewer low molecular weight species than the LS version. Thus, it was shown that YTE antibodies have fewer impurities and are more homogeneous compositions, which provide advantages over antibodies with LS mutations. This is also unpredictable, as the other antibody evaluated exhibits an adverse effect on such species. Furthermore, during purification, LS mutants were found to form more aggregates than VRDN-2700 when purified on cation exchange columns. Aggregation of LS mutants would cause high volume manufacturing problems that were not observed for VRDN-2700. Thus, such differences in Fc mutants against this antibody may not have been predicted or expected and produce a number of and unexpected advantages for the antibody referred to herein as VRDN-2700.

The extended half-life of VRDN-2700 (YTE) demonstrates that it can be used for convenient SC injection, or as IV infusion, as compared to conventional therapeutic IgG antibodies, requires less and/or less frequent treatment, and has superior properties compared to other Fc mutant versions of the same antibody (the same variable region).

Example 6: VRDN-1100 with YTE or YTE/C22S mutations binds to IGF-1R and inhibits IGF-1R autophosphorylation. Binding of VRDN-1100 with Fc YTE mutation in the heavy chain (SEQ ID NO: 94) or with C22S mutation and Fc YTE mutation in the heavy chain (SEQ ID NO: 95) to IGF-1R was evaluated in a cell-based binding assay (A549 cells). The light chain has the sequence of SEQ ID NO. 93. It was found that the YTE Fc mutant of VRDN1100 bound to a549 cells with an EC50 of 0.30nm and that the C22S and Fc YTE mutants had an EC50 of 0.36 nm. Antibodies were also evaluated for their ability to inhibit IGF-1R autophosphorylation. Only the YTE mutant had an IC50 of 0.40nm and the C22S plus YTE mutation had an IC50 of 0.37 nm. Thus, antibodies were found to bind both to IGF-1R and to inhibit autophosphorylation.

Example 7: VRDN-1100 with C22S mutation binds to IGF-1R. Mutants of VRDN-1100 having a C22S mutation in the heavy chain (SEQ ID NO: 96) and VL comprising the sequence of SEQ ID NO:97 were evaluated for binding to IGF-1R in a surface plasmon resonance assay. Using this assay, the antibodies were found to be 1.04X 10 at pH 7.4 ⁵ K of (2) _a (1/Ms)、2.18×10 ^-5 K of (2) _d (1/s) and 2.10X10 ^-10 K of (2) _D (M) binds to IGF-1R.

Each of these examples and embodiments provided herein demonstrate that the antibodies provided herein can be used to treat TED and its associated symptoms.

All references cited herein are incorporated by reference as if each individual publication, database entry (e.g., genbank sequence or GeneID entry), patent application or patent were specifically and individually indicated to be incorporated by reference. Applicant follows 37c.f.r. ≡1.57 (b) (1), the expressions incorporated by reference herein are with respect to each individual publication, database entry (e.g., genbank sequence or GeneID entry), patent application, or patent, each of which is specifically identified in accordance with 37c.f.r. ≡1.57 (b) (2), even though such references do not approximate the specific expressions incorporated by reference. The inclusion of a specific expression, if any, incorporated by reference herein does not in any way weaken the general expression incorporated by reference herein. Citation of references herein is not intended as an admission that such references are pertinent background, nor does it constitute any admission as to the contents or date of such publications or documents.

The scope of embodiments of the invention is not limited by the specific embodiments described herein. Indeed, various modifications of the described modes for carrying out the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the embodiments and any appended claims.

The present description is deemed sufficient to enable one skilled in the art to practice the embodiments. Various modifications other than those shown and described herein will be apparent to those skilled in the art from the foregoing description and are within the scope of the present disclosure and any appended claims.

Sequence listing

<110> Weili dean treatment Co (Viridian Therapeutics, inc.)

<120> compositions and methods for treating thyroid eye disorders

<130> 257635.000112

<150> 63/091839

<151> 2020-10-14

<150> 63/201978

<151> 2021-05-21

<150> 63/260130

<151> 2021-08-10

<150> 63/261742

<151> 2021-09-28

<160> 99

<170> PatentIn version 3.5

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

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

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

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

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

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Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Lys Trp Pro Pro

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

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

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Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

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

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

165 170 175

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

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Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

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Ser Phe Asn Arg Gly Glu Cys

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

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

50 55 60

Arg 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 Phe Cys

85 90 95

Ala Arg Glu Leu Gly Arg Arg Tyr Phe Asp Leu Trp Gly Arg Gly Thr

100 105 110

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

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

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

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Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

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

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Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

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His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

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

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Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

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

275 280 285

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

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

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

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

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

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Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro

50 55 60

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

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Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

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

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Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Gly Gly

20 25 30

Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

35 40 45

Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu

50 55 60

Lys Asp Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

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

100 105 110

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

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

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

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

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

195 200 205

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

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

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

275 280 285

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

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

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

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

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Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

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

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

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His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

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

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Lys Ser Arg Asp Gly Ser Gly Gln His

85 90 95

Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys

100 105 110

Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser

180 185 190

Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val

195 200 205

Ala Pro Ala Glu Cys Ser

210

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Glu Val Gln Leu Val 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

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Ala Arg Ala Pro Leu Arg Phe Leu Glu Trp Ser Thr Gln Asp His Tyr

100 105 110

Tyr Tyr Tyr Tyr Met Asp Val Trp Gly Lys Gly Thr Thr Val Thr Val

115 120 125

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

130 135 140

Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys

145 150 155 160

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu

165 170 175

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

180 185 190

Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr

195 200 205

Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val

210 215 220

Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro

225 230 235 240

Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe

245 250 255

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

260 265 270

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe

275 280 285

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

290 295 300

Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

305 310 315 320

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

325 330 335

Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala

340 345 350

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg

355 360 365

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

370 375 380

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

385 390 395 400

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

405 410 415

Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln

420 425 430

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

435 440 445

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

450 455 460

<210> 7

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Asp Ile Gln Met Thr Gln Phe 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 Arg Leu His Arg 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 Cys

85 90 95

Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 8

<211> 450

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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 Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

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

35 40 45

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

50 55 60

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys

195 200 205

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

210 215 220

Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala

225 230 235 240

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

245 250 255

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

260 265 270

Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val

275 280 285

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Pro Gly

450

<210> 9

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

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

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly

85 90 95

Thr His Trp Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 10

<211> 449

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser

20 25 30

Asn Trp Trp Ser Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

35 40 45

Ile Gly Glu Ile Tyr His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Val Thr Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

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

100 105 110

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

115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

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

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

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

195 200 205

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys

210 215 220

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

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

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

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

405 410 415

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

Lys

<210> 11

<211> 214

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 11

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

1 5 10 15

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

20 25 30

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

35 40 45

Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg Leu Pro His

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 12

<211> 448

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 12

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

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

85 90 95

Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr

100 105 110

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

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

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

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

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

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

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

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

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

275 280 285

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

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

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

385 390 395 400

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

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 13

<211> 113

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 13

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg

<210> 14

<211> 124

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 14

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

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

115 120

<210> 15

<211> 107

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 15

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Asp Ser Leu Pro His

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 16

<211> 120

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 16

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 Ile Tyr

20 25 30

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

35 40 45

Ser Gly Ile Ser Pro Ser Gly Gly Thr Thr Trp 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 Ser Gly Gly Ser Gly Tyr Ala Phe Asp Ile Trp Gly Gln

100 105 110

Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 17

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 17

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala

1 5 10

<210> 18

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 18

Asp Ala Ser Lys Arg Ala Thr

1 5

<210> 19

<211> 10

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 19

Gln Gln Arg Ser Lys Trp Pro Pro Trp Thr

1 5 10

<210> 20

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 20

Ser Tyr Gly Met His

1 5

<210> 21

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 21

Ile Ile Trp Phe Asp Gly Ser Ser Thr Tyr Tyr Ala Asp Ser Val Arg

1 5 10 15

Gly

<210> 22

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 22

Glu Leu Gly Arg Arg Tyr Phe Asp Leu

1 5

<210> 23

<211> 20

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 23

Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr Leu Gln

1 5 10 15

Trp Tyr Leu Gln

20

<210> 24

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 24

Lys Val Ser Asn Arg Leu Tyr

1 5

<210> 25

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 25

Phe Gln Gly Ser His Val Pro Trp Thr

1 5

<210> 26

<211> 6

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 26

Gly Gly Tyr Leu Trp Asn

1 5

<210> 27

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 27

Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu Lys Asp

1 5 10 15

<210> 28

<211> 8

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 28

Tyr Gly Arg Val Phe Phe Asp Tyr

1 5

<210> 29

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 29

Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Thr

1 5 10

<210> 30

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 30

Gly Glu Asn Lys Arg Pro Ser

1 5

<210> 31

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 31

Lys Ser Arg Asp Gly Ser Gly Gln His Leu Val

1 5 10

<210> 32

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 32

Ser Tyr Ala Ile Ser

1 5

<210> 33

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 33

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

1 5 10 15

Gly

<210> 34

<211> 21

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 34

Ala Pro Leu Arg Phe Leu Glu Trp Ser Thr Gln Asp His Tyr Tyr Tyr

1 5 10 15

Tyr Tyr Met Asp Val

20

<210> 35

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 35

Arg Ala Ser Gln Gly Ile Arg Asn Asp Leu Gly

1 5 10

<210> 36

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 36

Ala Ala Ser Arg Leu His Arg

1 5

<210> 37

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 37

Leu Gln His Asn Ser Tyr Pro Cys Ser

1 5

<210> 38

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 38

Ser Tyr Ala Met Asn

1 5

<210> 39

<211> 17

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 39

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

1 5 10 15

Gly

<210> 40

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 40

Asp Leu Gly Trp Ser Asp Ser Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val

1 5 10 15

<210> 41

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 41

Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp

1 5 10 15

<210> 42

<211> 6

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 42

Leu Gly Ser Asn Arg Ala

1 5

<210> 43

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 43

Met Gln Gly Thr His Trp Pro Leu Thr

1 5

<210> 44

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 44

Ser Ser Ser Asn Trp Trp Ser

1 5

<210> 45

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 45

Glu Ile Tyr His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser

1 5 10 15

<210> 46

<211> 10

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 46

Trp Thr Gly Arg Thr Asp Ala Phe Asp Ile

1 5 10

<210> 47

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 47

Arg Ala Ser Gln Ser Ile Gly Ser Ser Leu His

1 5 10

<210> 48

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 48

Tyr Ala Ser Gln Ser Leu Ser

1 5

<210> 49

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 49

His Gln Ser Ser Arg Leu Pro His Thr

1 5

<210> 50

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 50

Ser Phe Ala Met His

1 5

<210> 51

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 51

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

1 5 10 15

<210> 52

<211> 10

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 52

Leu Gly Asn Phe Tyr Tyr Gly Met Asp Val

1 5 10

<210> 53

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 53

Arg Ser Ser Gln Ser Ile Val His Ser Asn Val Asn Thr Tyr Leu Glu

1 5 10 15

<210> 54

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 54

Lys Val Ser Asn Arg Phe Ser

1 5

<210> 55

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 55

Phe Gln Gly Ser His Val Pro Pro Thr

1 5

<210> 56

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 56

Ser Tyr Trp Met His

1 5

<210> 57

<211> 18

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 57

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

1 5 10 15

Gln Gly

<210> 58

<211> 15

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 58

Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp Val

1 5 10 15

<210> 59

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 59

Gln Ala Ser Arg Asp Ile Arg Asn Tyr Leu Asn

1 5 10

<210> 60

<211> 7

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 60

Asp Ala Ser Ser Leu Gln Thr

1 5

<210> 61

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 61

Gln Gln Phe Asp Ser Leu Pro His Thr

1 5

<210> 62

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 62

Ile Tyr Arg Met Gln

1 5

<210> 63

<211> 16

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 63

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

1 5 10 15

<210> 64

<211> 11

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 64

Trp Ser Gly Gly Ser Gly Tyr Ala Phe Asp Ile

1 5 10

<210> 65

<211> 663

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 65

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Lys Trp Pro Pro

85 90 95

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

100 105 110

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

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys Gln Val Glu Leu Val Glu Ser Gly Gly

210 215 220

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

225 230 235 240

Gly Phe Thr Phe Ser Ser Tyr Gly Met His Trp Val Arg Gln Ala Pro

245 250 255

Gly Lys Gly Leu Glu Trp Val Ala Ile Ile Trp Phe Asp Gly Ser Ser

260 265 270

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

275 280 285

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

290 295 300

Asp Thr Ala Val Tyr Phe Cys Ala Arg Glu Leu Gly Arg Arg Tyr Phe

305 310 315 320

Asp Leu Trp Gly Arg Gly Thr Leu Val Ser Val Ser Ser Ala Ser Thr

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro

435 440 445

Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

450 455 460

Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

465 470 475 480

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

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

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

565 570 575

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

580 585 590

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

595 600 605

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

610 615 620

Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser

625 630 635 640

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

645 650 655

Leu Ser Leu Ser Pro Gly Lys

660

<210> 66

<211> 666

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 66

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro

50 55 60

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

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln

210 215 220

Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr

225 230 235 240

Cys Thr Val Ser Gly Tyr Ser Ile Thr Gly Gly Tyr Leu Trp Asn Trp

245 250 255

Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Tyr Ile Ser

260 265 270

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

275 280 285

Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser

290 295 300

Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Tyr Gly Arg

305 310 315 320

Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

325 330 335

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

340 345 350

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

355 360 365

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

370 375 380

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

385 390 395 400

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

405 410 415

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

420 425 430

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

435 440 445

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

450 455 460

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

465 470 475 480

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

485 490 495

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

500 505 510

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

515 520 525

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

530 535 540

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

545 550 555 560

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

565 570 575

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

580 585 590

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

595 600 605

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

610 615 620

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

625 630 635 640

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

645 650 655

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

660 665

<210> 67

<211> 674

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 67

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Asn Lys Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser

50 55 60

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

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Lys Ser Arg Asp Gly Ser Gly Gln His

85 90 95

Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys

100 105 110

Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser

180 185 190

Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val

195 200 205

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

210 215 220

Val Lys Lys Pro Gly Ser Ser Val Lys Val Ser Cys Lys Ala Ser Gly

225 230 235 240

Gly Thr Phe Ser Ser Tyr Ala Ile Ser Trp Val Arg Gln Ala Pro Gly

245 250 255

Gln Gly Leu Glu Trp Met Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala

260 265 270

Asn Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Ile Thr Ala Asp Lys

275 280 285

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

290 295 300

Thr Ala Val Tyr Tyr Cys Ala Arg Ala Pro Leu Arg Phe Leu Glu Trp

305 310 315 320

Ser Thr Gln Asp His Tyr Tyr Tyr Tyr Tyr Met Asp Val Trp Gly Lys

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

405 410 415

Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys

420 425 430

Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp

435 440 445

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

450 455 460

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile

465 470 475 480

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

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

565 570 575

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

580 585 590

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

595 600 605

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

610 615 620

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

625 630 635 640

Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His

645 650 655

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

660 665 670

Gly Lys

<210> 68

<211> 664

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 68

Asp Ile Gln Met Thr Gln Phe 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 Arg Leu His Arg 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 Cys

85 90 95

Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Leu Glu Ser Gly Gly Gly

210 215 220

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

225 230 235 240

Phe Thr Phe Ser Ser Tyr Ala Met Asn Trp Val Arg Gln Ala Pro Gly

245 250 255

Lys Gly Leu Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Thr Thr

260 265 270

Phe Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn

275 280 285

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

290 295 300

Thr Ala Val Tyr Tyr Cys Ala Lys Asp Leu Gly Trp Ser Asp Ser Tyr

305 310 315 320

Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr

325 330 335

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

340 345 350

Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val

355 360 365

Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala

370 375 380

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

385 390 395 400

Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly

405 410 415

Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys

420 425 430

Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys

435 440 445

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

450 455 460

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

465 470 475 480

Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr

485 490 495

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

500 505 510

Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His

515 520 525

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

530 535 540

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

545 550 555 560

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

565 570 575

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

580 585 590

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

595 600 605

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

610 615 620

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

625 630 635 640

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

645 650 655

Lys Ser Leu Ser Leu Ser Pro Gly

660

<210> 69

<211> 668

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 69

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser

20 25 30

Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro

50 55 60

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

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Gly

85 90 95

Thr His Trp Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln

210 215 220

Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gly Thr Leu Ser Leu Thr

225 230 235 240

Cys Ala Val Ser Gly Gly Ser Ile Ser Ser Ser Asn Trp Trp Ser Trp

245 250 255

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

260 265 270

His Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr

275 280 285

Ile Ser Val Asp Lys Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser

290 295 300

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

305 310 315 320

Arg Thr Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr Val

325 330 335

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

340 345 350

Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys

355 360 365

Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu

370 375 380

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

385 390 395 400

Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr

405 410 415

Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val

420 425 430

Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro

435 440 445

Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe

450 455 460

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

465 470 475 480

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe

485 490 495

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

500 505 510

Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

515 520 525

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

530 535 540

Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala

545 550 555 560

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg

565 570 575

Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

580 585 590

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

595 600 605

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

610 615 620

Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln

625 630 635 640

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

645 650 655

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

660 665

<210> 70

<211> 662

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 70

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

1 5 10 15

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

20 25 30

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

35 40 45

Lys Tyr Ala Ser Gln Ser Leu Ser Gly Ile Pro Asp Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys His Gln Ser Ser Arg Leu Pro His

85 90 95

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

100 105 110

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

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

Phe Asn Arg Gly Glu Cys Glu Val Gln Leu Val Gln Ser Gly Gly Gly

210 215 220

Leu Val Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly

225 230 235 240

Phe Thr Phe Ser Ser Phe Ala Met His Trp Val Arg Gln Ala Pro Gly

245 250 255

Lys Gly Leu Glu Trp Ile Ser Val Ile Asp Thr Arg Gly Ala Thr Tyr

260 265 270

Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala

275 280 285

Lys Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr

290 295 300

Ala Val Tyr Tyr Cys Ala Arg Leu Gly Asn Phe Tyr Tyr Gly Met Asp

305 310 315 320

Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

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

450 455 460

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

465 470 475 480

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

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

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

565 570 575

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

580 585 590

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

595 600 605

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

610 615 620

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

625 630 635 640

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

645 650 655

Ser Leu Ser Pro Gly Lys

660

<210> 71

<211> 673

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 71

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val

210 215 220

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

225 230 235 240

Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val

245 250 255

Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro

260 265 270

Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe Gln Gly Lys Ala Thr

275 280 285

Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser

290 295 300

Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe Ala Arg Gly Arg Pro

305 310 315 320

Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp Val Trp Gly Gln Gly

325 330 335

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

340 345 350

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

355 360 365

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

370 375 380

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

385 390 395 400

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

405 410 415

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

420 425 430

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys

435 440 445

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

450 455 460

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

465 470 475 480

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

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

565 570 575

Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr

580 585 590

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

595 600 605

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

610 615 620

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

625 630 635 640

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

645 650 655

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

660 665 670

Lys

<210> 72

<211> 227

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 72

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

1 5 10 15

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

20 25 30

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

35 40 45

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

50 55 60

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

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Phe Asp Ser Leu Pro His

85 90 95

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

100 105 110

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

115 120 125

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ile Tyr Arg Met Gln Trp Val

130 135 140

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

145 150 155 160

Ser Gly Gly Thr Thr Trp Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr

165 170 175

Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser

180 185 190

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Trp Ser Gly

195 200 205

Gly Ser Gly Tyr Ala Phe Asp Ile Trp Gly Gln Gly Thr Met Val Thr

210 215 220

Val Ser Ser

225

<210> 73

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<220>

<221> MISC_FEATURE

<222> (1)..(5)

<223> n=1-5

<400> 73

Gly Gly Gly Gly Ser

1 5

<210> 74

<211> 5

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<220>

<221> MISC_FEATURE

<222> (1)..(5)

<223> n=1-5

<400> 74

Gly Gly Gly Gly Ala

1 5

<210> 75

<211> 217

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 75

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

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

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

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

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

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

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

100 105 110

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

115 120 125

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

130 135 140

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

145 150 155 160

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

165 170 175

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

180 185 190

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

195 200 205

Lys Ser Leu Ser Leu Ser Pro Gly Lys

210 215

<210> 76

<211> 217

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 76

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

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

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

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

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

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

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

100 105 110

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

115 120 125

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

130 135 140

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

145 150 155 160

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

165 170 175

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

180 185 190

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

195 200 205

Lys Ser Leu Ser Leu Ser Pro Gly Lys

210 215

<210> 77

<211> 215

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 77

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

1 5 10 15

Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val

20 25 30

Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val

35 40 45

Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln

50 55 60

Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln

65 70 75 80

Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly

85 90 95

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

100 105 110

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

115 120 125

Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser

130 135 140

Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr

145 150 155 160

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

165 170 175

Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe

180 185 190

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

195 200 205

Ser Leu Ser Leu Ser Pro Gly

210 215

<210> 78

<211> 232

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 78

Asp Ile Gln Met Thr Gln Phe 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 Arg Leu His Arg 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 Ser

85 90 95

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

100 105 110

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

115 120 125

Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr Ala Met Asn Trp Val

130 135 140

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

145 150 155 160

Ser Gly Gly Thr Thr Phe Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr

165 170 175

Ile Ser Arg Asp Asn Ser Arg Thr Thr Leu Tyr Leu Gln Met Asn Ser

180 185 190

Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Asp Leu Gly

195 200 205

Trp Ser Asp Ser Tyr Tyr Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln

210 215 220

Gly Thr Thr Val Thr Val Ser Ser

225 230

<210> 79

<211> 107

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 79

Asp Ile Gln Met Thr Gln Phe 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 Arg Leu His Arg 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 Ser

85 90 95

Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 80

<211> 125

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<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 Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

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

35 40 45

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

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Arg Thr 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 Leu Gly Trp Ser Asp Ser 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> 81

<211> 9

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 81

Leu Gln His Asn Ser Tyr Pro Ser Ser

1 5

<210> 82

<211> 665

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 82

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro

50 55 60

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

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Gln

210 215 220

Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu Thr Leu Ser Leu Thr

225 230 235 240

Cys Thr Val Ser Gly Tyr Ser Ile Thr Gly Gly Tyr Leu Trp Asn Trp

245 250 255

Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile Gly Tyr Ile Ser

260 265 270

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

275 280 285

Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser

290 295 300

Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Tyr Gly Arg

305 310 315 320

Val Phe Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

325 330 335

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

340 345 350

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

355 360 365

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

370 375 380

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

385 390 395 400

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

405 410 415

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

420 425 430

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

435 440 445

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

450 455 460

Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys

465 470 475 480

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

485 490 495

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

500 505 510

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

515 520 525

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

530 535 540

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

545 550 555 560

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

565 570 575

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

580 585 590

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

595 600 605

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

610 615 620

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

625 630 635 640

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

645 650 655

Gln Lys Ser Leu Ser Leu Ser Pro Gly

660 665

<210> 83

<211> 446

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 83

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Gly Gly

20 25 30

Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

35 40 45

Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu

50 55 60

Lys Asp Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

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

100 105 110

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

115 120 125

Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys

130 135 140

Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser

145 150 155 160

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

165 170 175

Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser

180 185 190

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

195 200 205

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

210 215 220

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

260 265 270

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

275 280 285

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

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

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

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

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

405 410 415

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 84

<211> 329

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 84

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

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

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

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

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

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

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

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu

225 230 235 240

Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

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

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 85

<211> 673

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 85

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Gln Val Gln Leu Val

210 215 220

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

225 230 235 240

Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Trp Met His Trp Val

245 250 255

Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Glu Ile Asn Pro

260 265 270

Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe Gln Gly Lys Ala Thr

275 280 285

Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu Ser Ser

290 295 300

Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe Ala Arg Gly Arg Pro

305 310 315 320

Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp Val Trp Gly Gln Gly

325 330 335

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

340 345 350

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

355 360 365

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

370 375 380

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

385 390 395 400

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

405 410 415

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

420 425 430

Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys

435 440 445

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

450 455 460

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

465 470 475 480

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

485 490 495

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

500 505 510

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

515 520 525

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

530 535 540

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

545 550 555 560

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

565 570 575

Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr

580 585 590

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

595 600 605

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

610 615 620

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

625 630 635 640

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

645 650 655

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

660 665 670

Lys

<210> 86

<211> 113

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 86

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg

<210> 87

<211> 330

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 87

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

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

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

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

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

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

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

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 88

<211> 329

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 88

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

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

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

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

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

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

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

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 89

<211> 329

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 89

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

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

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

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

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

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

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

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly

325

<210> 90

<211> 330

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 90

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

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

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

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

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

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys

100 105 110

Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

115 120 125

Lys Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys

130 135 140

Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp

145 150 155 160

Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu

165 170 175

Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu

180 185 190

His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn

195 200 205

Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly

210 215 220

Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu

225 230 235 240

Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

245 250 255

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

260 265 270

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

275 280 285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295 300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315 320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325 330

<210> 91

<211> 124

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 91

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

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

115 120

<210> 92

<211> 454

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 92

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

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

245 250 255

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Ser Leu Ser Pro Gly Lys

450

<210> 93

<211> 219

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 93

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

115 120 125

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

130 135 140

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

145 150 155 160

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

165 170 175

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

180 185 190

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

195 200 205

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 94

<211> 453

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 94

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Ser Leu Ser Pro Gly

450

<210> 95

<211> 453

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 95

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys

115 120 125

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

130 135 140

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

145 150 155 160

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

165 170 175

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

180 185 190

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

195 200 205

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

210 215 220

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

225 230 235 240

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

245 250 255

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

260 265 270

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

275 280 285

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

290 295 300

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

305 310 315 320

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

325 330 335

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

340 345 350

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

355 360 365

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

370 375 380

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

385 390 395 400

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

405 410 415

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

420 425 430

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

435 440 445

Ser Leu Ser Pro Gly

450

<210> 96

<211> 124

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 96

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

1 5 10 15

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

20 25 30

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

35 40 45

Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Gln Lys Phe

50 55 60

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

65 70 75 80

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Phe

85 90 95

Ala Arg Gly Arg Pro Asp Tyr Tyr Gly Ser Ser Lys Trp Tyr Phe Asp

100 105 110

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

115 120

<210> 97

<211> 112

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 97

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

1 5 10 15

Asp Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Arg Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Ile Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 98

<211> 113

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 98

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

1 5 10 15

Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser

20 25 30

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

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Leu Tyr Gly Val Pro

50 55 60

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

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

Arg

<210> 99

<211> 117

<212> PRT

<213> Artificial sequence (Artificial Sequence)

<220>

<223> synthetic sequence

<400> 99

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

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Gly Gly

20 25 30

Tyr Leu Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp

35 40 45

Ile Gly Tyr Ile Ser Tyr Asp Gly Thr Asn Asn Tyr Lys Pro Ser Leu

50 55 60

Lys Asp Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser

65 70 75 80

Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys

85 90 95

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

100 105 110

Val Thr Val Ser Ser

115

Claims

1. An isolated antibody comprising a light chain having the amino acid sequence of SEQ ID No. 3 and a heavy chain comprising the amino acid sequence of SEQ ID No. 83.

2. An isolated antibody comprising a variable light chain comprising the sequence of SEQ ID No. 98 and a variable heavy chain comprising the sequence of SEQ ID No. 99 and an Fc region comprising M252Y, S T and T256E mutations.

3. An isolated antibody comprising a variable light chain comprising the sequence of SEQ ID No. 98 and a variable heavy chain comprising the sequence of SEQ ID No. 99 and an Fc region comprising the M428L and N434S mutations.

4. A pharmaceutical composition comprising the antibody of any one of claims 1 to 3.

5. A pharmaceutical composition suitable for intravenous administration comprising the antibody of any one of claims 1 to 3.

6. A pharmaceutical composition suitable for subcutaneous administration comprising the antibody of any one of claims 1 to 3.

7. A method of treating thyroid eye disease in a subject, the method comprising administering a pharmaceutical composition comprising the antibody of any one of claims 1 to 3.

8. The method of claim 7, wherein the pharmaceutical composition is administered intravenously.

9. The method of claim 7, wherein the pharmaceutical composition is administered subcutaneously.

10. A method of treating or lessening the severity of thyroid-associated eye disease (TAO) or a symptom thereof, the method comprising administering to a subject the antibody of any one of claims 1-3 or a pharmaceutical composition comprising the same.

11. A method of reducing the ocular bulge in a subject suffering from thyroid-associated ocular disease (TAO), comprising administering to the subject the antibody of any one of claims 1 to 3 or a pharmaceutical composition comprising the same.

12. A method of treating thyroid eye disease in a subject comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same.

13. A method of reducing the Clinical Activity Score (CAS) of a thyroid-associated eye disease (TAO) in a subject, comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same.

14. A method of reducing a) the bulge eye by at least 2mm and b) the Clinical Activity Score (CAS) in a subject suffering from thyroid-associated eye disease (TAO), comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same.

15. The method of any one of claims 10 to 14, wherein the protrusion is reduced by at least 2mm.

16. The method of any one of claims 10 to 14, wherein the protrusion is reduced by at least 3mm.

17. The method of any one of claims 10 to 14, wherein the protrusion is reduced by at least 4mm.

18. The method of any one of claims 10-14, wherein the subject's Clinical Activity Score (CAS) is reduced by at least 2 points.

19. The method of any one of claims 10-14, wherein the subject's Clinical Activity Score (CAS) is reduced to one (1).

20. The method of any one of claims 10-14, wherein the subject's Clinical Activity Score (CAS) is reduced to zero (0).

21. A method of treating or lessening the severity of thyroid-associated eye disease (TAO) in a subject comprising administering to a subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same, wherein treatment with said antibody (i) reduces the relief of one eye by at least 2mm; (ii) Does not accompany the deterioration of the other eye (or the contralateral eye) by 2mm or more; and (iii) reducing CAS of the subject to one (1) or zero (0).

22. A method of improving the quality of life of a subject suffering from thyroid-associated eye disease (TAO, also known as graves 'eye disease/graves' orbital disease), the method comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same.

23. The method of claim 22, wherein the quality of life is measured by a graves' eye disease quality of life (GO-QoL) assessment or a vision function or appearance sub-scale thereof.

24. The method of claim 22, wherein the treatment results in a GO-QoL improvement of greater than or equal to 8 points.

25. The method of claim 22, wherein the treatment results in an improvement in the GO-QoL function sub-scale.

26. The method of claim 22, wherein the treatment results in an improvement in the appearance sub-scale of GO-QoL.

27. A method of treating or lessening the severity of a double vision in a subject suffering from a thyroid-associated eye disease (TAO), the method comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same.

28. The method of claim 27, wherein the complex is regarded as a constant complex view.

29. The method of claim 27, wherein the review is non-constant review.

30. The method of claim 27, wherein the review is considered intermittent review.

31. The method of claim 27, wherein the improvement in or severity of the double vision is reduced for at least 20 weeks after cessation of antibody administration.

32. The method of claim 27, wherein the improvement in or severity of the double vision is reduced for at least 50 weeks after cessation of antibody administration.

33. The method of any one of claims 7 to 32, wherein the antibody is administered as a first dose at a dose of about 1mg/kg to about 5mg/kg, about 5mg/kg to about 10mg/kg, about 10mg/kg to about 20mg/kg, about 20mg/kg to about 30mg/kg, about 5mg/kg, about 10mg/kg, about 15mg/kg, about 20mg/kg, about 25mg/kg, or about 30mg/kg of antibody.

34. The method of any one of claims 7 to 32, wherein the antibody is administered as a first dose at a dose of about 10mg/kg to about 20mg/kg of antibody.

35. The method of any one of claims 7 to 32, wherein the antibody is administered as a subsequent dose at a dose of about 1mg/kg to about 10mg/kg, about 2mg/kg to about 5mg/kg, or about 5mg/kg to about 20mg/kg of antibody.

36. The method of any one of claims 7 to 32, wherein the antibody is administered in the following amounts: about 10mg/kg antibody as the first dose; and about 20mg/kg antibody as subsequent doses.

37. The method of claim 36, wherein the subsequent dose is administered every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks for at least 21 to 52 weeks or more.

38. A method of increasing IGF-1R internalization on a cell, the method comprising contacting the cell with the antibody of any one of claims 1 to 3 or a pharmaceutical composition comprising the same.

39. The method of claim 38, wherein the contacting comprises administering the antibody or a pharmaceutical composition comprising the same to a subject.

40. The method of claim 39, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

41. A method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell, the method comprising contacting the cell with the antibody of any one of claims 1 to 3 or a pharmaceutical composition comprising the same.

42. The method of claim 41, wherein the contacting comprises administering the antibody of any one of claims 1 to 3 or a pharmaceutical composition comprising the same to a subject.

43. The method of claim 42, wherein the subject has or is at risk of Thyroid Eye Disease (TED).

44. The method of any one of claims 41-43, wherein the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09 nm.

45. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject the antibody of any one of claims 1 to 3, or a pharmaceutical composition comprising the same, wherein the serum concentration of the antibody in the subject is at least or about 10 μg/ml or 20 μg/ml or 50 μg/ml, 70 μg/ml, 75 μg/ml, 80 μg/ml, 85 μg/ml, 90 μg/ml, 95 μg/ml, 100 μg/ml or 105 μg/ml for at least 1, 2 or 3 weeks after administration.

46. The method of claim 45, wherein the antibody or the pharmaceutical composition is administered intravenously or subcutaneously.