CN116635024A - Compositions and methods for treating thyroid eye disease - Google Patents

Abstract

The present invention provides anti-IGF-1R antibodies and compositions, and uses thereof.

Description

Compositions and methods for treating thyroid eye disease

CROSS-REFERENCE TO RELATED APPLICATIONS:

This application claims priority to U.S. Provisional Application No. 63/091,839 filed on October 14, 2020, U.S. Provisional Application No. 63/201,978 filed on May 21, 2021, U.S. Provisional Application No. 63/260,130 filed on August 10, 2021, and U.S. Provisional Application No. 63/261,742 filed on September 28, 2021, each of which is incorporated herein by reference in its entirety.

Background technology:

Thyroid-associated eye disease (TAO) (also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmos, dysthyroid ophthalmopathy and several other terms) is an orbital disease associated with thyroid dysfunction. TAO is divided into two types. Active TAO, which usually lasts for 1 to 3 years, is characterized by a persistent autoimmune/inflammatory reaction in the orbital soft tissues. Active TAO causes expansion and remodeling of the ocular soft tissues. The autoimmune/inflammatory reaction of active TAO resolves spontaneously and the condition transforms to inactive TAO. Inactive TAO is a term used to describe the long-term/permanent sequelae of active TAO. The cause of TAO is unknown. TAO is usually associated with Graves' hyperthyroidism, but can also occur as part of other autoimmune conditions that affect the thyroid gland and produce pathology in the orbital and periorbital tissues and rarely in the pretibial skin (pretibial myxedema) or fingers (thyroid clubbing). TAO is an autoimmune orbital disease in which the orbit and periocular soft tissues are primarily affected, with secondary effects on the eyeball and vision. In TAO, inflammation and dilation of the orbital soft tissues (primarily ocular muscles and fat) forces the eyeball forward (bulges) out of its socket - a phenomenon known as proptosis or exophthalmos. Although most cases of TAO do not cause vision loss, the condition can cause vision-threatening exposure keratopathy, troublesome diplopia (double vision), and compressive dysthyroid optic neuropathy. TAO may precede, coincide with, or follow systemic complications of thyroid dysfunction. Ocular clinical manifestations of TAO include upper eyelid retraction, eyelid lag, swelling, redness (erythema), conjunctivitis and bulging eyes (proptosis or exophthalmos), chemosis, periorbital edema, and altered ocular motility, with significant functional, social, and cosmetic consequences. Many of the signs and symptoms of TAO, including bulging eyes and ocular hyperemia, result from expansion of the orbital fat tissue and periocular muscles. The increase in adipose tissue volume is due in part to the development of new adipocytes (lipogenesis) within the orbital fat. Accumulation of hydrophilic glycosaminoglycans, primarily hyaluronic acid, within the perimysial connective tissue between the orbital fat tissue and the extraocular muscle fibers further expands the fat compartment and enlarges the extraocular muscle body. Hyaluronic acid is produced by fibroblasts residing within the orbital fat and extraocular muscles, and its synthesis in vitro is stimulated by several cytokines and growth factors, including IL-1β, interferon-γ, platelet-derived growth factor, thyroid-stimulating hormone (TSH), and insulin-like growth factor I (IGF-I).

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

IGF-IR is a ubiquitously expressed heterotetrameric protein involved in the regulation of proliferation and metabolic functions of many cell types. It is a tyrosine kinase receptor containing two subunits. IGF-IRα contains the ligand binding domain, while IGF-IRβ is involved in signal transduction and contains tyrosine phosphorylation sites.

Current treatments for hyperthyroidism caused by Graves' disease are not perfect because there is a lack of treatments that target the specific underlying pathogenic autoimmune mechanisms of the disease. Even more complicated 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 problem. There are no drugs approved 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 rarely satisfactory. Partial responses are frequent, and relapses (rebounds) are common after discontinuation of the drug. Adverse events occur and many patients eventually need rehabilitation surgery when their condition has been converted to inactive TAO. Therefore, there is still a need to provide alternative therapies for TAO and its related symptoms.

Summary of the invention

The 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 the biological function of IGF-1R signaling mediated by IGF-I. In addition, the present invention generally relates to methods for treating thyroid associated eye disease (TAO), which is also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmos, thyroid dysfunction eye disease and other thyroid eye diseases associated with IGF-1R signaling.

In some embodiments, an antibody or antigen-binding fragment thereof comprising a sequence as provided herein is provided. In some embodiments, the antibody comprises a VL sequence as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86; and a VH sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83. In some embodiments, the antibody comprises a 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, and a 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; or any combination or variant thereof.

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

In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 20, 26, 32, 38, 44, 50 or 56; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 21, 27, 33, 39, 45, 51 or 57; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47 or 53; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 18, 24, 26, 32, 38, 44, 50 or 56; 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 one of the foregoing.

In some embodiments, the antibody or antibody fragment comprises: (i) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 20; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 21; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 22; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 17; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 26; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 27; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 28; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 23; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 24; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 32; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 33; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 34; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 29; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 30; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO:44; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO:45; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO:46; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO:41; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO:42; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 50; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 51; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 52; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 47; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 48; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 56; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 57; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 58; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 53; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 54; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO:62; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO:63; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO:64; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO:59; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO:60; and the light chain CDR3 sequence has an 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 40; or a variant of any of the foregoing; and (ii) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 35; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 81; or a variant of any of the foregoing.

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

In some embodiments, the antibody comprises the sequence of SEQ ID NO: 65 to 72, 78, 82, or 85, or a variant thereof.

In some embodiments, the antibody comprises a light chain having an amino acid sequence of SEQ ID NO: 3 and a heavy chain having an amino acid sequence of SEQ ID NO: 83. In some embodiments, the antibody comprises a light chain variable region having an amino acid sequence of SEQ ID NO: 13 and a heavy chain variable region having an 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 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 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 amino acid sequence of SEQ ID NO:95.

In some embodiments, variants of any of the antibodies provided herein are provided, so 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 described in SEQ ID NO:75 to 77, 84, 87, 88, 89 or 90. In some embodiments, the Fc region is as described in SEQ ID NO:75. In some embodiments, the Fc region is as described in SEQ ID NO:76. In some embodiments, the Fc region is as described in SEQ ID NO:77. In some embodiments, the Fc region is as described in SEQ ID NO:84. In some embodiments, the Fc region is as described in SEQ ID NO:87. In some embodiments, the Fc region is as described in SEQ ID NO:88. In some embodiments, the Fc region is as described in SEQ ID NO:89. In some embodiments, the Fc region is as described in SEQ ID NO:90.

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

In some embodiments, provided is 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 an antibody as provided herein or a pharmaceutical composition comprising the same.

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

In some embodiments, provided is a method of reducing a clinical activity score (CAS) of thyroid associated 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, a method is provided for reducing a) proptosis by at least 2 mm and b) clinical activity score (CAS) in a subject with thyroid associated 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, a method of treating or lessening the severity of thyroid associated eye disease (TAO) in a subject is provided, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces proptosis in one eye by at least 2 mm; (ii) is not accompanied by a deterioration of the other eye (or the fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

In some embodiments, a method of improving the quality of life of a subject with thyroid associated eye disease (TAO, also known as Graves' ophthalmopathy/Graves' orbitopathy) is provided, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

In some embodiments, a method of treating or reducing the severity of diplopia in a subject with thyroid associated eye disease (TAO) is provided, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same.

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

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

In some embodiments, a method of treating thyroid eye disease in a subject is provided, the method comprising administering to the subject an antibody as provided herein, or a pharmaceutical composition comprising the same, wherein at least 1, 2, or 3 weeks after administration, 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.

In some embodiments, a method is provided for inhibiting IGF-1-induced receptor autophosphorylation in a cell by at least 95%, 96%, 97%, 98%, 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 of any one of the methods provided herein are provided, wherein the antibody or its antigen-binding fragment is administered in the form of a pharmaceutical composition that further comprises 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 or other anti-CD20 antibodies; tocilizumab or other anti-IL-6 antibodies; or selenium, infliximab or other anti-TNFα antibodies or thyroid stimulating hormone receptor (TSHR) inhibitors.

BRIEF DESCRIPTION OF THE DRAWINGS

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

FIG. 2 illustrates 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. The antibodies can be used, for example, to treat thyroid eye disease.

As used herein, "thyroid associated eye disease" (TAO), "thyroid eye disease" (TED), "Graves' ophthalmopathy" or "Graves' orbitopathy" (GO) refer to the same disorder or condition and are used interchangeably. They all refer to inflammatory orbital pathology associated with some autoimmune thyroid disorders, most commonly "Graves' disease" (GD), but sometimes with other diseases (e.g., Hashimoto's thyroiditis).

The terms "proptosis" and "exophthalmos" (also known as exophthalmos/exophthalmia/exorbitism) refer to a forward protrusion, displacement, bulging, or protrusion of an organ. As used herein, the terms refer to a forward protrusion, displacement, bulging, or protrusion of the eye forward away from the orbit. Some technicians in the field consider proptosis and exophthalmos to have the same meaning and often use them interchangeably, while others believe that there is a subtle difference in their meanings. Some people use proptosis to refer to severe bulging eyes; or to refer to endocrine-related bulging eyes. Others use the term proptosis when describing bulging eyes associated with, for example, the eyes of subjects with TAO (TED or GO).

As used herein, the terms "exophthalmos" and "proptosis" are used interchangeably and refer to a forward protrusion, displacement, bulging or protrusion of the eye forward away from the orbit. Due to the rigid bony structure of the orbit with only an anterior opening for expansion, any increase in the soft tissue contents of the orbit, either laterally or posteriorly, will displace the eyeball forward. Exophthalmos or proptosis may be the result of several disease processes, including infection, inflammation, tumors, trauma, cancer metastasis, endocrine disorders, vascular disease, and extraorbital lesions. TAO (TED or GO) is currently recognized as the most common cause of exophthalmos in adults. Protrusion may be bilateral, as is common in TAO (TED or GO); or unilateral, as is common in orbital tumors.

The degree of proptosis can be measured using, for example, an exophthalmosometer (an instrument for measuring the degree of forward displacement of the eye). The device allows the measurement of the forward distance from the outer edge of the orbit to the front of the cornea. Computed tomography (CT) scanning and magnetic resonance imaging (MRI) can also be used to assess the degree of proptosis or bulging eyes. CT scanning is an excellent imaging modality for diagnosing TAO. In addition to allowing observation of enlarged extraocular muscles, CT scanning also provides surgeons or clinicians with drawings of the bony anatomy of the orbit when orbital decompression is required. MRI provides excellent imaging of the orbital contents through its multi-plane and inherent contrast capabilities without the need for radiation exposure associated with CT scanning studies. MRI provides better imaging of the optic nerve, orbital fat, and extraocular muscles, but CT scanning provides a better view of the bony architecture of the orbit. Orbital ultrasound examination can also be used to diagnose and assess TAO because it can be performed quickly and with a high degree of confidence. It is easy to assess the hyperreflectivity and enlargement of the extraocular muscles, and continuous ultrasound examinations can also be used to assess the progression or stability of eye disease. Based on technology that is currently available or that will become available in the future, one skilled in the art will be able to determine the best modality for diagnosing and assessing the degree of proptosis or exophthalmos.

As used herein, the term "antibody" refers to any form of an antibody that exhibits the desired biological activity. It is therefore used in the broadest sense and specifically encompasses, 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 before modifying the antibody for the intended use (e.g., humanization of an 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, that is, an antibody fragment that retains the ability to specifically bind to an antigen bound by a full-length antibody, for example, a fragment that retains one or more CDR regions. Examples of antibody binding fragments include, but are not limited to, Fab, Fab', F(ab') ₂ , and Fv fragments; bispecific antibodies; linear antibodies; single-chain antibody molecules, such as sc-Fv; nanobodies and multispecific antibodies formed from antibody fragments.

A "Fab fragment" contains one light chain and the _CH1 and variable region of one heavy chain. The heavy chain of a Fab molecule cannot form disulfide bonds with another heavy chain molecule.

The "Fc" region contains two heavy chain fragments comprising the _CH1 and _CH2 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and hydrophobic interactions of the _CH3 domains.

In some embodiments, the antibody or antigen fragment herein comprises an Fc region. In some embodiments, the Fc region comprises a mutation that prolongs the half-life of the antibody when bonded to the Fc region. In some embodiments, the Fc region comprises S228P, L235E, M252Y, S254T, T256E, M428L, N434S, L234F, P331S mutations or any combination thereof. In some embodiments, the Fc region comprises M252Y, S254T and T256E mutations. Non-limiting examples of Fc regions comprising M252Y, S254T and T256E mutations (collectively referred to as "YTE mutations") can be found in the sequence of SEQ ID NO:89. In some embodiments, the Fc region comprising a YTE mutation comprises a sequence of SEQ ID NO:90, which is different from SEQ ID NO:89 due to the presence of a C-terminal lysine (K) residue. The numbering of the Fc region can be carried out 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 antibody comprises L234F, L235E and P331S mutations. In some embodiments, the Fc region comprises M252Y, S254T, T256E, S228P and L235E mutations. In some embodiments, the Fc region comprises 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, M252Y, 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 Apr; 34(6):441-52, each of which is incorporated herein by reference in its entirety. The numbering cited 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:

A "Fab'fragment" contains one light chain and a portion or fragment of one heavy chain containing the _VH domain and the _CH1 domain and the region between the _CH1 and _CH2 domains, such that an interchain disulfide bond can form between the two heavy chains of the two Fab' fragments to form a F(ab') ₂ molecule.

A "F(ab') ₂ fragment" contains two light chains and two heavy chains containing a portion of the constant region between ^the _CH1 and _CH2 domains, such that an interchain disulfide bond is formed between the two heavy chains. Thus, a F(ab') ₂ fragment is composed of two Fab' fragments held together by a disulfide bond between the two heavy chains.

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

The term "single-chain Fv" or "scFv" antibody refers to an antibody fragment comprising the _VH and _VL domains of an antibody, wherein these domains are present in a single polypeptide chain. In general, the Fv polypeptide further comprises a polypeptide linker between the _VH and _VL domains that enables the scFv to form a desired structure for antigen binding. For a review of scFv, see Pluckthun (1994) PHARMACOLOGY OF MONOCLONAL ANTIBODIES, Vol. 113, Rosenburg and Moore, ed. Springer-Verlag, New York, pp. 269-315. See also International Patent Application Publication No. WO 88/01649 and U.S. Patent Nos. 4,946,778 and 5,260,203.

"Domain antibodies" are immunologically functional immunoglobulin fragments that contain only the variable region of the heavy chain or the variable region of the light chain. In some cases, two or more _VH regions are covalently joined with a peptide linker to produce a bivalent domain antibody. The two _VH regions of a bivalent domain antibody can 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, a bivalent antibody can be bispecific (see below).

In certain embodiments, the 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. Pat. No. 6,005,079. In one embodiment, the invention provides a single domain antibody comprising two _{VH domains} having modifications such that a single domain antibody is formed.

As used herein, the term "bispecific antibody" refers to a small antibody fragment with two antigen-binding sites, which fragment comprises a heavy chain variable domain (VH) connected to a light chain variable domain ( _VL ) in the same polypeptide chain ( _VH - _VL or _VL - _VH ). By using a linker that is too short to allow pairing between the _two domains on the same chain, the domains are forced to pair with the complementary domains of another chain, creating two antigen-binding sites. Bispecific antibodies are more fully described in, e.g., EP 404,097; WO 93/11161; and Holliger et al., (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448. For a review of engineered antibody variants, see generally Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

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

"Isolated antibody" refers to the purified state of the binding compound, which in this context means 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 medium. In general, the term "isolated" is not intended to refer to the complete absence of such materials or the absence of water, buffers, or salts, unless their content substantially interferes with the experimental or therapeutic use of the binding compound as described herein.

As used herein, the term "monoclonal antibody" refers to a substantially homogeneous antibody population, that is, except for possible naturally occurring mutations that may be present in small amounts, the amino acid sequences of the antibody molecules constituting the population are consistent. In contrast, conventional (polyclonal) antibody preparations typically include numerous different antibodies with different amino acid sequences in their variable domains, particularly in their CDRs, and the amino acid sequences are typically specific for different epitopes. The modifier "monoclonal" indicates that the antibody is characterized by being obtained from a substantially homogeneous antibody population, and should not be understood as requiring the antibody to be produced by any particular method. For example, the monoclonal antibodies used according to the present invention can be made by the hybridoma method first described by Kohler et al. (1975) Nature 256:495 or can be made by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). "Monoclonal antibodies" can also be separated from phage antibody libraries using techniques such as 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 domains are obtained from antibodies from experimental animals such as rodents ("parent antibodies"), and the constant domain sequences are obtained from human antibodies, 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 an antibody form containing sequences from human and non-human (e.g., mouse, rat) antibodies. In general, a humanized antibody will comprise substantially all of at least one and usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to those regions of non-human immunoglobulins, and all or substantially all of the framework (FR) regions are those regions of human immunoglobulin sequences. A 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 that contains only human immunoglobulin sequences. If produced in a mouse, a mouse cell, or a hybridoma derived from a mouse cell, a fully human antibody may contain mouse carbohydrate chains. Similarly, a "mouse antibody" refers to an antibody that contains only mouse immunoglobulin sequences. Alternatively, if produced in a rat, a rat cell, or a hybridoma derived from a rat cell, a fully human antibody may contain rat carbohydrate chains. Similarly, a "rat antibody" refers to an antibody that contains only rat immunoglobulin sequences.

Typically, the basic antibody structural unit comprises a tetramer. Each tetramer includes two pairs of identical polypeptide chains, each pair having a "light" chain (about 25kDa) and a "heavy" chain (about 50-70kDa). The amino terminal portion of each chain includes a variable region with about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The carboxyl terminal portion of the heavy chain can define a constant region that is primarily responsible for effector function. Typically, human light chains are classified as κ light chains and λ light chains. In addition, human heavy chains are typically classified as μ, δ, γ, α or ε, and the 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 connected by a "J" region of about 12 or more amino acids, wherein the heavy chain also includes a "D" region with about more than 10 amino acids. See generally Fundamental Immunology Chapter 7 (Paul, W., ed., 2nd ed. Raven Press, NY (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 contain three hypervariable regions, also known as complementarity determining regions (CDRs), which are located within relatively conserved framework regions (FRs). The CDRs are typically aligned through the framework regions to enable binding to specific epitopes. In general, from N-terminus to C-terminus, the light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The assignment of amino acids to domains is generally based on the definitions in Sequences of Proteins of Immunological Interest , Kabat et al.; National Institutes of Health, Bethesda, Md.; 5th 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 (CDRL1), 50-56 (CDRL2), and 89-97 (CDRL3) in the light chain variable domain and residues 31-35 (CDRH1), 50-65 (CDRH2), and 95-102 (CDRH3) in the heavy chain variable domain; Kabat et al., (1991) Sequences of Proteins of Immunological Interest, 5th edition Public Health Service, National Institutes of Health. of Health, Bethesda, Md.) and/or those residues from the "hypervariable loops" (i.e., residues 26-32 (CDRL1), 50-52 (CDRL2), and 91-96 (CDRL3) in the light chain variable domain and 26-32 (CDRH1), 53-55 (CDRH2), and 96-101 (CDRH3) in the heavy chain variable domain; Chothia and Lesk (1987) J. Mol. Biol. 196:901-917). As used herein, the term "framework" or "FR" residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues. The CDRs provide most of the contact residues for antibody binding to an antigen or epitope. The 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 neutralization ability as the donor antibody from which they were derived.

In some embodiments, the antibody may be in the form of a full-length antibody, a single domain antibody, a recombinant heavy chain antibody only (VHH), a single chain antibody (scFv), a shark heavy chain antibody only (VNAR), a microprotein (cysteine knot protein, knottin), ankyrin repeat protein (DARPin); Tetranectin; Affibody; Transbody; Anticalin; AdNectin; Affilin; Microbody; Peptide aptamer; alterase; Plastic antibody; Phylomer ); stradobody; maxibody; evibody; fynomer, armadillo repeat protein, Kunitz domain, avimer, atrimer, probody, immunobody, triomab, troybody; pepbody; vaccibody, unibody; affimer, duobody, Fv, Fab, Fab', F(ab')2, peptide mimetic molecule or synthetic molecule, as disclosed in U.S. Patents or Patent Publication No. US No. 7,417,130, No. US 2004/132094, No. US 5,831,012, No. US2004/023334, No. US 7,250,297, No. US 6,818,418, No. US 2004/209243, No. US 7,838,629, No. US 7,186,524, No. US 6,004,746, No. US 5,475,096, No. US 2004/146938, No. US 2004/157209, No. US 6,994,982, No. US 6,794,144, No. US 2010/239633, No. US No. 7,803,907, No. US 2010/119446 and/or No. US 7,166,697, the contents of each of which are incorporated herein by reference in their entirety. See also Storz MAbs. 2011 May-June; 3(3):310-317, which is incorporated herein by reference.

As used herein, the term "antigen" means any molecule that is capable of directly or indirectly producing an antibody or binding to an antibody. Included within the definition of "antigen" are nucleic acids encoding proteins. "Antigen" may also refer to a binding partner of an antibody. In some embodiments, the antigen is an IGF-1R protein expressed on the surface of a cell. In some embodiments, the cell is an intact cell. An intact cell is a cell that has not been dissolved or broken when a detergent or other reagent is used. Cells that have been treated with detergents or other reagents that destroy the cell membrane or make holes in the cell membrane are not intact cells. For example, provided herein is a method for producing an antibody that binds to an IGF-1R protein, the method comprising culturing a cell comprising a nucleic acid molecule encoding an IGF-1R antibody.

As used herein, "specific binding" or "immunospecific binding" or "immunospecifically binds" 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 binds with a dissociation constant ( _KD ) of ^10-7 M or less, and the _{KD for binding to the predetermined antigen is at least two times less than its KD for} binding to a nonspecific antigen other than the predetermined antigen (e.g., BSA, casein, or another nonspecific polypeptide). The phrases "antibody that recognizes IGF-1R" and "antibody that is specific for IGF-1R" may be used interchangeably herein with the term "antibody that immunospecifically binds to IGF-1R." The present invention may refer 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, the antibodies or binding compounds derived from the antigen binding sites of the antibodies in the contemplated methods bind to their antigen (IGF-1R) with an affinity that is at least two times, at least ten times, at least 20 times, or at least 100 times greater than that of 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 with a sequence homology or consistency between 40% and 100% with a reference sequence. The percent consistency between the two peptide chains can be determined by comparing the default settings of the AlignX module using Vector NTIv.9.0.0 (InvitrogenCorp., Carslbad, Calif.). In some embodiments, the antibody or its antigen-binding fragment has at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homology or consistency with the sequence described herein. In some embodiments, the antibody has a conservative substitution compared to the sequence described herein. Exemplary conservative substitutions are illustrated in Table 1 and are encompassed within the scope of the disclosed subject matter. Conservative substitutions may be present in the framework region or in the antigen binding site, as long as it does not adversely affect the properties of the antibody. Substitutions may be made to improve antibody properties, such as stability or affinity. Conservative substitutions will produce molecules with functional and chemical characteristics similar to those to which such modifications were made. Exemplary amino acid substitutions are shown in the table below.

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

As used herein, the term "in combination with" means that the agents described may be administered to an animal or subject together in a mixture, simultaneously as a single dose, or sequentially in any order as a single dose.

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

The term "epitope" means a portion of any molecule that can be recognized and bound by an antibody at one or more of the antigen binding regions of the 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 characteristics as well as specific charge characteristics. Examples of epitopes include, but are not limited to, the residues that form IGF-1R epitopes described herein. In some embodiments, the epitope is present only in non-denatured proteins. In some embodiments, the epitope is present only in denatured proteins.

In some embodiments, the source of DNA encoding non-human antibodies includes antibody-producing cell lines, such as hybrid cell lines generally known as hybridomas.

Hybrid cells are formed by the fusion of non-human antibody-producing cells (usually spleen cells of animals immunized with natural or recombinant antigens, or peptide fragments of antigenic protein sequences). Alternatively, the non-human antibody-producing cells can be B lymphocytes obtained from the blood, spleen, lymph nodes or other tissues of animals immunized with antigens.

The second fusion partner that provides immortality function can be a lymphoblastoid or plasmacytoma or myeloma cell, which is not a cell that produces antibodies itself, but is malignant. Fusion partner cells include, but are not limited to, hybridoma SP2/0-Ag14, abbreviated as SP2/0 (ATCC CRL1581) and myeloma P3X63Ag8 (ATCC TIB9) or derivatives thereof. See, for example, Ausubel, below, Harlow, below, and Colligan, below, the contents of which are incorporated herein by reference in their entirety.

Antibodies can be produced according to the embodiments provided herein. Once the sequence is known, antibodies can also be produced according to known methods. Antibodies can also be converted into different types, such as human IgG, etc. By converting antibodies into human antibodies, human subjects should not recognize the antibodies as foreign antibodies. It is well known that non-human IgG antibodies are converted into human IgG antibodies, and once the natural sequence is known, the conversion can be performed routinely. 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 therapy. Nature 332 (6162): 332-323; Tsurushita N, Park M, Pakabunto K, Ong K, Avdalovic A, Fu H, Jia A, Vásquez M, Kumar S. (2004). Antibody-producing cells that contribute to the nucleotide sequence encoding the antigen-binding region of the chimeric antibody can also be produced by transforming non-human (such as primate) or human cells. For example, antibody-producing B lymphocytes can be infected and transformed with a virus such as Epstein-Barr virus to obtain immortal antibody-producing cells (Kozbor et al., Immunol. Today 4:72 79 (1983)). Alternatively, B lymphocytes can be transformed by providing a transforming gene or a transforming gene product, as is well known in the art. See, for example, Ausubel below, Harlow below, and Colligan below, 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 art of immunology. Fusion partner cell lines and methods for fusion and selection of hybridomas and screening of mAbs are well known in the art. See, for example, Ausubel below, Harlow below, and Colligan below, 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 amino acids of an epitope of IGF-1R.

In some embodiments, the antibodies comprise a sequence as provided herein.

The sequence of the antibody can be modified to produce human IgG antibodies. The conversion of the sequence provided herein can be modified to produce other types of antibodies. The CDR can also be connected to other antibodies, proteins, or molecules to produce antibody fragments that bind to IGF-1R. It can 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 into fully human according to known methods. The sequence can also be made into chimeric antibodies as described herein.

In some embodiments, the antibody comprises an amino acid sequence or a fragment thereof comprising a sequence provided herein. In some embodiments, the antibody comprises one or more amino acid sequences, its antigen-binding fragment or its human IgG variant as provided herein. "Its human IgG variant" 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 with known sequences is conventional and can be carried out by any method. Therefore, in some embodiments, nucleic acids encoding antibodies or fragments thereof are provided. In some embodiments, nucleic acids encode sequences provided herein. Antibodies may also be modified to chimeric antibodies or human antibodies. Antibodies may also be used in the form of injectable pharmaceutical compositions. Also as described herein, antibodies may be isolated antibodies or engineered antibodies.

In some embodiments, "derivatives" of antibodies, fragments, regions or derivatives thereof are provided, the term including those proteins encoded by genes that are truncated or modified to produce molecular species that are functionally similar to immunoglobulin fragments. 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.

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

The nucleic acid sequence encoding the antibodies described herein may be a genomic DNA or cDNA encoding at least one of the variable regions described herein, or an RNA (e.g., mRNA). A suitable alternative to using chromosomal gene fragments as a DNA source for encoding V region antigen-binding fragments is to use cDNAs for constructing chimeric immunoglobulin genes, such as reported by Liu et al. (Proc. Natl. Acad. Sci., USA 84: 3439 (1987) and J. Immunology 139: 3521 (1987)), which are incorporated herein by reference in their entirety. The use of cDNA requires gene expression elements suitable for host cells to be combined with genes in order to synthesize the desired protein. The use of cDNA sequences is superior to 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 or neutralizing an IGF-1R antigen can 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 can be used to encode a specific amino acid (Watson et al., see below). Using the genetic code, one or more different oligonucleotides can be identified, each of which will be able to encode 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 the eukaryotic or prokaryotic cells expressing the antibody or fragment and the frequency with which a particular codon (encoding a specific amino acid) is actually used. Lathe et al., J. Molec. Biol. 183: 112 (1985) discloses such "codon usage rules". Using Lathe's "codon usage rules", a single oligonucleotide or a set of oligonucleotides containing the theoretically "most likely" nucleotide sequence capable of encoding an antibody variable or constant region sequence is identified.

The variable regions described herein can be combined with any type of constant region (including human constant regions or mouse constant regions). Human genes encoding constant (C) regions, fragments and regions of antibodies can be derived from human fetal liver libraries by known methods. Human C region genes can be derived from any human cell, including human cells that express and produce human immunoglobulins. Human _CH regions can be derived from any of the known classes or isotypes of human H chains, including γ, μ, α, δ or ε, and subtypes thereof, such as G1, G2, G3 and G4. Because the H chain isotype is responsible for the various effector functions of antibodies, the selection of _CH regions will be guided by the desired effector function (such as complement fixation) or activity in antibody-dependent cellular cytotoxicity (ADCC). Preferably, _CH regions are derived from γ1 (IgG1), γ3 (IgG3), γ4 (IgG4) or μ (IgM). Human _CL regions can be derived from human L chain isotypes, κ or λ. In some embodiments, the antibody comprises an Fc domain. In some embodiments, the Fc domain comprises mutations that extend the half-life of the antibody. In some embodiments, the Fc domain comprises mutations, such as those described in U.S. Patent No. 7,670,600, which is incorporated herein by reference 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, and the position is numbered according to the EU numbering index of Kabat. Without being bound by any particular theory, an antibody comprising a mutation corresponding to residue 428 may have an extended half-life compared to the half-life of an IgG with a wild-type human IgG constant domain. In some embodiments, mutations are substituted with threonine, leucine, phenylalanine or serine for a natural residue. In some embodiments, the antibody further comprises one or more amino acid substitutions relative to a corresponding wild-type human IgG constant domain at one or more of amino acid residues 251-256, 285-290, 308-314, 385-389 and 429-436, and the 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 human immunoglobulin C regions can be obtained from human cells by standard cloning techniques (Sambrook et al. (Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Press, Cold Spring Harbor, NY) (1989) and Ausubel et al., ed. Current Protocols in Molecular Biology (1987 1993)). Human C region genes are easily obtained from known clones containing genes representing two categories of L chains, five categories of H chains, and subcategories thereof. Chimeric antibody fragments such as F(ab') ₂ and Fab can be prepared by designing appropriately truncated chimeric H chain genes. For example, a chimeric gene encoding the H chain portion of a F(ab') ₂ fragment will include a DNA sequence encoding the CH ₁ domain and hinge region of the H chain, followed by a translation stop codon, thereby obtaining a truncated molecule.

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

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

Thus, cDNA encoding antibody V and C regions, methods of producing antibodies according to some embodiments described herein, involve several steps, as exemplified below: 1. Isolating messenger RNA (mRNA) from a cell line producing an anti-IGF-1R antigen antibody and from optional additional antibodies supplying heavy and light chain constant regions; cloning and producing cDNA therefrom; 2. Preparing a full-length cDNA library from the purified mRNA, in which suitable V and/or C region gene segments of the L and H chain genes can be: (i) identified with suitable probes, (ii) sequenced and (iii) made compatible with the C or V gene segment of another antibody from a chimeric antibody; 3. Constructing a complete H or L chain coding sequence by ligating the cloned specific V region gene segment to the cloned C region gene as described above; 4. Expressing and producing the L and H chains in a selected host (including prokaryotic and eukaryotic cells) to provide mouse-mouse, human-mouse, human-human or human-mouse antibodies.

Two coding DNA sequences are said to be "operably linked" if the linkage results in a continuous translatable sequence without changes or interruptions in the triple reading frame. A coding sequence is operably linked to a gene expression element if the linkage results in the proper functioning of the gene expression element to result in 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 antibodies described herein are used to detect the presence of an antigen.Antibodies of the invention can be used in any device or method to detect the presence of an antigen.

The term "purified" in reference to an antibody refers to an antibody that is substantially free of other materials associated with molecules in its natural environment. For example, a purified protein is substantially free of cellular material or other proteins of the cells or tissues from which it is derived. The term refers to preparations in which the isolated protein is sufficient to be analyzed 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. 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 to the polypeptides. Techniques and commercially available kits for generating and screening phage display libraries are well known to those skilled in the art. In addition, examples of methods and reagents particularly suitable for generating and screening antibody or antigen binding protein display libraries 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 may also be conjugated to a chemical moiety. The chemical moiety may be, in particular, 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 with a molecular weight of 2kDa, 5kDa, 10kDa, 12kDa, 20kDa, 30kDa, or 40kDa), dextran, and monomethoxypolyethylene 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 (diethylenetriaminepentaacetic acid (DTPA)). Examples of chemical moieties include, but are not limited to, anti-mitotic agents such as calicheamicin (e.g., ozogamicin), monomethyl auristatin E, mertansine, and the like. Other examples include, but are not limited to, biologically active anti-microtubule agents, alkylating agents, and DNA minor groove binders. Other examples are provided herein and below. The chemical moiety can be attached to the antibody via a linker (maleimide), a cleavable linker (e.g., a cathepsin cleavable linker (valine-citrulline)), and in some embodiments one or more spacers (e.g., p-aminobenzylcarbamate). Without being bound by any particular theory, once the antibody conjugate binds to IGF-1R, it can be internalized 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 to 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 ⁵⁶ Fe.

Antibodies and antibody fragments may also be conjugated to fluorescent or chemiluminescent labels, including fluorophores, such as rare earth chelators, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanates, phycoerythrin, phycocyanin, allophycocyanin, o-phthalaldehyde, fluorescamine, ¹⁵² Eu, dansyl, umbelliferone, luciferin, luminal label, isoluminal label, aromatic acridinium ester label, imidazole label, acridimium salt label, oxalate label, aequorin label, 2,3-dihydrophthalazinedione, biotin/avidin, spin labels, and stable free radicals.

The antibody molecules can also be conjugated to cytotoxic factors such as diphtheria toxin, Pseudomonas aeruginosa exotoxin A chain, ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins and compounds (e.g., fatty acids), dianthus proteins, Phytoiacca americana proteins PAPI, PAPII and PAP-S, momordica charantia inhibitor, curcin, crotonin, saponaria officinalis inhibitor, mitogens, restrictocins, phenomycins and enomycins.

Any method known in the art for conjugating the antibody molecules of the invention to various moieties may be employed, 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 for conjugating antibodies are routine and well known in the art.

Chimeric Antigen Receptor

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

In some embodiments, antibodies (e.g., anti-IGF-1R antibodies) are provided herein. In some embodiments, the antibody is a recombinant antibody that binds to an 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 conformation (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 is not naturally occurring. 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 having the following sequence or variants thereof:

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

The columns indicated as antibody sequences include the VH and VL chains of the antibody. Where the 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, the antibody may include the VH and VL sequences provided in the tables provided herein. For example, in some embodiments, the antibody includes one or more VH, HC, LC or VL (those sequences with constant domains are complete light or heavy chains) or variants thereof having the following sequences:

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:

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

As provided herein, the heavy chain can be linked to an Fc region, including those with mutations that can 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 illustrated with VH and VL domains with or without constant regions. Constant regions can be replaced as provided herein. VH regions and VL regions can be used to form antibodies as provided herein. VH and VL sequences can be in any form, including, but not limited to, scFv forms in which the VH and VL regions are connected by a peptide linker. Examples of peptide linkers that can be used to connect 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 connected 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. 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 an 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 a heavy chain or light chain CDR having a sequence 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 a sequence of 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. In some embodiments, the antibody or antibody binding fragment thereof comprises a heavy chain CDR having a sequence of 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.

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

Different CDR motifs can be combined in any combination, including those combinations not depicted in the above table. For example, the following embodiments are provided as non-limiting examples of such combinations.

In some embodiments, the antibody or antigen-binding fragment thereof comprises: (i) a light chain variable region comprising a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 17; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 19; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 20; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 21; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 23; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 24; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 25; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 26; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 27; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 29; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 30; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 31; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 32; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 33; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 35; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 37; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO:41; the light chain CDR2 has an amino acid sequence of SEQ ID NO:42; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO:43; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO:44; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO:45; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 47; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 48; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 49; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 50; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 51; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 53; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 54; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 55; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 56; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 57; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 59; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 60; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 61; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 62; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 63; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 35; the light chain CDR2 has an amino acid sequence of SEQ ID NO: 36; and the light chain CDR3 sequence has an amino acid sequence of SEQ ID NO: 81; and (ii) a heavy chain variable region comprising a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 38; the heavy chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 39; and the heavy chain CDR3 sequence has an 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 by any one of other light chain CDR1 sequences. In some embodiments, the light chain variable region CDR2 is replaced by any one of other light chain CDR2 sequences. In some embodiments, the light chain variable region CDR3 is replaced by any one of other light chain CDR3 sequences. In some embodiments, the heavy chain variable region CDR1 is replaced by any one of other light chain CDR1 sequences. In some embodiments, the heavy chain variable region CDR2 is replaced by any one of other light chain CDR2 sequences. In some embodiments, the heavy chain variable region CDR3 is replaced by any one of other light chain CDR3 sequences.

In some embodiments, an antibody or antigen-binding fragment thereof or a protein is provided that comprises a peptide having a sequence as set forth in any one of SEQ ID NO: 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 its antigen-binding fragment comprises the sequence of SEQ ID NO:65, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:66, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:67, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:68, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:69, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:70, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:71, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:72, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment comprises the sequence of SEQ ID NO:78, or a variant of any of the foregoing. In some embodiments, the antibody or its antigen-binding fragment 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, the _VL and/or _VH sequences are as provided herein. In some embodiments, _{the VL sequence is provided as an element of a light chain (LC). In some embodiments, the VL sequence provided as an element of a light chain (LC) is underlined in the LC sequence. In some embodiments, the VH sequence provided} as an element of a heavy chain (LC) is underlined in the HC sequence.

In some embodiments, the antibody or antigen-binding fragment thereof comprises a _VL peptide as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79, or 86, or any combination thereof. The _VL peptide may comprise a variant of any of these sequences as provided herein.

In some embodiments, the antibody or antigen-binding fragment thereof comprises a _{VH peptide as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80, or 83, or any combination thereof. The VH peptide} may comprise a variant of any of these sequences as provided herein.

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

In some embodiments, the antibody or antigen-binding fragment thereof comprises a LC peptide as set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11, or any combination thereof. The LC peptide may comprise a variant of any of these sequences as provided herein.

In some embodiments, the antibody or antigen-binding fragment thereof comprises a 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 its antigen-binding fragment comprises a HC peptide and a 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 its antigen-binding fragment comprises a HC peptide and a 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 its antigen-binding fragment comprises a HC peptide and a 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, the HC peptide comprising a sequence as set forth in SEQ ID NO: 4 has an additional C-terminal lysine (K) residue. In some embodiments, the antibody or its antigen-binding fragment comprises a HC peptide and a 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 a HC peptide and a 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 a HC peptide and a 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 a HC peptide and a 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 a HC peptide and a LC peptide, wherein the HC peptide comprises a sequence as set forth in SEQ ID NO: 83 and the LC peptide comprises a sequence as set forth in SEQ ID NO: 3.

In addition to these specific combinations, either of the _VH peptide and the _VL peptide may be combined with each other.

In addition to these specific combinations, any of the HC peptides and LC peptides 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 sequences produced by ATCC clone PTA-7444 are incorporated herein by reference in their entirety, including antigen-binding fragments thereof.

Additionally, as provided herein, an antibody can be a multispecific antibody in that the antibody has multiple binding regions that target 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 ( _VH or _VL ) may or may not be connected by a peptide linker and are alternatively contiguous sequences. In some embodiments, the peptide linker comprises the following sequence: (GGGGS) _n (SEQID 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 the formula of _VH - _ZVL or _VL - _ZVH , wherein Z is a peptide linker. In some embodiments, Z is (GGGGS) _n (SEQID NO: 73); (GGGGA) _n (SEQ ID NO: 74), or any combination thereof, wherein each n is independently 1 to 5.

As provided herein, the antibodies or antigen-binding fragments thereof may be sequence variants.

Other examples of antibodies include, but are not limited to, those provided in US20160096894A1, EP1399483B1, EP2194067B1, US20040202651A1, US20110229933A1, US8137933B2, US8951790B2, US20190270820A1, US7572897B2, US20090275126A1, EP1959014B1, US20080014203A1, US20080226635A1, US20120076778A1, US20190153071A1, WO2011161119A1, US106 11825B2, US20120237507A1, EP2681240B1, US9982036B2, US20180312573A1, EP2681239B1, US20160151487A1, US20190225696A1, WO2017011773A2, US20200023076A1, US20190153471A1, US20190194713A1, WO2020006486A1, US20080112888A1, US20150168424A1, EP2032989B2, US9045536B2, each of which is incorporated herein by reference in its entirety. Other examples of antibodies include, but are not limited to, those provided in US8153121B2, EP1469879B1, WO2016064716A1, US20190270820A1, US20180280527A1, US20190225696A1, US7998681B2, US20040202651A1, US2005013606 3A1, US20090285824A1, US20150274829A1, EP2322550B1, US20060286103A1, US20070071675A1, US20100047239A1, US20130004416A1, US20080112888A1, US2 0150168424A1、US20100143 340A1, US20110014117A1, US20100260668A1, US20100074900A1, US20150017168A1, US20110044980A1, US20130330323A1, US20120263722A1, US201202017 46A1、US10519245B2、US2018 0243432A1, US20170218091A1, US20200115460A1, US20100104645A1, US20120065380A1, EP2970433B1, US20160289341A1, US20160289343A1, US20190293656A1, 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 contains the following sequence:

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

And the light chain contains the following sequence:

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

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

Pharmaceutical composition

In some embodiments, to prepare a pharmaceutical composition or sterile composition of an anti-IGF-1R antibody or other protein provided herein, the antibody or antigen-binding fragment thereof or other protein provided herein is blended 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 can 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, e.g., Hardman et al., (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weine et al. (eds.) r and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY). In some embodiments, the antibody is diluted to a suitable concentration in sodium acetate solution pH 5 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.

The toxicity and therapeutic efficacy of the antibody composition administered alone or in combination with another agent can be determined by standard pharmaceutical procedures in cell culture or experimental animals, such as determining _LD50 (the dose that is lethal to 50% of the population) and _ED50 (the dose that is therapeutically effective to 50% of the population). The dose ratio between toxic effects and therapeutic effects is the therapeutic index ( _LD50 / _ED50 ). In certain aspects, antibodies that exhibit a high therapeutic index are desirable. The data obtained from these cell culture assays and animal studies can be used to formulate a range of doses for humans. The dosage of such compounds is preferably within a range of circulating concentrations, including an _ED50 with little or no toxicity. The dosage may vary within this range depending on the dosage form and route of administration employed.

In some embodiments, the compositions of the invention are administered to a subject in accordance with the Physicians' Desk Reference 2003 (Thomson Healthcare; 57th Edition (November 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, dermal, transdermal or intraarterial.

In some embodiments, the antibody or its antigen-binding fragment can be administered by an invasive route such as injection. In some embodiments, the antibody or its antigen-binding fragment or its pharmaceutical composition is administered intravenously, subcutaneously, intramuscularly, intraarterially, intraarticularly (e.g., in an arthritic joint), or by inhalation, aerosol delivery. Administration by non-invasive routes (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 can be administered directly to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retro-orbital sinus. In some embodiments, administration is by injection to the eye, the anterior chamber of the eye, the vitreous chamber of the eye, the suprachoroidal space, or the retro-orbital sinus. In some embodiments, the injection is an intravitreal injection, an intraorbital injection, a retro-orbital injection, an suprachoroidal injection, or an intracameral injection. In some embodiments, the injection is an intravitreal injection. In some embodiments, the injection is an intraorbital injection. In some embodiments, the injection is a retro-orbital injection. In some embodiments, the injection is an suprachoroidal injection. In some embodiments, the injection is an intracameral injection.

In some embodiments, an 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 used to treat thyroid eye disease or a condition associated therewith. Examples of such treatments and therapeutic agents include, but are not limited to, antithyroid drugs, diabetes drugs, beta-blockers, propylthiouracil, methimazole, propranolol, atenolol, metoprolol, nadolol, corticosteroids, metformin, sulfonylureas, meglitinide, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, regular insulin, insulin aspart, insulin glulisine, insulin lispro, insulin protamine, insulin isophane), insulin degludec, insulin detemir, insulin glargine, acarbose, miglitol, acebutolol, atenolol, betaxolol, bisoprolol, carteol, carvedilol, esmolol, labetalol, metoprolol, nadolol, nebivolol, penbutolol, pindolol, propranolol lol), sotalol, timolol, tomolol ophthalmic solution, sitagliptin, saxagliptin, linagliptin, alogliptin, dulaglutide, exenatide, semaglutide, liraglutide, lixisenatide, canagliflozin, dapagliflozin, empagliflozin, or any combination thereof.

The compositions can be administered by medical devices known in the art.For example, the pharmaceutical compositions of the invention can be administered by injection with a hypodermic needle (including, for example, a prefilled syringe or an autoinjector).

The pharmaceutical composition may also be administered using a needle-free subcutaneous injection device; such as the devices disclosed in U.S. Pat. Nos. 6,620,135; 6,096,002; 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824 or 4,596,556.

The pharmaceutical composition can also be administered by infusion. Examples of well-known implants and module forms for administering pharmaceutical compositions include: U.S. Pat. No. 4,487,603, which discloses an implantable microinfusion pump that distributes drugs at a controlled rate; U.S. Pat. No. 4,447,233, which discloses a drug infusion pump that delivers drugs at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses an implantable infusion device with a variable flow rate for continuous drug delivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drug delivery system with a multi-chamber compartment. Many other such implants, delivery systems, and modules are well known to those skilled in the art.

Alternatively, the antibody can be administered in a local rather than systemic manner, for example, by direct injection of the antibody into an arthritic joint or pathogen-induced lesion characterized by immunopathology, usually in the form of a reservoir or sustained release formulation. In addition, the antibody can 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, an arthritic joint or pathogen-induced lesion characterized by immunopathology. The liposomes will target the diseased tissue and be selectively absorbed by the diseased tissue.

The administration regimen depends on several factors, including the serum or tissue conversion 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 enough therapeutic antibodies to achieve an improvement in the target disease condition while minimizing undesirable side effects. Therefore, the amount of the biological preparation delivered depends in part on the severity of the specific therapeutic antibody and the condition being treated. Guidance for selecting appropriate doses of therapeutic antibodies is available (see, e.g., Wawrzynczak (1996) Antibody Therapy, Bios Scientific Pub. Ltd, Oxfordshire, UK; Kresina (ed.) (1991) Monoclonal Antibodies, Cytokines and Arthritis, Marcel Dekker, New York, NY; Bach (ed.) (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; Milgrom et al. (1999) New Engl. J. Med. 341:1966-1973; Slamon et al. (2001) New Engl. J. Med. 344:783-792; Beniaminovitz et al. (2000) New Engl. J. Med. 342:613-619; Ghosh et al. (2003) New Engl. J. Med. 348:24-32; Lipsky et al. (2000) New Engl. J. Med. 343:1594-1602).

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

The antibody or its antigen-binding fragment can be provided by continuous infusion or by, for example, a dose administered once a day, 1 to 7 times a week, once a week, once every two weeks, once a month, once every two months, once a quarter, once every six months, once a year, etc. The dose can be provided, for example, intravenously, subcutaneously, topically, orally, nasally, rectally, intramuscularly, intracranially, intraspinally, 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 for at least 21 weeks on this schedule. In some embodiments, the antibody is administered for at least 24 weeks on this schedule. In some embodiments, the antibody is administered for at least 32 weeks on this schedule. In some embodiments, the antibody is administered for at least 36 weeks on this schedule. In some embodiments, the antibody is administered for at least 40 weeks on this schedule. In some embodiments, the antibody is administered for at least 42 weeks on this schedule. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) once. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) twice. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) three times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) four times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) five times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) six times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) seven times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) eight times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) nine times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 10 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 11 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 12 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 13 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 14 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 15 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 16 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 17 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 18 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 19 times. In some embodiments, the antibody is administered (e.g., infusion or subcutaneous injection) 20 times. When the antibody is administered more than once, the antibody may be administered according to a schedule such as the schedule provided herein.

The total weekly dosage may be provided as herein. In some embodiments, the total weekly dose is at least 0.05 μg/kg body weight, more typically at least 0.2 μg/kg, 0.5 μg/kg, 1 μg/kg, 10 μg/kg, 100 μg/kg, 0.25 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 5.0 mg/ml, 10 mg/kg, 25 mg/kg, 50 mg/kg or more (see, e.g., Yang et al. (2003) New Engl. J. Med. 349:427-434; Herold 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). Doses can also be provided to achieve a predetermined target concentration of the antibody in the subject's serum, 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 for at least 1, 2 or 3 weeks after administration.

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

As used herein, "inhibit" or "treat" or "treatment" includes delaying the development of symptoms associated with a disorder and/or lessening the severity of symptoms of such a disorder. The terms further include ameliorating existing uncontrolled or unwanted symptoms, preventing additional symptoms, and ameliorating or preventing the underlying cause of such symptoms. Thus, the terms indicate that a beneficial result has been conferred on a vertebrate subject having a disorder, disease or symptom or at risk of developing such a disorder, disease or symptom.

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 the progression of such a disease or condition when administered to a cell, tissue or subject alone or in combination with an additional therapeutic agent. A therapeutically effective dose further refers to an amount of a binding compound sufficient to cause at least a partial improvement in symptoms, such as to treat, cure, prevent or improve a related medical condition or to increase the rate of treating, curing, preventing or improving such a condition. When applied to individual active ingredients administered alone, a therapeutically effective dose refers only to that ingredient. When applied to a combination, a therapeutically effective dose refers to the combined amount of active ingredients that produces a therapeutic effect, whether administered in combination, serially or simultaneously. An effective amount of a therapeutic agent will improve a diagnostic measure or parameter by at least 10%; typically by at least 20%; preferably by at least about 30%; more preferably by at least 40%, and most preferably by at least 50%. In cases where a subjective measure is used to assess the severity of a disease, an effective amount may also cause an improvement in the subjective measure. In some embodiments, an amount is a therapeutically effective amount if it is an amount that can be used to treat or ameliorate a condition as provided herein.

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

Given that the isolated antibodies bind to an epitope on an IGF-1R protein or other protein described herein, and exhibit in vitro and/or in vivo IGF-1R inhibitory or therapeutic activity, the antibodies or antigen-binding fragments thereof capable of inhibiting IGF-1R function are suitable for use as therapeutic agents for treating IGF-1R-related conditions in humans and animals. These conditions include thyroid eye disease. Thus, methods of treating such conditions are also provided, wherein the methods comprise administering an antibody or antigen-binding fragment thereof to a subject suffering from such a condition.

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

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

In some embodiments, the antibody used is compatible with the recipient species such that an immune response against the MAb does not produce an unacceptably short circulating 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 as described herein. The antibody 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 agent, 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, the dose of the administered agent will vary depending on factors such as the patient's age, weight, height, sex, general medical condition, past medical history, etc.

Antibodies capable of treating conditions associated with IGF-1R activity or for treating IGF-1R-associated pathologies are intended to be provided to a subject in an amount sufficient to affect a reduction, regression or improvement of IGF-1R-associated symptoms or pathologies. Such pathologies include thyroid eye disease, among others.

Thus, in some embodiments, methods of treating a subject suffering from an IGF-1R mediated disorder are provided. In some embodiments, the method comprises administering a pharmaceutical composition comprising an antibody or an antigen binding fragment thereof as provided herein. In some embodiments, the disorder is thyroid eye disease. As provided herein, the antibody or an antigen binding fragment 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 can be used to treat thyroid eye disease. In some embodiments, the antibodies or antigen-binding fragments thereof can be used to treat thyroid associated eye disease (TAO) or its symptoms or to reduce its severity.

In some embodiments, methods or uses are provided for reducing proptosis in an eye of a subject suffering from thyroid associated eye disease (TAO).

In some embodiments, the subject is one that has been previously treated with an antibody other than those provided herein.

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

In some embodiments, methods or uses are provided for a) reducing proptosis in an eye of a subject with thyroid associated eye disease (TAO) by at least 2 mm and b) reducing the clinical activity score (CAS).

As used herein, the term Clinical Activity Score (CAS) refers to the scheme described and scored according to Table 2. According to this scheme, one point is given for each parameter evaluated in the following table. The sum of all points defines the clinical activity and provides the CAS, where 0 or 1 constitutes inactive disease and 7 is severely active eye disease.

As provided in Table 2, CAS consists of seven components: spontaneous retrobulbar pain, pain when trying to move the eye (gaze up, side to side, and down), conjunctival redness, eyelid redness, chemosis, caruncle/fold swelling, and eyelid swelling. Each component is scored as present (1 point) or absent (0 points). 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 inactive disease and 7 is severe active eye disease. A change of >2 points is considered clinically significant.

Item 1, spontaneous orbital pain can be a feeling of pain or pressure on or behind the globe. This pain can be caused by increased intra-orbital pressure when the orbital tissue volume increases through excessive synthesis of extracellular matrix, fluid accumulation, and cell infiltration and proliferation. Item 2, gaze-induced orbital pain can be eye pain when looking or trying to look up, down, or sideways, that is, pain when the eyes move up, down, or sideways or when trying to move the eyes. This type of pain can be caused by stretching of inflamed muscles, especially when trying to gaze upward. ‘Stretching pain’ is not caused by pressing the fingers on the eyeball, which would be expected if it was a manifestation of increased intra-orbital pressure. Both types of pain can be reduced after anti-inflammatory treatment. Therefore, these types of pain are considered to be directly related to autoimmune inflammation of the orbit and are therefore suitable for evaluating TAO activity.

Swelling in TAO is considered as item 6 chemosis (edema of the conjunctiva) in Table 1, and swelling of the caruncle and/or plica semilunaris. Both are signs of active TAO. Eyelid swelling can be caused by edema, fat prolapse or fibrotic degeneration of the entire orbital septum. In addition to swelling, other symptoms indicating active TAO include redness and/or pain of the conjunctiva, eyelids, caruncle and/or plica semilunaris.

In some embodiments, the proptosis of the treated subject is reduced by at least 2 mm. In some embodiments, the proptosis of the treated subject is reduced by at least 3 mm. In some embodiments, the proptosis of the treated subject is reduced by at least 4 mm.

In some embodiments, in the treated subject, 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) is reduced to zero (0).

In some embodiments, a method of treating or lessening the severity of thyroid associated eye disease (TAO) in a subject is provided, wherein treatment with the antibody (i) reduces proptosis in one eye by at least 2 mm; (ii) is not associated with a deterioration of the other eye (or fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

In some embodiments, methods are provided for improving the quality of life of a subject with thyroid associated eye disease (TAO, also known as Graves' ophthalmopathy/Graves' orbital disease). In some embodiments, the quality of life is measured by the Graves' ophthalmopathy quality of life (GO-QoL) assessment or its visual function or appearance subscale. In some embodiments, treatment improves the GO-QoL by greater than or equal to 8 points. In some embodiments, treatment improves the functional subscale of the GO-QoL. In some embodiments, treatment improves the appearance subscale of the GO-QoL.

In some embodiments, a method of treating diplopia or reducing the severity of diplopia in a subject with thyroid associated eye disease (TAO) is provided. In some embodiments, the diplopia is constant diplopia. In some embodiments, the diplopia is non-constant diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, after stopping the administration of the antibody, the improvement of diplopia or the reduction of its severity continues for at least 20 weeks. In some embodiments, after stopping the administration of the antibody, the improvement of diplopia or the reduction of its severity continues for at least 50 weeks.

The severity of the disease can be measured in the following non-limiting embodiments. For example, for the eyelid aperture, the distance between the eyelid edges is measured (in mm) when the patient is in the first eye position, sitting and relaxed and looking at the distance. For the swelling of the eyelid, the measurement/assessment is "absent/unclear", "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, inflammation of the lacrimal caruncle or fold is absent or present. The same Hertel exophthalmos meter and the same canthus distance are used to measure proptosis for individual patients, in millimeters. Subjective diplopia is scored from 0 to 3 (0 = no diplopia; 1 = intermittent, that is, diplopia in the first gaze position when tired or when waking up for the first time; 2 = non-constancy, that is, diplopia during extreme gaze; 3 = constancy, that is, continuous diplopia in the first eye position or reading position). For eye muscle involvement, monocular movement (duction) is measured, expressed in degrees. Corneal involvement is absent/punctate or corneal lesions/ulcers. For optic nerve involvement, that is, best corrected visual acuity, color vision, optic disc, relative pupillary afferent defect, the condition is absent or present. In addition, when optic nerve compression is suspected, the visual field is tested. In some embodiments, patients can be classified according to the following severity classifications. For example, vision-threatening thyroid eye disease: patients with dysthyroid optic neuropathy (DON) and/or corneal rupture. This category requires immediate intervention. Moderate to severe thyroid eye disease: patients without vision-threatening disease, but whose eye disease is sufficient to affect daily life to justify the risk of immunosuppression (if active) or surgical intervention (if inactive). Patients with moderate to severe thyroid eye disease usually have any one or more of the following: eyelid retraction greater than or equal to 2 mm, moderate or severe soft tissue involvement, proptosis that is more than 3 mm higher or 3 mm higher than the normal value for race and gender, non-constant or constant diplopia. Mild thyroid eye disease: Patients whose features of thyroid eye disease have only a minor impact on daily life and are not sufficient to justify immunosuppression or surgical treatment. They usually have only one or more of the following: mild eyelid retraction (<2 mm), mild soft tissue involvement, proptosis <3 mm above normal for their race and sex, transient diplopia or absence of diplopia, and corneal response to lubricant exposure.

In some embodiments, patients can be characterized by a Graves' ophthalmopathy quality of life (GO-QoL) score. In addition to proptosis (or exophthalmos) and CAS, the GO quality of life (GO-QoL) questionnaire is also used to assess quality of life. This questionnaire is designed to measure the improved quality of life after treatment with the methods disclosed herein. In some embodiments, the questionnaire can measure the side effects after treatment with antibodies or antigen-binding fragments thereof according to the methods disclosed herein are reduced or absent compared to treatment with glucocorticoids. GO-QoL is a 16-item self-administered questionnaire divided into 2 subsets and is used to assess the subject's perceived impact of TED on (i) their daily physical activities related to visual function and (ii) psychosocial function. Quality of life is assessed using the GO QoL questionnaire. The GO-QoL questionnaire [C.B.Terwee et al., 1998] is completed at day 1 and week 6, week 12 and week 24 (or PW) during the treatment period and at month 7 and month 12 (or PW) during the follow-up period. GO-QoL is a 16-item self-administered questionnaire divided into two self-assessment subscales; one subscale covers the impact of visual function on daily activities and the other subscale assesses the impact of self-perceived appearance. The visual function subscale covers activities such as driving, walking outdoors, reading, and watching television. The appearance subscale asks subjects questions such as whether the eye disease changes the subject's appearance, causes others to have negative reactions to the subject, causes social isolation, and causes the subject to try to hide their appearance. Each subscale has 8 questions, answered with yes - very much so; yes - a little; or no - not at all. Each question is scored from 0 to 2, and the total raw score is then mathematically converted to a 0-100 scale, where 0 represents the greatest negative impact on quality of life and 100 represents no impact. A change of > or equal to 8 points on the 0-100 scale is shown to be clinically meaningful. The combined score takes the raw scores from both subscales and converts them back to a single 0-100 scale. The questionnaire has two self-assessment subscales. Each subscale has 8 questions answered with (i) yes - very much so; (ii) yes - a little; or (iii) no - not at all. Each question is scored from 0 to 2, and the total raw score is then mathematically converted to a 0-100 scale, where 0 represents the greatest negative impact on quality of life and 100 represents no impact. A change of >8 points on the 0-100 scale is considered clinically meaningful. The combined score takes the raw scores from both subscales and converts them into a single 0-100 scale.

Patients can also be assessed by the presence or absence of the Gorman grade of diplopia. The Gorman assessment of subjective diplopia includes four categories: no diplopia (absent), diplopia when the patient is tired or awake (intermittent), diplopia at extremes of gaze (non-constant), and continuous diplopia in the primary or reading position (constant). Patients are scored according to the grade of diplopia they experience. An improvement of greater than or equal to 1 grade is considered clinically meaningful.

In some embodiments, the method includes administering an antibody, such as those provided herein. In some embodiments, the antibody is administered as a first dose at a dosage of about 1 mg/kg to about 5 mg/kg of the antibody. In some embodiments, the antibody is administered as a first dose at a dosage of about 5 mg/kg to about 10 mg/kg of the antibody. In some embodiments, the antibody is administered as a subsequent dose at a dosage of about 5 mg/kg to about 20 mg/kg of the antibody. In some embodiments, the antibody is administered in the following amounts: about 10 mg/kg of the antibody as a first dose; and about 20 mg/kg of the antibody as a subsequent dose. 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-CD20 antibodies; tocilizumab 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 include administering to a subject an antibody or antigen-binding fragment thereof that specifically binds to and inhibits IGF-IR. In some embodiments, the antibody is as provided herein.

Also provided are kits that can be used to implement the embodiments described herein. The kit of the present invention includes a first container containing the antibody described above or co-packaged with the antibody. The kit may also include another container containing a solution necessary or suitable for implementing the embodiment or co-packaged with the solution. The container may be made of glass, plastic or foil, and may be a vial, bottle, pouch, tube, bag, etc. The kit may also contain written information, such as a procedure for implementing the embodiment; or analytical information, such as the amount of reagent contained in the first container member. The container may be located in another container device (e.g., a box or bag) together with the written information.

Another aspect provided herein is a kit for detecting an IGF-1R protein in a biological sample. The kit includes a container containing one or more antibodies that bind to an epitope of an IGF-1R protein and instructions for using the antibodies for the following purposes: allowing the antibodies to bind to the IGF-1R protein to form an immune complex and detecting the formation of the immune complex so that the presence or absence of the immune complex is associated 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 antibodies are separated. In some embodiments, the antibodies specifically bind. In some embodiments, the antibodies bind to properly folded IGF-1R proteins. In some embodiments, the antibodies are specific to specific IGF-1R conformational states (open or closed). In some embodiments, the antibodies bind to IGF-1R proteins in cell membranes. In some embodiments, the antibodies bind to IGF-1R proteins in cell membranes in intact cells. In some embodiments, the antibodies inhibit or neutralize the function of IGF-1R proteins. As used herein, the term "neutralize" means that the activity or function of the protein is inhibited. The 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 inhibition percentage may be based on the function or activity of the protein in the absence of the antibody. In some embodiments, the antibodies inhibit glucose transport promoted by IGF-1R. In some embodiments, the antibody inhibits internalization of the 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 or an antigen binding fragment thereof as described herein. The heavy chain may be one or more of the heavy chains described herein. In some embodiments, the antibody comprises a light chain or an antigen binding fragment thereof as described herein.

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

In some embodiments, a method of detecting the presence or absence of IGF-1R in a sample is provided, the method comprising contacting the sample with one or more antibodies described herein, thereby detecting binding of the antibody to the IGF-1R antigen. In some embodiments, detection of binding indicates the presence of the IGF-1R antigen; or failure to detect binding to the IGF-1R antigen indicates the absence of the IGF-1R antigen. Detection can be performed by any known method, such as using a biosensor, ELISA, sandwich assay, etc. However, in some embodiments, the method comprises detecting the presence of the protein in non-denaturing conditions. Non-denaturing conditions can 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 a test antibody with an epitope on an 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 the 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 the epitope or protein and determining binding of the test antibody to the mutated epitope, wherein if the mutation reduces binding of the test antibody compared to the non-mutated epitope, then the test antibody is considered to bind to that epitope.

In some embodiments, a method for monitoring IGF-1R internalization from a cell surface is provided. In some embodiments, the method comprises contacting a cell with an anti-IGF-1R antibody as provided herein and detecting the presence of IGF-1R in the cell 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 (such as a test agent) regulating the internalization of an IGF-1R protein. Therefore, the antibodies provided herein can be used to identify a test agent that regulates (increases or decreases) the internalization of an IGF-1R protein. A test molecule that increases internalization can be identified according to the methods provided herein, and the increase in internalization will be measured as a decrease in the binding of an anti-IGF-1R antibody to an IGF-1R protein on the cell surface. A test molecule that reduces internalization can be identified according to the methods provided herein, and the decrease in internalization will be measured as an increase in the binding of an anti-IGF-1R antibody to an IGF-1R protein on the cell surface. Surface expression can be measured by fluorescence, which can be performed by a secondary antibody that recognizes an IGF-1R antibody or by labeling an anti-IGF-1R antibody provided herein.

In some embodiments, a method of inhibiting IGF-1 stimulated receptor phosphorylation on a cell is provided. In some embodiments, the method comprises contacting a cell with an antibody as provided herein or a pharmaceutical composition comprising the same. In some embodiments, contacting comprises administering the 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 suffers from thyroid eye disease (TED) or is at risk thereof. In some embodiments, the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09nm. In some embodiments, the IC50 is measured in vitro, as measured in an assay as provided herein, as described in the examples. In some embodiments, the IC50 is measured in a cell, and the cell is an A549 cell or a HOCF cell.

In some embodiments, a method for treating thyroid eye disease in a subject is provided, the method comprising administering to the subject an antibody as provided herein or a pharmaceutical composition comprising the same, wherein at least 1, 2, or 3 weeks after administration, 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. In some embodiments, the serum concentration is measured after one, two, or three doses of the antibody or a pharmaceutical composition comprising the same are administered to the subject.

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

Implementation options listed

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

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

A VL sequence as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86;

A VH sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83;

A 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

A 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 variation 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 a scFv antibody.

6. The antibody of any one of embodiments 1 to 5, wherein the antibody or antigen-binding fragment thereof comprises a _VL peptide as set forth in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 79 or 86, or any variant thereof.

7. The antibody of any one of embodiments 1 to 6, wherein the antibody or antigen-binding fragment thereof comprises a _VH peptide as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83, 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 a heavy chain CDR1, CDR2 and CDR3 sequence, wherein the heavy chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 20, 26, 32, 38, 44, 50 or 56; the heavy chain CDR2 has an amino acid sequence of SEQ ID NO: 21, 27, 33, 39, 45, 51 or 57; and the heavy chain CDR3 sequence has an 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 a light chain CDR1, CDR2 and CDR3 sequence, wherein the light chain CDR1 sequence has an amino acid sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47 or 53; the light chain CDR2 sequence has an amino acid sequence of SEQ ID NO: 17, 23, 29, 35, 41, 47 or 53 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 one 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 to 17, wherein the heavy chain variable region and the light chain variable region are not connected by a linker.

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

20. The antibody of embodiment 19, wherein the peptide linker comprises the following 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.

21. The antibody of any one of embodiments 1 to 20, wherein the antibody comprises the sequence of SEQ ID NO: 65 to 72, 78, 82 or 85, or a variant thereof.

22. The antibody of any one of embodiments 1 to 21, wherein the antibody comprises a _VL sequence as set forth in: SEQ ID NO: 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 a _VH sequence as set forth in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 80 or 83, 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 NO: 65 to 72, 78, 82 or 85, or a variant 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 is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% homologous to the sequence of SEQ ID NO: 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 NO: 1 to 72, 78 to 83 or 85 to 86.

31. The antibody of any one of embodiments 1 to 26, wherein the antibody is a 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 of the preceding embodiments, wherein the antibody comprises an Fc region.

35. The antibody of embodiment 34, wherein the Fc region is as described in SEQ ID NO:75 to 77 or 84.

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

37. The antibody of embodiment 36, wherein the Fc region comprises S228P, L235E, M252Y, S254T, T256E, M428L, N434S, L234F, P331S mutations, 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, M428L 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, M252Y, S254T and T256E mutations.

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

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

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

48. A pharmaceutical composition comprising the antibody or a nucleic acid molecule encoding the same as described in any one of embodiments 1 to 44.

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

50. A method of treating or reducing 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 embodiments 1 to 44 or a pharmaceutical composition comprising the same.

51. A method of reducing proptosis in an eye of a subject with 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.

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 a subject's Clinical Activity Score (CAS) for 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.

54. A method of reducing a) proptosis by at least 2 mm and b) clinical activity score (CAS) in a subject with 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 convex eye is reduced by at least 2 mm.

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

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

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

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

61. A method for treating or reducing the severity of thyroid associated eye disease (TAO) in a subject, comprising administering to the subject an antibody as described in any one of embodiments 1 to 44, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces proptosis of one eye by at least 2 mm; (ii) is not accompanied by a deterioration of the other eye (or the fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

62. A method of improving the quality of life of a subject with thyroid associated eye disease (TAO, also known as Graves' ophthalmopathy/Graves' orbitopathy), comprising administering to the subject an antibody as described in 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 the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or its visual function or appearance subscales.

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

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

66. The method of embodiment 63, wherein the treatment improves the appearance subscale of the GO-QoL.

67. A method of treating or reducing the severity of diplopia in a subject with 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.

68. A method as described in embodiment 67, wherein the diplopia is constant diplopia.

69. A method as described in embodiment 67, wherein the diplopia is non-constant diplopia.

70. A method as described in embodiment 67, wherein the diplopia is intermittent diplopia.

71. The method of embodiment 67, wherein the improvement in diplopia or reduction in its severity persists for at least 20 weeks after cessation of antibody administration.

72. The method of embodiment 67, wherein the improvement in diplopia or reduction in its severity persists 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 1 mg/kg to about 5 mg/kg of 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 5 mg/kg to about 10 mg/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 5 mg/kg to about 20 mg/kg of antibody.

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

77. The method of embodiment 76, wherein the subsequent doses are 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, Fab, Fab', F(ab')2, Fv or 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-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab or other anti-TNFα antibody or a thyroid stimulating hormone receptor (TSHR) inhibitor.

82. The method of any 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 retro-orbital 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, retro-orbital, 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 for 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 to 44, or a pharmaceutical composition comprising the same.

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

90. The method of embodiment 89, wherein the subject has or is at risk for 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 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm.

92. A method as described in embodiment 91, wherein the IC50 is measured in an in vitro assay, such as an assay provided herein.

93. The method of any one of embodiments 88 to 92, wherein the cell is an A549 cell or a HOCF cell.

94. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as described in any one of embodiments 1 to 44 or as provided elsewhere herein, or a pharmaceutical composition comprising the same, wherein at least 1, 2 or 3 weeks after administration, 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.

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 for 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 described in any one of embodiments 1 to 44 or as provided elsewhere herein, or a pharmaceutical composition comprising the same.

99. The method of embodiment 98, wherein inhibition of IGF-1 induced receptor autophosphorylation is measured 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 the 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 for 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%, 99% or 100%, the method comprising administering to the subject an antibody as described in 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 for 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 the CDRs of VRDN-1100.

106. The method of any one of embodiments 98 to 104, wherein the antibody comprises the CDRs of the antibody of VRDN-1100 or the CDRs 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 amino acid sequence of SEQ ID NO:95.

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

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

114. A pharmaceutical composition suitable for subcutaneous administration, comprising the antibody of any one of embodiments 107 to 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 to 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 reducing the severity of thyroid associated eye disease (TAO) or a symptom thereof, the method comprising administering to a subject an antibody as described in any one of embodiments 107 to 111 or a pharmaceutical composition comprising the same.

119. A method of reducing proptosis in an eye of a subject with thyroid associated eye disease (TAO), comprising administering to the subject the antibody of any one of embodiments 107 to 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 to 111, or a pharmaceutical composition comprising the same.

121. A method of reducing a subject's clinical activity score (CAS) for thyroid associated eye disease (TAO), comprising administering to the subject an antibody as described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising the same.

122. A method of reducing a) proptosis by at least 2 mm and b) clinical activity score (CAS) in a subject with thyroid associated eye disease (TAO), comprising administering to the subject an antibody as described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising the same.

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

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

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

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

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

129. A method for treating or reducing the severity of thyroid associated eye disease (TAO) in a subject, comprising administering to the subject an antibody as described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces proptosis of one eye by at least 2 mm; (ii) is not accompanied by a deterioration of the other eye (or the fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

130. A method of improving the quality of life of a subject with thyroid associated eye disease (TAO, also known as Graves' ophthalmopathy/Graves' orbitopathy), comprising administering to the subject an antibody as described in any one of embodiments 107 to 111, or a pharmaceutical composition comprising the same.

131. The method of embodiment 130, wherein quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or its visual function or appearance subscales.

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

133. The method of embodiment 130, wherein the treatment results in an improvement in the Functional subscale of the GO-QoL.

134. The method of embodiment 130, wherein the treatment improves the appearance subscale of the GO-QoL.

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

136. A method as described in embodiment 135, wherein the diplopia is constant diplopia.

137. A method as described in embodiment 135, wherein the diplopia is non-constant diplopia.

138. A method as described in embodiment 135, wherein the diplopia is intermittent diplopia.

139. The method of embodiment 135, wherein the improvement in diplopia or reduction in its severity persists for at least 20 weeks after cessation of antibody administration.

140. The method of embodiment 135, wherein the improvement in diplopia or reduction in its severity persists for at least 50 weeks after cessation of antibody administration.

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

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

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

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

145. The method of embodiment 144, wherein the subsequent doses are 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 the form of 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. A method as described in embodiment 146 or 147, wherein the pharmaceutical composition further comprises a corticosteroid; rituximab or other anti-CD20 antibody; tocilizumab or other anti-IL-6 antibody; or selenium, infliximab or other anti-TNFα antibody or thyroid stimulating hormone receptor (TSHR) inhibitor.

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

150. The method of embodiment 149, wherein contacting comprises administering the 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 for 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 to 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 to 44, or a pharmaceutical composition comprising the same.

154. The method of embodiment 153, wherein the subject has or is at risk for 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 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm.

156. A method as described in embodiment 155, wherein the IC50 is measured in an in vitro assay, such as an assay provided herein.

157. The method of any one of embodiments 152 to 157, wherein the cell is an A549 cell or a HOCF cell.

158. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as described in any one of embodiments 107 to 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 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 a subsequent dose at a dose of about 1 mg/kg to about 5 mg/kg (mg antibody/kg subject), about 5 mg/kg to about 10 mg/kg antibody, or about 5 mg/kg to about 20 mg/kg.

161. The method of any one of embodiments 158 to 160, wherein the antibody is administered at: about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg of 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 20 mg/kg of antibody as subsequent doses.

162. The method of any one of embodiments 158 to 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%, 99% or 100%, the method comprising administering to the subject an antibody as described in any one of embodiments 107 to 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, M252Y, S254T and T256E substitutions.

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

168. A pharmaceutical composition as described in embodiment 164, wherein the antibody comprises a light chain having an amino acid sequence of SEQ ID NO:93 and a heavy chain amino acid sequence of SEQ ID NO:92.

169. A pharmaceutical composition as described in embodiment 164, wherein the antibody comprises a light chain having an amino acid sequence of SEQ ID NO:93 and a heavy chain amino acid sequence of SEQ ID NO:94.

170. A pharmaceutical composition as described in embodiment 164, wherein the antibody comprises a light chain having an 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 a pharmaceutical composition comprising an antibody as described in any one of embodiments 164 to 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 reducing the severity of thyroid associated eye disease (TAO) or a symptom thereof in a subject, the method comprising administering to the subject a pharmaceutical composition as described in any one of embodiments 164 to 170.

175. A method of reducing proptosis in an eye of a subject having thyroid associated eye disease (TAO), the method comprising administering to the subject the pharmaceutical composition of any one of embodiments 164 to 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 to 4.

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

178. A method of causing a) a reduction in proptosis by at least 2 mm and b) a reduction in Clinical Activity Score (CAS) in a subject with thyroid associated eye disease (TAO), the method comprising administering to the subject a pharmaceutical composition as described in any one of embodiments 164 to 170.

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

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

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

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 for treating or lessening the severity of thyroid associated eye disease (TAO) in a subject, comprising administering to the subject a pharmaceutical composition as described in any of embodiments 164 to 170, wherein treatment with the antibody (i) reduces proptosis in one eye by at least 2 mm; (ii) is not associated with a deterioration of the other eye (or the fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

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

187. The method of embodiment 186, wherein quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or its visual function or appearance subscales.

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

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

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

191. A method of treating or reducing the severity of diplopia in a subject with thyroid associated eye disease (TAO), the method comprising administering to the subject a pharmaceutical composition as described in any one of embodiments 164 to 170.

192. A method as described in embodiment 191, wherein the diplopia is constant diplopia.

193. A method as described in embodiment 191, wherein the diplopia is non-constant diplopia.

194. A method as described in embodiment 191, wherein the diplopia is intermittent diplopia.

195. The method of embodiment 191, wherein the improvement in diplopia or reduction in its severity persists for at least 20 weeks after cessation of antibody administration.

196. The method of embodiment 191, wherein the improvement in diplopia or reduction in its severity persists 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 a first dose at a dose of about 1 mg/kg to about 5 mg/kg, about 5 mg/kg to about 10 mg/kg, about 10 mg/kg to about 20 mg/kg, about 20 mg/kg to about 30 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, or about 30 mg/kg of the antibody.

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

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 1 mg/kg to about 10 mg/kg, about 2 mg/kg to about 5 mg/kg, or about 5 mg/kg to about 20 mg/kg of the antibody.

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

201. The method of embodiment 200, wherein the subsequent doses are 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 longer.

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

203. The method of embodiment 202, wherein contacting comprises administering to the 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 for thyroid eye disease (TED).

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

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

207. The method of embodiment 206, wherein the subject has or is at risk for 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 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm.

209. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject a pharmaceutical composition as described in any of embodiments 164 to 170, 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 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, S254T 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 M428L and N434S mutations.

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

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

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

217. 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 211 to 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 for treating or reducing the severity of thyroid associated eye disease (TAO) or a symptom thereof, the method comprising administering to a subject an antibody as described in any one of embodiments 211 to 213 or a pharmaceutical composition comprising the same.

221. A method of reducing proptosis in an eye of a subject with thyroid associated eye disease (TAO), comprising administering to the subject an antibody as described in any one of embodiments 211 to 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 to 213, or a pharmaceutical composition comprising the same.

223. A method of reducing a subject's clinical activity score (CAS) for thyroid associated eye disease (TAO), comprising administering to the subject an antibody as described in any one of embodiments 211 to 213, or a pharmaceutical composition comprising the same.

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

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

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

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

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 for treating or reducing the severity of thyroid associated eye disease (TAO) in a subject, comprising administering to the subject an antibody as described in any one of embodiments 1 to 3, or a pharmaceutical composition comprising the same, wherein treatment with the antibody (i) reduces proptosis of one eye by at least 2 mm; (ii) is not accompanied by a deterioration of the other eye (or the fellow eye) by 2 mm or more; and (iii) reduces the subject's CAS to one (1) or zero (0).

232. A method for improving the quality of life of a subject with thyroid associated eye disease (TAO, also known as Graves' ophthalmopathy/Graves' orbitopathy), comprising administering to the subject an antibody as described in 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 the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or its visual function or appearance subscales.

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

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

236. The method of embodiment 232, wherein the treatment improves the appearance subscale of the GO-QoL.

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

238. A method as described in embodiment 237, wherein the diplopia is constant diplopia.

239. A method as described in embodiment 237, wherein the diplopia is non-constant diplopia.

240. A method as described in embodiment 237, wherein the diplopia is intermittent diplopia.

241. The method of embodiment 237, wherein the improvement in diplopia or reduction in its severity persists for at least 20 weeks after cessation of antibody administration.

242. The method of embodiment 237, wherein the improvement in diplopia or reduction in its severity persists for at least 50 weeks after cessation of antibody administration.

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

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

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

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

247. The method of embodiment 246, wherein the subsequent doses are 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 longer.

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

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

250. The method of embodiment 249, wherein the subject has or is at risk for 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 of any one of embodiments 211 to 213, or a pharmaceutical composition comprising the same.

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

253. The method of embodiment 252, wherein the subject has or is at risk for thyroid eye disease (TED).

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

255. A method of treating thyroid eye disease in a subject, the method comprising administering to the subject an antibody as described in any of embodiments 211 to 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 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 is now described with reference to the following examples. These examples are provided for illustrative purposes only, and the claims should in no way be construed as being limited to these examples, but rather should be construed as covering any and all variations that become apparent as a result of the teachings provided herein. Those skilled in the art will readily recognize a variety of non-critical parameters that can be changed or modified to produce essentially similar results.

Example

Example 1: IGF-1R antibody blocks IGF-1 stimulation.

Blockade of IGF-1 stimulation was measured by hyaluronic acid secretion in the presence of IGF-1R antibodies VRDN-2700, VRDN-03100, VRDN-02100, VRDN-02200, VRDN-02300, VRDN-02400, VRDN-02500, VRDN-01100, VRDN-02600, and VRDN-02301, all disclosed herein. Immunoglobulins were purified from sera of patients with Graves' ophthalmopathy (GO) and tested for their ability to directly activate TSHR and/or IGF-1R as well as TSHR/IGF-1R cross talk in primary cultures of GO fibroblasts. Cells were treated with M22 or GO-Ig with or without IGF-1R inhibitory 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. Hyaluronan (HA) secretion was measured as the primary biological response to GO fibroblast stimulation. IGF-1R autophosphorylation was used as a measure of direct IGF-1R activation. TSHR activation was determined by the production of cyclic AMP (cAMP). IGF-1R antibodies as disclosed herein were found to effectively block HA secretion and, therefore, were found to block IGF stimulation.

Example 2: Treatment of patients with thyroid eye disease and clinical evaluation of IGF-1R antibodies for thyroid eye disease.

Subjects are provided with infusions of IGF-1R inhibitory 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. The number of infusions is individualized for each subject and is based on the clinical judgment of the investigator. The Day 1 visit occurs within 14 days of the final visit of the previous trial. The visit window for Weeks 1 and 4 is ±1 day, and for Weeks 3, 6, 9, 12, 15, 18, 21, and 24 is ±3 days. The follow-up period is only for subjects who were proptosis non-responders in the previous trial; subjects who relapsed in the previous trial did not participate in the follow-up period. The consultation window during the follow-up period was ±7 days.

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

Subjects who are bulging eye non-responders are scheduled to participate in the 6-month follow-up period in this extension study; subjects who relapse in the run-in study and are retreated in this extension study will not participate in the follow-up period.

Efficacy will be assessed in both eyes at each assessment time point. The "study eye" (i.e., the more severely affected eye) will remain the same eye identified at the baseline (Day 1) visit of the previous study. Efficacy will be assessed in both eyes, but the primary outcome measure will be assessed using the study eye.

Efficacy was assessed by proptosis (measured as part of the clinical severity measure using the Hertel instrument to ensure consistency of measurement), CAS (7-item scale), diplopia (measured as part of the clinical severity measure), and the clinical severity measure (including assessment of motor limitations).

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

Safety was assessed through AE and concomitant medication use monitoring, immunogenicity testing, physical and ophthalmologic examinations, vital signs, clinical safety laboratory assessments (complete blood count, chemistry (including thyroid panel and HbA1C), and urinalysis), pregnancy testing (if applicable), and electrocardiogram (ECG). The study was also monitored by a Data Safety Monitoring Board (DSMB).

Protrusion assessment was performed using a Hertel exophthalmos meter to ensure consistency of measurement, and the same Hertel instrument and the same observer were used for each assessment for the full duration of the study (unless unavoidable). In addition, the same intercanthal distance (ICD) was used in each case.

Proptosis was measured in each eye at Day 1 and Weeks 6, 12, 18, and 24 (or premature discontinuation (PW)) during the treatment period and Months 7, 9, and 12 (or PW) during the follow-up period. Measurements of eCRF, a clinical severity measure for proptosis, were recorded.

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

Example 3: Antibodies with increased pK

Cynomolgus macaques were given an antibody comprising the CDR of VRDN-2700 with a YTE mutation in the Fc domain at an amount of 10 mg/kg by intravenous or subcutaneous routes, and samples were collected at 0.5 hours, 2 hours, 8 hours and days 1, 3, 7, 10, 14, 21 and 28 for PK analysis by ELISA. Tetumumab was also administered IV at 10 mg/kg as a comparator. The results shown in Figure 1 indicate that the antibody has a significantly higher PK than Tetumumab.

This result suggests that antibodies comprising the CDRs of VRDN-2700 may potentially be given at lower doses compared to tebuconazole, even when administered subcutaneously. These results were not predicted.

Embodiment 4:

VRDN-1100 is an antagonist antibody to the insulin-like growth factor-1 receptor (IGF-1R) being developed for the treatment of thyroid eye disease (TED). TED is driven by agonistic autoantibodies to the thyroid stimulating hormone receptor (TSHR) and crosstalk between TSHR and IGF-1R. TED is characterized by the recruitment of IGF-1R and TSHR-expressing fibrocytes into orbital tissue, where they mediate the deposition of hyaluronic acid and the expansion of orbital muscle and fat1. IGF-1R antagonism was found to reverse this orbital tissue expansion and robustly relieve symptoms in patients with TED2.

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): Anti-Fc capture antibody was immobilized and recombinant IGF-1R extracellular domain (ECD) was flowed as analyte. Association and dissociation rate constants (ka and kd, respectively), as well as the equilibrium dissociation constant, KD, were derived by global fitting of the data to a single-site model.

Epitope binning: VRDN-1100 was immobilized on the chip surface by amine coupling and used to capture IGF-1R-ECD, after which Tetumumab was flowed over the chip.

Cell binding: A549 human lung adenocarcinoma cells or primary human ocular choroidal fibroblasts (HOCF) were incubated with different concentrations of VRDN-1100 or tetumumab. A single dose of 50 nM IgG1 isotype control was used as a 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 live cells. The median fluorescence intensity (MFI) of live cells was measured by flow cytometry and data were analyzed using FlowJo software. Dose curves were fitted using a nonlinear regression model; log (agonist) versus response-variable slope (four parameters).

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

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

Antagonism: Serum starved A549 or HOCF cells were pre-incubated with various concentrations of test antibodies for one hour at 37°C and subsequently stimulated by the addition of 100 ng/mL (A549) or 200 ng/mL (HOCF) IGF-1 for 7 minutes at 37°C. Phosphorylated IGF-1R (pIGF1R) of biological replicates was measured using the R&D Systems pIGF-1R ELISA according to the manufacturer's protocol, and pIGF-1R concentrations were normalized to the lowest test antibody concentration. Dose curves were fitted using a nonlinear regression model; log(inhibitor) versus response - variable slope (four parameters).

result

VRDN-1100 binds to IGF-1R with sub-nanomolar affinity. FIG. 2A shows that increasing concentrations of IGF-1R-ECD bound to anti-FC captured VRDN-1100 or teixumab revealed a stepwise increase in the SPR signal, enabling a global fit to the binding model. After IGF-1R clearance, VRDN-1100 exhibited a more sustained binding interaction. FIG. 2B shows that IGF-1R-ECD is firmly bound to immobilized VRDN-1100. Teixumab did not show binding to the IGF-1R:VRDN-1100 complex, indicating that teixumab and VRDN-1100 have overlapping epitopes. The data are also illustrated in the table shown in FIG. 2 .

VRDN-1100 binds to IGF-1R on A549 cells with high affinity. As shown in FIG3 , the binding of VRDN-1100 to A549 cells was assessed by flow cytometry and was found to have a similar binding profile to teuximab at three different concentrations. Also shown in FIG3 , the binding dose response curve shows that VRDN-1100 EC50 = 0.1 nM. As shown in FIG3 , VRDN-1100, VRDN-2700 with M252Y, S254T, and T256E mutations in the Fc domain, and teuximab exhibit similar binding at temperatures that block IGF-1R receptor internalization. FIGD shows that VRDN-1100, VRDN-2700 with M252Y, S254T, and T256E mutations in the Fc domain, and teuximab cause similar levels of internalization (approximately 50%), as measured by a reduction in membrane IGF-1R receptor levels at 37° C. versus 4° C. In the bar graphs of FIG3 , the leftmost bar graph is the isotype control, the second bar graph set from the left is tetumumab, the second bar graph set from the right is 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 implicated in extracellular matrix deposition and pathogenic fibrosis in TED5. As shown in Figure 4, HOFCs were shown to express (Panel A) IGF-1R and (Panel B) TSHR, as well as (Panel C) CD34 and Thy-1, confirming their ability to serve as an in vitro model system for IGF-1R function in TED.

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

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

VRDN-1100 is a sub-nanomolar IGF-1R antagonist. VRDN-1100 potently inhibits IGF-1 stimulated receptor phosphorylation on A549 cells (IC50 = 0.09 nM) and HOCF cells (IC50 = 0.09 nM), as shown in Figure 6, Panels A and B.

These results demonstrate that the VRDN-1100 and tebuconazole epitopes on IGF-1R overlap, that VRDN-1100 binds to IGF-1R on cells with a subnanomolar EC50, that VRDN-1100 promotes IGF-1R internalization, and that VRDN-1100 inhibits IGF-1R phosphorylation with a subnanomolar IC50. Thus, VRDN-1100 binds, antagonizes, and internalizes IGF-1R at subnanomolar concentrations, suggesting that VRDN-1100 should be used to potentially potently inhibit the pathophysiology that drives TED.

Example 4. VRDN-2700 with M252Y, S254T and T256E mutations in the Fc domain is a novel anti-IGF-1R antibody as described herein, incorporating half-life extension modifications in the Fc region of the antibody, and can be used to treat thyroid eye disease (TED). The pharmacokinetic (PK) parameters of VRDN-2700 with such Fc mutations were measured in cynomolgus monkeys relative to the marketed IGF-1R antibody tetumumab, and a PK model was constructed to plan possible human dosing regimens.

TED is an autoimmune condition most commonly associated with Graves' disease and hyperthyroidism, but can also be seen in euthyroid or hypothyroid patients. Eye disease in TED is driven by thyroid stimulating hormone receptor (TSHR) agonistic autoantibodies and crosstalk between TSHR and IGF-1R. Pathological remodeling of the orbit and periorbital tissues produces different manifestations, which can include dry eyes, increased tearing, local irritation, eyelid retraction and eventually proptosis, diplopia, and optic nerve compression with consequent visual acuity loss.

The underlying pathology of TED is the activation of an inflammatory cascade within the orbit, primarily due to the recruitment of fibrocytes and immune cells. Overexpression of IGF-1R has been demonstrated in the orbit of TED patients, and it has been hypothesized that IGF-1R inhibitory antibodies could disrupt IGF-1R and TSHR crosstalk and attenuate the inflammatory cascade. Indeed, IGF-1R antagonism has been shown to robustly alleviate many of the inflammatory symptoms affecting TED patients.

VRDN-2700 is a monoclonal antibody that inhibits IGF-1-mediated signaling through IGF-1R at sub-nanomolar potency and incorporates clinically validated half-life-extending Fc modifications (M252Y, S254T, and T256E). This antibody was found to have a more favorable PK profile compared to conventional IgG therapeutic antibodies, potentially a less burdensome treatment modality for patients.

VRDN-2700 with Fc mutations was administered to cynomolgus monkeys at 2, 10, and 50 mg/kg by 30 min intravenous (IV) infusion, and at 2 and 10 mg/kg by subcutaneous (SC) injection. 10 mg/kg of titumumab was similarly administered by 30 min IV infusion. Human IgG-specific ELISA was used to measure VRDN-2700 and titumumab levels in serum. Data were analyzed using the WinNonlin non-compartmental model. A semi-mechanistic model incorporating target-mediated drug disposition was constructed using available human and cynomolgus monkey data. The data are described below.

FIG7 is a table and graph illustrating a more favorable PK profile.

The table presents PK parameters +/- SD. Evidence of target mediated drug disposition (TMDD) was observed at 2 mg/kg, but not at 10 and 50 mg/kg doses, consistent with reports of TMDD saturation at higher doses for tefixime and other IGF-1R antibodies.

VRDN-2700 Half-Life Extension Modification Prolongs Exposure

At equivalent doses, SC-administered VRDN-2700 with the YTE mutation had greater exposure than intravenously infused telutumumab and achieved approximately 2-fold the half-life of telutumumab in NHPs, and the bioavailability (F) of SC-administered VRDN-2700 was estimated to be 62% using the initial discovery phase formulation. Parameter estimates +/- SD are shown in FIG8 .

Model simulations predict that administration of VRDN-2700 at 10 mg/kg every 3 weeks or 20 mg/kg every 6 weeks will produce a Cmin of >100 ug/mL, similar to the approved regimen for tebuconazole (10 mg/kg first dose followed by seven 20 mg/kg doses q3w). The 10 mg/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 mitigate toxicity. In addition, the model predicts that a 300 mg fixed dose of VRDN-2700 administered subcutaneously weekly can achieve a steady-state Cmin of approximately 130 ug/mL, enabling self-administration at home. In the case of an effective lower Cmin value, a 300 mg fixed dose of VRDN-2700 administered subcutaneously every other week is predicted to achieve a steady-state Cmin level of approximately 50 ug/mL. In summary, the extended half-life of VRDN-2700 is predicted to provide patients with a wider range of choices for more appropriate dosing intervals and routes of administration.

Example 5: VRDN2700 Properties During the evaluation of the antibodies, the expression of VRDN-2700 was compared to other antibodies with mutations in the Fc domain (such as the L/S mutation described herein). Unexpectedly, the yield of the antibody with the YTE mutation in the Fc domain (VRDN2700) was approximately 80% higher than the yield of the similar antibody with the L/S mutation. This was surprising and unexpected because other antibodies targeting IGF-1R that had been tested with YTE or LS mutations had similar expression regardless of the Fc mutation. The YTE version had fewer low molecular weight species than the LS version. Therefore, it was shown that the YTE antibody had fewer impurities and was a more homogeneous composition, which provided an advantage over antibodies with LS mutations. This was also unpredictable because another antibody evaluated showed the opposite effect on such species. In addition, during purification, it was found that the LS mutant formed more aggregates than VRDN-2700 when purified on a cation exchange column. Aggregation of the LS mutant would cause substantial manufacturing problems, no such problems were observed for VRDN-2700. Thus, this difference in the Fc mutants for this antibody may not have been predicted or anticipated and results in substantial 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 injections, or as an IV infusion, requires fewer and/or less frequent treatments compared to conventional therapeutic IgG antibodies, and has superior properties compared to other Fc mutant versions of the same antibody (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 an Fc YTE mutation in the heavy chain (SEQ ID NO: 94) or a C22S mutation and an 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 had the sequence of SEQ ID NO: 93. The YTE Fc mutant version of VRDN1100 was found to bind to A549 cells with an EC50 of 0.30 nm and the C22S and Fc YTE mutants had an EC50 of 0.36 nm. The ability of the antibodies to inhibit IGF-1R autophosphorylation was also evaluated. The YTE mutant alone had an IC50 of 0.40 nm and the C22S plus YTE mutation had an IC50 of 0.37 nm. Thus, the antibody was found to both bind to IGF-1R and inhibit its autophosphorylation.

Example 7: VRDN-1100 with C22S mutation binds to IGF-1R. Mutants of VRDN-1100 with a C22S mutation in the heavy chain (SEQ ID NO: 96) and a 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 antibody was found to bind to IGF-1R at pH 7.4 with a k _a (1/Ms) of 1.04×10 ⁵ , a k _d (1/s) of 2.18×10 ^-5 , and a K _D (M) of 2.10×10 ^-10 .

Each of these examples and embodiments provided herein demonstrates 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 was specifically and individually indicated as incorporated by reference. Applicants comply with 37 C.F.R. §1.57(b)(1), and this statement of incorporation by reference is with respect to each individual publication, database entry (e.g., Genbank sequence or GeneID entry), patent application, or patent, each of which is expressly identified pursuant to 37 C.F.R. §1.57(b)(2), even if such reference is not adjacent to the specific statement of incorporation by reference. The inclusion of specific statements of incorporation by reference (if any) in this specification does not in any way weaken this general statement of incorporation by reference. Citation of references herein is not intended to admit that the references are relevant background art, nor does it constitute any admission of the contents or dates of these publications or documents.

The scope of the embodiments of the present invention is not limited by the specific embodiments described herein. In fact, various modifications other than those described herein will be apparent to those skilled in the art from the foregoing description. These modifications are intended to fall within the scope of the embodiments and any appended claims.

This description is considered to be sufficient to enable one skilled in the art to practice the embodiments. Various modifications in addition to those shown and described herein will be apparent to those skilled in the art from the foregoing description and fall within the scope of the present disclosure and any appended claims.

Sequence Listing

<110> Viridian Therapeutics, Inc.

<120> Compositions and methods for treating thyroid eye disease

<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

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

210 215

<210> 2

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

1 5 10 15

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

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> 3

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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

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

210 215

<210> 4

<211> 447

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 4

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

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 Lys

435 440 445

<210> 5

<211> 214

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 5

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

<210> 6

<211> 460

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 6

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 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

<211> 214

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 7

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

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 8

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

<211> 219

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 9

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

<212> PRT

<213> Artificial Sequence

<220>

<223> Synthetic sequence

<400> 10

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 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

<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

<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

<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

<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

<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

<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

<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

<220>

<223> Synthetic sequence

<400> 18

Asp Ala Ser Lys Arg Ala Thr

1 5

<210> 19

<211> 10

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 20

Ser Tyr Gly Met His

1 5

<210> 21

<211> 17

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 24

Lys Val Ser Asn Arg Leu Tyr

1 5

<210> 25

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 26

Gly Gly Tyr Leu Trp Asn

1 5

<210> 27

<211> 16

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 30

Gly Glu Asn Lys Arg Pro Ser

1 5

<210> 31

<211> 11

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 32

Ser Tyr Ala Ile Ser

1 5

<210> 33

<211> 17

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 36

Ala Ala Ser Arg Leu His Arg

1 5

<210> 37

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 38

Ser Tyr Ala Met Asn

1 5

<210> 39

<211> 17

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 40

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

1 5 10 15

<210> 41

<211> 16

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 42

Leu Gly Ser Asn Arg Ala

1 5

<210> 43

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 44

Ser Ser Ser Asn Trp Trp Ser

1 5

<210> 45

<211> 16

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 48

Tyr Ala Ser Gln Ser Leu Ser

1 5

<210> 49

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 50

Ser Phe Ala Met His

1 5

<210> 51

<211> 16

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 54

Lys Val Ser Asn Arg Phe Ser

1 5

<210> 55

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 56

Ser Tyr Trp Met His

1 5

<210> 57

<211> 18

<212> PRT

<213> 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

<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

<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

<220>

<223> Synthetic sequence

<400> 60

Asp Ala Ser Ser Leu Gln Thr

1 5

<210> 61

<211> 9

<212> PRT

<213> 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

<220>

<223> Synthetic sequence

<400> 62

Ile Tyr Arg Met Gln

1 5

<210> 63

<211> 16

<212> PRT

<213> 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

<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

<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

<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

<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 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

<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 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

<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 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

<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

<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 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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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 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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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

<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 (46)

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, a variable heavy chain comprising the sequence of SEQ ID NO:99 and an Fc region comprising the mutations M252Y, S254T and T256E. 3. An isolated antibody comprising a variable light chain comprising the sequence of SEQ ID NO:98, 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 1-3. 5. A pharmaceutical composition suitable for intravenous administration comprising the antibody of any one of claims 1-3. 6. A pharmaceutical composition suitable for subcutaneous administration comprising the antibody of any one of claims 1-3. 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 1-3. 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 thyroid-associated ophthalmopathy (TAO) or its symptoms or reducing its severity, said method comprising administering to a subject the antibody of any one of claims 1 to 3 or a medicament comprising it combination. 11. A method of reducing exophthalmos in a subject's eye suffering from thyroid-associated ophthalmopathy (TAO), comprising administering to the subject the antibody of any one of claims 1 to 3 or a medicament comprising it combination. 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 thyroid-associated ophthalmopathy (TAO) in a subject, comprising administering to the subject the antibody of any one of claims 1 to 3, or comprising its pharmaceutical composition. 14. A method of reducing a) exophthalmos by at least 2 mm and b) reducing clinical activity score (CAS) in a subject suffering from thyroid-associated ophthalmopathy (TAO), comprising administering to the subject as claimed in claim 1 The antibody described in any one of to 3, or a pharmaceutical composition comprising it. 15. The method of any one of claims 10 to 14, wherein the protruding eye is reduced by at least 2mm. 16. The method of any one of claims 10 to 14, wherein the protruding eye is reduced by at least 3 mm. 17. The method of any one of claims 10 to 14, wherein the protruding eye is reduced by at least 4 mm. 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 for treating thyroid-associated ophthalmopathy (TAO) in a subject or reducing its severity, comprising administering to the subject the antibody of any one of claims 1 to 3, or a medicament comprising it A composition wherein treatment with said antibody (i) reduces exophthalmos in one eye by at least 2mm; (ii) is not accompanied by deterioration of the other eye (or fellow eye) by 2mm or more; and (iii) causes The subject's CAS is reduced to one (1) or zero (0). 22. A method for improving the quality of life of a subject suffering from thyroid-associated ophthalmopathy (TAO, also known as Graves ophthalmopathy/Graves orbitopathy), said method comprising administering to the subject the The antibody described in any one of to 3, or a pharmaceutical composition comprising it. 23. The method of claim 22, wherein the quality of life is measured by the Graves Ophthalmic Quality of Life (GO-QoL) assessment or its visual function or appearance subscales. 24. The method of claim 22, wherein the treatment results in an improvement in GO-QoL of greater than or equal to 8 points. 25. The method of claim 22, wherein the treatment results in an improvement in a functional subscale of GO-QoL. 26. The method of claim 22, wherein the treatment results in an improvement in the Appearance subscale of GO-QoL. 27. A method of treating or reducing the severity of diplopia in a subject suffering from thyroid-associated ophthalmopathy (TAO), said method comprising administering to the subject the compound of any one of claims 1 to 3 Antibodies, or pharmaceutical compositions comprising them. 28. The method of claim 27, wherein the diplopia is constant diplopia. 29. The method of claim 27, wherein the diplopia is non-constant diplopia. 30. The method of claim 27, wherein the diplopia is intermittent diplopia. 31. The method of claim 27, wherein the improvement in diplopia or reduction in severity thereof persists for at least 20 weeks after cessation of antibody administration. 32. The method of claim 27, wherein the improvement in diplopia or reduction in severity thereof persists 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 present at about 1 mg/kg to about 5 mg/kg, about 5 mg/kg to about 10 mg/kg, about 10 mg/kg to about 20 mg/kg kg, about 20 mg/kg to about 30 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, or about 30 mg/kg of the antibody is administered as the first dose . 34. The method of any one of claims 7-32, wherein the antibody is administered as the first dose at a dose of about 10 mg/kg to about 20 mg/kg antibody. 35. The method of any one of claims 7 to 32, wherein the antibody is present at about 1 mg/kg to about 10 mg/kg, about 2 mg/kg to about 5 mg/kg, or about 5 mg/kg to about 20 mg Doses per kg antibody were administered as subsequent doses. 36. The method of any one of claims 7-32, wherein the antibody is administered in the following amounts: about 10 mg/kg antibody as a first dose; and about 20 mg/kg antibody as a subsequent dose. 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 longer time. 38. A method of increasing IGF-1R internalization on a cell, said method comprising contacting said 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 said contacting comprises administering said antibody or a pharmaceutical composition comprising 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, said method comprising making said cell with the antibody of any one of claims 1 to 3 or a pharmaceutical composition comprising it touch. 42. The method of claim 41, wherein the contacting comprises administering to the subject the antibody of any one of claims 1-3 or a pharmaceutical composition comprising the same. 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 to 43, wherein the antibody has an IC50 of less than or equal to about 0.2nm, 0.15nm, 0.10nm, 0.09nm. 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 it, wherein the administration At least 1, 2 or 3 weeks thereafter, the serum concentration of said antibody in said 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. 46. The method of claim 45, wherein said antibody or said pharmaceutical composition is administered intravenously or subcutaneously.