Composition and method for treating thyroid eye disease

Abstract

The present invention provides an anti-IGF-1R antibody and its composition and uses.

Description

Technology Field

[0001] 관련 출원에 대한 상호 참조

[0002] This application claims the benefit and priority of U.S. Application No. 63 / 619,033 filed January 9, 2024, the full contents of which are incorporated herein by reference. Background Technology

[0003] Thyroid-associated ophthalmopathy (TAO), also known as thyroid eye disease (TED), Graves' ophthalmopathy or orbitopathy (GO), thyrotoxic exophthalmos, dysthyroid ophthalmopathy, and various other terms, is an orbital disorder associated with thyroid dysfunction. TAO is classified into two types. Active TAO, which typically lasts for 1 to 3 years, is characterized by an autoimmune / inflammatory response progressing in the orbital soft tissues. Active TAO is involved in the expansion and remodeling of the ocular soft tissues. The autoimmune / inflammatory response of active TAO resolves spontaneously, and the condition transitions to inactive TAO. Inactive TAO is a term describing the long-term / permanent sequelae of active TAO. Chronic TAO is a stage of inactive TAO and can be characterized in subjects first diagnosed with TAO 2 to 10 years prior to screening, where the Clinical Activity Score (CAS) is 1 or less in both eyes for at least one year prior to screening, or where there is no progression of proptosis, no progression of diplopia, and no new inflammatory TAO symptoms for one year prior to screening. The cause of TAO is unknown. Although TAO is typically associated with Graves' hyperthyroidism, it can also occur as part of other autoimmune conditions affecting the thyroid, causing lesions in the orbit and periorbital tissues, and rarely, may cause lesions in the cutaneous tissue of the anterior tibial bone (anterior tibial myxedema) or fingers and toes (acromegaly). TAO is an autoimmune orbital disease that primarily affects the orbit and periorbital soft tissues, resulting in secondary effects on the eye and vision.In TAO, inflammation and expansion of the orbital soft tissues, primarily the ocular muscles and lipids, cause the eyeball to be pushed forward out of the orbit (protrusion); this condition is referred to as proptosis or exophthalmos. While vision loss does not occur in most cases of TAO, this condition can lead to vision-threatening exposure keratopathy, uncomfortable diplopia (double vision), and compressive dysthyroid optic neuropathy. TAO may appear before, simultaneously with, or after systemic complications of thyroid dysfunction. Ocular symptoms of TAO include upper eyelid retraction, ptosis prolapse, swelling, redness (erythema), conjunctivitis, and protruding eyeballs (proptosis or exophthalmos), conjunctival edema, periorbital edema, and altered ocular motility accompanied by significant functional, social, and cosmetic consequences. Many signs and symptoms of TAO, including proptosis and conjunctival hyperemia, are caused by the expansion of orbital adipose tissue and periorbital muscles. The volume of adipose tissue increases partly due to the development of new adipocytes (lipogenesis) within the orbital fat. The accumulation of hydrophilic glycosaminoglycans, primarily hyaluronic acid, within the periorbital connective tissue between the orbital adipose tissue and the extraocular muscle fibers further expands the fat compartment and hypertrophies the extraocular muscle body. Hyaluronic acid is produced by fibroblasts present in the orbital fat and extraocular muscles, and in vitro, its synthesis is stimulated by several cytokines and growth factors, including IL-1β, interferon-gamma, platelet-derived growth factor, thyroid-stimulating hormone (TSH), and insulin-like growth factor 1 (IGF-1).

[0004] Antibodies activating the insulin-like growth factor 1 receptor (IGF-1R) were also detected in active TAOs and were implicated. While we do not wish to be bound by any theory, TSHR and IGF-1R form physical and functional complexes in orbital fibroblasts, and blocking IGF-1R appears to attenuate both IGF-1-dependent and TSH-dependent signaling. Since blocking IGF-1R using antibody antagonists reduces both TSHR-dependent and IGF-1-dependent signaling, it has been suggested that this may interfere with the pathological activity of autoantibodies acting as agonists at either receptor.

[0005] IGF-1R is a widely expressed heterotemeric protein involved in the regulation of proliferation and metabolic functions in many cell types. It is a tyrosine kinase receptor containing two subunits. IGF-1Rα contains a ligand-binding domain, while IGF-1Rβ is involved in signal transduction and contains a tyrosine phosphorylation site.

[0006] Current therapy for hyperthyroidism due to Graves' disease is incomplete because it lacks therapies that target the specific underlying pathogenic autoimmune mechanisms of the disease. The treatment of moderate to severe active TAO is even more complex. Despite improved understanding of its etiology in recent years, TAO remains a therapeutic challenge and dilemma. There are no approved drugs to treat active TAO. Intravenous glucocorticoids (ivGCs) and oral glucocorticoids are used to treat patients with moderate to severe active TAO, but the results are often unsatisfactory. Partial responses are frequent, and relapse (rebound) after drug discontinuation is not uncommon. Adverse events occur, and many patients eventually require rehabilitation surgery performed after their condition has progressed to inactive TAO. Therefore, there is still a need to provide alternative therapies for TAO and its associated symptoms. means of solving the problem

[0007] The present invention provides an antibody comprising a variable heavy chain having an amino acid sequence that is at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence that is at least 90% identical to SEQ ID NO. 2. In some embodiments, the variable heavy chain comprises glycine corresponding to position 26 of SEQ ID NO. 1, tyrosine corresponding to position 27 of SEQ ID NO. 1, threonine corresponding to position 28 of SEQ ID NO. 1, phenylalanine corresponding to position 29 of SEQ ID NO. 1, threonine corresponding to position 30 of SEQ ID NO. 1, serine corresponding to position 31 of SEQ ID NO. 1, tyrosine corresponding to position 32 of SEQ ID NO. 1, methionine corresponding to position 34 of SEQ ID NO. 1, isoleucine corresponding to position 51 of SEQ ID NO. 1, asparagine corresponding to position 59 of SEQ ID NO. 1, alanine corresponding to position 97 of SEQ ID NO. 1, arginine corresponding to position 98 of SEQ ID NO. 1, glycine corresponding to position 99 of SEQ ID NO. 1, proline corresponding to position 101 of SEQ ID NO. 1, aspartate corresponding to position 102 of SEQ ID NO. 1, and SEQ ID NO. 1 Tyrosine corresponding to 103, lysine corresponding to position 108 of SEQ ID NO. 1, tyrosine corresponding to position 110 of SEQ ID NO. 1, phenylalanine corresponding to position 111 of SEQ ID NO. 1, aspartate corresponding to position 112 of SEQ ID NO. 1, arginine corresponding to position 40 of SEQ ID NO. 1, isoleucine corresponding to position 48 of SEQ ID NO. 1, lysine corresponding to position 67 of SEQ ID NO. 1, alanine corresponding to position 68 of SEQ ID NO. 1, leucine corresponding to position 70 of SEQ ID NO. 1, valine corresponding to position 72 of SEQ ID NO. 1, lysine corresponding to position 74 of SEQ ID NO. 1, alanine corresponding to position 79 of SEQ ID NO. 1, glutamine corresponding to position 82 of SEQ ID NO. 1, threonine corresponding to position 87 of SEQ ID NO. 1, serine corresponding to position 91 of SEQ ID NO. 1, or any combination thereof and / or SEQ ID NO. A mutation corresponding to position 33 of 1, e.g., W33H, a mutation corresponding to position 50 of sequence number 1,For example, E50H, a mutation corresponding to position 52 of SEQ ID NO. 1, for example, N52H, a mutation corresponding to position 53 of SEQ ID NO. 1, for example, P53H, a mutation corresponding to position 54 of SEQ ID NO. 1, for example, S54H, a mutation corresponding to position 55 of SEQ ID NO. 1, for example, N55H, a mutation corresponding to position 56 of SEQ ID NO. 1, for example, G56H, a mutation corresponding to position 57 of SEQ ID NO. 1, for example, R57H, a mutation corresponding to position 58 of SEQ ID NO. 1, for example, T58H, a mutation corresponding to position 60 of SEQ ID NO. 1, for example, Y60H, a mutation corresponding to position 61 of SEQ ID NO. 1, for example, N61H, a mutation corresponding to position 62 of SEQ ID NO. 1, for example, Q62H, a mutation corresponding to position 63 of SEQ ID NO. 1, for example, K63H, a mutation corresponding to position 64 of SEQ ID NO. 1, for example, F64H, a mutation corresponding to position 65 of SEQ ID NO. 1, e.g., Q65H, a mutation corresponding to position 66 of SEQ ID NO. 1, e.g., G66H, a mutation corresponding to position 96 of SEQ ID NO. 1, e.g., F96H, a mutation corresponding to position 100 of SEQ ID NO. 1, e.g., R100H, a mutation corresponding to position 104 of SEQ ID NO. 1, e.g., Y104H, a mutation corresponding to position 105 of SEQ ID NO. 1, e.g., G105H, a mutation corresponding to position 106 of SEQ ID NO. 1, e.g., S106H, a mutation corresponding to position 107 of SEQ ID NO. 1, e.g., S107H, a mutation corresponding to position 109 of SEQ ID NO. 1, e.g., W109H, a mutation corresponding to position 113 of SEQ ID NO. 1, e.g., V113H, a mutation corresponding to position 12 of SEQ ID NO. 1, e.g., V12K, SEQ ID NO. 1 position A mutation corresponding to 20, e.g., L20V; a mutation corresponding to position 22 of SEQ ID NO. 1, e.g., C22A or C22K; a mutation corresponding to position 38 of SEQ ID NO. 1, e.g., K38R,Includes a mutation corresponding to position 76 of sequence number 1, e.g., S76T or any combination thereof. In some embodiments, the variable light chain is isoleucine corresponding to position 29 of SEQ ID NO. 2, valine corresponding to position 34 of SEQ ID NO. 2, glutamate corresponding to position 39 of SEQ ID NO. 2, asparagine corresponding to position 58 of SEQ ID NO. 2, glycine corresponding to position 96 of SEQ ID NO. 2, proline corresponding to position 100 of SEQ ID NO. 2, proline corresponding to position 101 of SEQ ID NO. 2, threonine corresponding to position 102 of SEQ ID NO. 2, threonine corresponding to position 7 of SEQ ID NO. 2, serine corresponding to position 14 of SEQ ID NO. 2, aspartate corresponding to position 17 of SEQ ID NO. 2, leucine corresponding to position 42 of SEQ ID NO. 2, lysine corresponding to position 44 of SEQ ID NO. 2, leucine corresponding to position 51 of SEQ ID NO. 2, alanine corresponding to position 72 of SEQ ID NO. 2, and corresponding to position 79 of SEQ ID NO. 2. Arginine or any combination thereof and / or a mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, e.g., T36H; a mutation corresponding to position 38 of SEQ ID NO. 2, e.g., L38H; a mutation corresponding to position 55 of SEQ ID NO. 2, e.g., K55H, a mutation corresponding to position 56 of sequence number 2, e.g., V56H,Includes a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; a mutation corresponding to position 90 of SEQ ID NO. 2, e.g., I90V; or any combination thereof.

[0008] The present invention provides an antibody comprising a variable heavy chain having an amino acid sequence at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence at least 90% identical to SEQ ID NO. 2, wherein SEQ ID NO. 1 is a mutation at position 26, e.g., G26H; a mutation at position 27, e.g., Y27H; a mutation at position 28, e.g., T28H; a mutation at position 29, e.g., F29H; a mutation at position 30, e.g., T30H; a mutation at position 31, e.g., S31H; a mutation at position 32, e.g., Y32H; a mutation at position 34, e.g., M34H; a mutation at position 51, e.g., I51H; a mutation at position 59, e.g., N59H; a mutation at position 97, e.g., A97H; a mutation at position 98, e.g., R98H; a mutation at position 99, e.g., G99H; a mutation at position 101, e.g., P101H, mutation at position 102, e.g., D102H, mutation at position 103, e.g., Y103H, mutation at position 108, e.g., K108H, mutation at position 110, e.g., Y110H, mutation at position 111, e.g., F111H, mutation at position 112, e.g., D112H, mutation at position 40, e.g., R40A, mutation at position 48, e.g., I48M, mutation at position 67, e.g., K67R, mutation at position 68, e.g., A68V, mutation at position 70, e.g., L70M, mutation at position 72, e.g., V72R, mutation at position 74, e.g., K74T, mutation at position 79, e.g., A79V, mutation at position 82, e.g., Q82E, mutation at position 87, e.g., T87R, position excluding mutations at 91, e.g., S91T, SEQ ID NO. 2 includes mutations at position 29, e.g., I29H, mutations at position 34, e.g., V34H,It does not include mutations at position 37, e.g., Y37H; mutations at position 39, e.g., E39H; mutations at position 58, e.g., N58H; mutations at position 96, e.g., G96H; mutations at position 100, e.g., P100H; mutations at position 101, e.g., P101H; mutations at position 102, e.g., T102H; mutations at position 7, e.g., T7S; mutations at position 14, e.g., S14T; mutations at position 17, e.g., D17Q; mutations at position 42, e.g., L42Q; mutations at position 44, e.g., K44R; mutations at position 51, e.g., L51R; mutations at position 72, e.g., A72S; mutations at position 79, e.g., R79K. In some embodiments, the variable heavy chain is a mutation corresponding to position 33 of SEQ ID NO. 1, e.g., W33H; a mutation corresponding to position 50 of SEQ ID NO. 1, e.g., E50H; a mutation corresponding to position 52 of SEQ ID NO. 1, e.g., N52H; a mutation corresponding to position 53 of SEQ ID NO. 1, e.g., P53H; a mutation corresponding to position 54 of SEQ ID NO. 1, e.g., S54H; a mutation corresponding to position 55 of SEQ ID NO. 1, e.g., N55H; a mutation corresponding to position 56 of SEQ ID NO. 1, e.g., G56H; a mutation corresponding to position 57 of SEQ ID NO. 1, e.g., R57H; a mutation corresponding to position 58 of SEQ ID NO. 1, e.g., T58H; a mutation corresponding to position 60 of SEQ ID NO. 1, e.g., Y60H; a mutation corresponding to position 61 of SEQ ID NO. 1, e.g., N61H; a mutation corresponding to position 62 of SEQ ID NO. 1, e.g., Q62H, a mutation corresponding to position 63 of SEQ ID NO. 1, e.g., K63H, a mutation corresponding to position 64 of SEQ ID NO. 1, e.g., F64H, a mutation corresponding to position 65 of SEQ ID NO. 1, e.g., Q65H,A mutation corresponding to position 66 of SEQ ID NO. 1, e.g., G66H; a mutation corresponding to position 96 of SEQ ID NO. 1, e.g., F96H; a mutation corresponding to position 100 of SEQ ID NO. 1, e.g., R100H; a mutation corresponding to position 104 of SEQ ID NO. 1, e.g., Y104H; a mutation corresponding to position 105 of SEQ ID NO. 1, e.g., G105H; a mutation corresponding to position 106 of SEQ ID NO. 1, e.g., S106H; a mutation corresponding to position 107 of SEQ ID NO. 1, e.g., S107H; a mutation corresponding to position 109 of SEQ ID NO. 1, e.g., W109H; a mutation corresponding to position 113 of SEQ ID NO. 1, e.g., V113H; a mutation corresponding to position 12 of SEQ ID NO. 1, e.g., V12K; a mutation corresponding to position 20 of SEQ ID NO. 1, e.g., L20V; a mutation corresponding to position 22 of SEQ ID NO. 1 It includes mutations, e.g., C22A or C22K, mutations corresponding to position 38 of SEQ ID NO. 1, e.g., K38R, mutations corresponding to position 76 of SEQ ID NO. 1, e.g., S76T, or any combination thereof. In some embodiments, the variable light chain is a mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, e.g., T36H; a mutation corresponding to position 38 of SEQ ID NO. 2, e.g., L38H,A mutation corresponding to position 55 of SEQ ID NO. 2, e.g., K55H; a mutation corresponding to position 56 of SEQ ID NO. 2, e.g., V56H; a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; SEQ ID NO. 2, position Includes mutations corresponding to 90, e.g., I90V or any combination thereof.

[0009] In some embodiments, the variable heavy chain comprises a W33H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 10.

[0010] In some embodiments, the variable heavy chain comprises an E50H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 12.

[0011] In some embodiments, the variable heavy chain comprises an N52H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 14.

[0012] In some embodiments, the variable heavy chain comprises a P53H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 15.

[0013] In some embodiments, the variable heavy chain comprises an S54H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 16.

[0014] In some embodiments, the variable heavy chain comprises an N55H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 17.

[0015] In some embodiments, the variable heavy chain comprises a G56H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 18.

[0016] In some embodiments, the variable heavy chain comprises an R57H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 19.

[0017] In some embodiments, the variable heavy chain comprises a T58H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 20.

[0018] In some embodiments, the variable heavy chain comprises a Y60H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 22.

[0019] In some embodiments, the variable heavy chain comprises an N61H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 23.

[0020] In some embodiments, the variable heavy chain comprises a Q62H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 24.

[0021] In some embodiments, the variable heavy chain comprises a K63H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 25.

[0022] In some embodiments, the variable heavy chain comprises an F64H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 26.

[0023] In some embodiments, the variable heavy chain comprises a Q65H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 27.

[0024] In some embodiments, the variable heavy chain comprises a G66H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 28.

[0025] In some embodiments, the variable heavy chain comprises an F96H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 29.

[0026] In some embodiments, the variable heavy chain comprises an R100H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 33.

[0027] In some embodiments, the variable heavy chain comprises a Y104H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 37.

[0028] In some embodiments, the variable heavy chain comprises a G105H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 38.

[0029] In some embodiments, the variable heavy chain comprises an S106H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 39.

[0030] In some embodiments, the variable heavy chain comprises an S107H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 40.

[0031] In some embodiments, the variable heavy chain comprises a W109H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 42.

[0032] In some embodiments, the variable heavy chain comprises a V113H mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 46.

[0033] In some embodiments, the variable heavy chain comprises a V12K mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 77.

[0034] In some embodiments, the variable heavy chain comprises an L20V mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 78.

[0035] In some embodiments, the variable heavy chain comprises a C22A or C22K mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 79 or 80.

[0036] In some embodiments, the variable heavy chain comprises a K38R mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 81.

[0037] In some embodiments, the variable heavy chain comprises an S76T mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 89.

[0038] In some embodiments, the variable light chain comprises an R24H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 47.

[0039] In some embodiments, the variable light chain comprises an S25H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 48.

[0040] In some embodiments, the variable light chain comprises an S26H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 49.

[0041] In some embodiments, the variable light chain comprises a Q27H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 50.

[0042] In some embodiments, the variable light chain comprises an S28H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 51.

[0043] In some embodiments, the variable light chain comprises a V30H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 53.

[0044] In some embodiments, the variable light chain comprises an S32H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 54.

[0045] In some embodiments, the variable light chain comprises an N33H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 55.

[0046] In some embodiments, the variable light chain comprises an N35H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 57.

[0047] In some embodiments, the variable light chain comprises a T36H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 58.

[0048] In some embodiments, the variable light chain comprises an L38H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 60.

[0049] In some embodiments, the variable light chain comprises a K55H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 62.

[0050] In some embodiments, the variable light chain comprises a V56H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 63.

[0051] In some embodiments, the variable light chain comprises an S57H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 64.

[0052] In some embodiments, the variable light chain comprises an R59H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 66.

[0053] In some embodiments, the variable light chain comprises an F60H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 67.

[0054] In some embodiments, the variable light chain comprises an S61H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 68.

[0055] In some embodiments, the variable light chain comprises an F94H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 69.

[0056] In some embodiments, the variable light chain comprises a Q95H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 70.

[0057] In some embodiments, the variable light chain comprises an S97H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 72.

[0058] In some embodiments, the variable light chain comprises a V99H mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 73.

[0059] In some embodiments, the variable light chain comprises a Y41F mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 97.

[0060] In some embodiments, the variable light chain comprises an L88V mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 103.

[0061] In some embodiments, the variable light chain comprises an I90V mutation. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 104.

[0062] In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 10. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 12. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 14. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 15. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 16. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 17. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 18. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 19. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 20. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 22. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 23. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 24. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 25. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 26.In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 27. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 28. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 29. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 33. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 37. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 38. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 39. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 40. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 42. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 46. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 77. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 78. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 79. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 80.In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 81. In some embodiments, the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 89.

[0063] In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 47. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 48. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 49. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 50. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 51. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 53. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 54. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 55. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 57. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 58. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 60. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 62. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 63. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 64.In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 66. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 67. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 68. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 69. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 70. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 72. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 73. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 97. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 103. In some embodiments, the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 104.

[0064] The present invention provides a method for treating thyroid-associated eye disease (TAO) or its symptoms or reducing its severity, which includes administering the antibody provided in the present invention to a subject.

[0065] The present invention provides a method for reducing proptosis in the eyes of a subject having thyroid-associated eye disease (TAO), comprising administering the antibody provided in the present invention to the subject.

[0066] The present invention provides a method for treating thyroid eye disease in a subject, which includes administering an antibody provided in the present invention to the subject.

[0067] The present invention provides a method for reducing the Clinical Activity Score (CAS) of thyroid-associated eye disease (TAO) in a subject, which includes administering the antibody provided in the present invention to the subject.

[0068] The present invention provides a method for a) reducing proptosis by at least 2 mm and b) reducing the clinical activity score (CAS) in subjects with thyroid-associated eye disease (TAO), which includes administering the antibody provided in the present invention to the subjects.

[0069] In some embodiments, the protrusion of the eyeball is reduced by at least 2 mm. In some embodiments, the protrusion of the eyeball is reduced by at least 3 mm. In some embodiments, the protrusion of the eyeball is reduced by at least 4 mm. In some embodiments, 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) point. In some embodiments, the subject's clinical activity score (CAS) is reduced to zero (0) point.

[0070] The present invention provides a method for treating thyroid-associated eye disease (TAO) or reducing its severity in a subject, comprising administering the antibody provided in the present invention to the subject, wherein treatment using said antibody (i) reduces proptosis of one eyeball by at least 2 mm, (ii) does not entail worsening of the opposite eyeball (or opposite eye) by more than 2 mm, and (iii) reduces the subject's CAS to one (1) or zero (0).

[0071] The present invention provides a method for improving the quality of life in a subject having thyroid-associated eye disease (TAO, also known as Graves' ophthalmopathy / Graves' orbital disease), which comprises administering the antibody provided herein to the subject. In some embodiments, the quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment, or by either the visual function or appearance subscales thereof. In some embodiments, treatment results in an improvement of 8 points or more in the GO-QoL. In some embodiments, treatment results in an improvement in the function subscale of the GO-QoL. In some embodiments, treatment results in an improvement in the appearance subscale of the GO-QoL.

[0072] The present invention provides a method for treating diplopia or reducing its severity in subjects with thyroid-associated eye disease (TAO), comprising administering the antibody provided herein to the subject. In some embodiments, the diplopia is persistent diplopia. In some embodiments, the diplopia is non-persistent diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, improvement in diplopia or reduction in its severity is maintained for at least 20 weeks after discontinuation of antibody administration. In some embodiments, improvement in diplopia or reduction in its severity is maintained for at least 50 weeks after discontinuation of antibody administration.

[0073] 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 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 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 antibody. In some embodiments, the antibody is administered as a first dose at a dosage of about 10 mg / kg of antibody and as a subsequent dose at a dosage of about 20 mg / kg of antibody. In some embodiments, the subsequent dose is administered every 3 weeks for at least 21 weeks. In some embodiments, the antibody or its antigen-binding fragment is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a diabody, a single-chain antibody, a multi-specific antibody, Fab, Fab', F(ab')2, Fv, or scFv. In some embodiments, the antibody or its antigen-binding fragment is administered in a pharmaceutical composition comprising additionally a pharmaceutically acceptable diluent, excipient, or carrier. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. In some embodiments, 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. In some embodiments, the antibody or its antigen-binding fragment is administered directly into the eye, the anterior chamber of the eye, the vitreous cavity of the eye, the supracoracial space, or the retroorbital sinus. In some embodiments, the antibody or its antigen-binding fragment is administered by injection. In some embodiments, the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a supracoracial injection, or an anterior injection.

[0074] The present invention provides a method for increasing the internalization of IGF-1R in cells, the method comprising contacting the cells with the antibody provided herein. In some embodiments, contacting comprises administering the antibody provided herein to a subject. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof.

[0075] The present invention provides a method for inhibiting IGF-1-stimulated receptor phosphorylation on cells, the method comprising contacting the cells with the antibody provided herein. In some embodiments, contacting comprises administering the antibody provided herein to a subject. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof. In some embodiments, the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, or 0.09 nm or less. In some embodiments, the IC50 is measured in an in vitro assay, such as the assay provided herein. In some embodiments, the cells are A549 cells or HOCF cells.

[0076] The present invention provides a method for treating thyroid eye disease in a subject, the method comprising administering the antibody provided herein, wherein the antibody has a serum concentration in the subject of 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 after at least 1, 2, or 3 weeks after administration. In some embodiments, the antibody is administered intravenously. In some embodiments, the antibody is administered at a dose of about 20 mg / kg. In some embodiments, the antibody is administered at least or about once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks.

[0077] The present invention provides a method for inhibiting IGF-1-induced receptor autophosphorylation in cells by at least 95%, 96%, 97%, 98%, 99%, or 100%, the method comprising contacting the cells with the antibody provided herein. In some embodiments, inhibition of IGF-1-induced receptor autophosphorylation is measured in comparison to receptor autophosphorylation induced in the absence of the antibody. In some embodiments, contacting comprises administering the antibody comprising the above to a subject. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof.

[0078] The present invention provides a method for inhibiting IGF-1-induced receptor autophosphorylation in a subject in need by at least 95%, 96%, 97%, 98%, 99%, or 100%, and the method comprises administering the antibody provided in the present invention to the subject. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof. In some embodiments, the antibody is administered intravenously. Specific details for implementing the invention

[0079] The present invention provides an antibody that binds to IGF-1R and regulates its activity. The antibody can be used, for example, to treat thyroid eye disease.

[0080] As used herein, ‘thyroid-associated eye disease’ (TAO), ‘thyroid eye disease’ (TED), ‘Graves’ eye disease’, or ‘Graves’ orbital disease’ (GO) refer to the same disorder or condition and are used interchangeably. 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.

[0081] The terms 'exophthalmos' and 'exophthalmos' (also known as exophthalmos, exophthalmia, or exorbitism) refer to the forward projection, displacement, bulging, or protrusion of an organ. As used herein, the terms refer to forward projection, displacement, bulging, or protrusion anterior to the outside of the orbit. While exophthalmos and exophthalmos are considered to have the same meaning and are often used interchangeably by those skilled in the art in some fields, others assign subtle differences to their meanings. Exophthalmos is used by some to refer to severe exophthalmos or endocrine-related exophthalmos. Still others use the term exophthalmos when describing exophthalmos associated with the eye in subjects with TAO (TED or GO), for example.

[0082] As used herein, the terms 'propocular projection' and 'propocular protrusion' are used interchangeably and refer to anterior projection, displacement, protrusion, or bulge outward from the orbit. Due to the rigid bony structure of the orbit, which has only an anterior opening for expansion, any increase in orbital soft tissue contents originating from the side or posterior side will displace the eyeball anteriorly. Propocular projection or propocular protrusion may be the result of several disease processes, including infection, inflammation, tumors, trauma, metastasis, endocrine lesions, vascular diseases, and extraocular lesions. TAO (TED or GO) is currently recognized as the most common cause of propocular projection in adults. Propocular protrusion may be bilateral, as often observed in TAO (TED or GO), or unilateral (as commonly observed in orbital tumors).

[0083] The degree of exophthalmos can be measured, for example, using an exophthalmometer, an instrument used to measure the degree of anterior displacement of the eyeball. This device enables the measurement of the anterior distance of the lateral orbital margin toward the anterior surface of the cornea. Computed tomography (CT) scanning and magnetic resonance imaging (MRI) can also be used to evaluate the degree of exophthalmos or proptosis. CT scanning is an excellent imaging modality for the diagnosis of TAO. In addition to enabling the visualization of hypertrophied extraocular muscles, CT scans provide surgeons or clinicians with a description of the orbital bone structure when orbital decompression is required. Because MRI possesses multi-plane and intrinsic contrast capabilities, it provides excellent imaging of orbital contents without the radiation exposure associated with CT scan studies. While MRI provides better imaging of the optic nerve, orbital fat, and extraocular muscles, CT scans allow for better visualization of the orbital bone structure. Orbital ultrasound can also be used for the diagnosis and evaluation of TAO, as it can be performed rapidly and with a high degree of reliability. High reflectivity and hypertrophy of the extraocular muscles are easily assessed, and a series of ultrasound examinations may be used to evaluate the progression or stability of the ophthalmopathy. Based on currently available or future technologies, a person skilled in the art will be able to determine the best modality for diagnosing and evaluating the extent of proptosis.

[0084] As used herein, the term "antibody" refers to any form of antibody exhibiting the intended biological activity. Accordingly, it is used in its 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 (e.g., camelid antibodies). "Parent antibody" is an antibody obtained by exposing the immune system to an antigen before modifying the antibody for an intended use, such as the humanization of the antibody for use as a human therapeutic antibody.

[0085] 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, e.g., a fragment retaining one or more CDR regions. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments; diabadies; linear antibodies; single-chain antibody molecules, e.g., scFv; nanobodies; and multispecific antibodies formed from antibody fragments.

[0086] 'Fab fragment' is a C of one light chain and one heavy chain. H It consists of 1 and a variable region. The heavy chain of the Fab molecule cannot form a disulfide bond with another heavy chain molecule.

[0087] The 'Fc' region is the antibody's C H 1 and C H It contains two heavy chain fragments containing 2 domains. The two heavy chain fragments are connected by two or more disulfide bonds and C H 3 domains are held together by hydrophobic interactions.

[0088] In some embodiments, the antibody or antigen fragment of the present invention comprises an Fc region. In some embodiments, the Fc region comprises a mutation that extends the half-life of the antibody when ligated 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, 'YTE mutations') can be found in the sequence of SEQ ID NO. 89. In some embodiments, an Fc region comprising YTE mutations comprises the sequence of SEQ ID NO. 90, which differs from SEQ ID NO. 89 by the presence of a C-terminal lysine (K) residue. The numbering of the Fc area may follow the Kabat numbering system for the Fc area.

[0089] In some embodiments, the Fc region includes S228P and L235E mutations. In some embodiments, the antibody includes L234F, L235E, and P331S mutations. In some embodiments, the Fc region includes M252Y, S254T, T256E, S228P, and L235E mutations. In some embodiments, the Fc region includes S228P, L235E, M428L, and N434S mutations. In some embodiments, the Fc region includes M428L and N434S mutations. In some embodiments, the Fc region includes L234F, L235E, P331S, M252Y, S254T, and T256E mutations. Mutations within the Fc region are also US2007041972A1, EP2235059B1, U.S. Patent No. 8,394,925 and the literature [Mueller et al. (1997) Mol Immunol.[34(6):441-52] is described, each of which is included by reference in its entirety. The numbering referenced herein refers to the Kabat numbering system for the Fc area.

[0090] In some embodiments, the Fc region includes the following sequence.

[0091] APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 199),

[0092] APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 200),

[0093] APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 201),

[0094] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(서열번호 202),

[0095] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK(서열번호 203),

[0096] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(서열번호 204),

[0097] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(서열번호 205) 또는

[0098] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 206).

[0099] The 'Fab' short story is a single light chain and V H Domain and C H 1 domain and also C H 1 Domains and C H It contains a portion or fragment of a heavy chain containing a region between two domains, so that interchain disulfide bonds are formed between the two heavy chains of the two Fab' fragments to form an F(ab')2 molecule.

[0100] 'F(ab')2 Short Story' consists of 2 light chains and C H 1 Domains and C H 2 It contains two heavy chains containing a portion of the invariant region between the domains, so that an interchain disulfide bond is formed between the two heavy chains. Thus, the F(ab')2 fragment consists of two Fab' fragments held together by a disulfide bond between the two heavy chains.

[0101] The 'Fv region' includes variable regions from both heavy and light chains, but lacks invariant regions.

[0102] The terms 'single-chain Fv' or 'scFv' refer to antibodies V of antibodies H and V LIt refers to antibody fragments containing domains, wherein these domains exist within a single polypeptide chain. Generally, Fv polypeptides are V H and V L It additionally includes polypeptide linkers between domains, which allows scFv to form a desired structure for antigen binding. For a review of scFv, refer to the literature [Pluckthun (1994) The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315]. Also refer to International Patent Application Publication No. WO 88 / 01649 and U.S. Patents No. 4,946,778 and 5,260,203.

[0103] 'Domain antibodies' are immunologically functional immunoglobulin fragments containing only the variable region of the heavy chain or the variable region of the light chain. In some cases, two or more V H The region is covalently linked to a peptide linker to generate a bivalent domain antibody. The two Vs of the bivalent domain antibody H The region can target the same or different antigens.

[0104] 'Bivalent antibodies' contain two antigen-binding sites. In some cases, the two binding sites have the same antigen specificity. However, bivalent antibodies can be bispecific (see below).

[0105] In certain embodiments, the monoclonal antibodies of the present invention also include camellia single-domain antibodies. For example, the literature [Muyldermans et al. (2001) Trends Biochem. Sci . 26:230], [Reichmann et al. (1999) J. Immunol. Methods[231:25], International Publication No. WO 94 / 04678, International Publication No. WO 94 / 25591, and U.S. Patent No. 6,005,079. In some embodiments, the monoclonal antibody of the present invention has two Vs modified to form a single-domain antibody. H It is a single-domain antibody containing a domain.

[0106] As used herein, the term 'diabadi' refers to a small antibody fragment having two antigen-binding sites, and such fragment has a light chain variable domain (V L Heavy chain variable domain (V) connected to ) H Includes )(V H -V L or V L -V H ). By using a linker that is too short to allow pairing between two domains on the same chain, the domain is forced to pair with a complementary domain on another chain, creating two antigen-binding sites. Diabodies are, for example, EP 404,097, International Publication No. WO 93 / 11161 and literature [Holliger et al. (1993) Proc. Natl. Acad. Sci. USA It is described more fully in [90:6444-6448], each of which is incorporated herein by reference in its entirety. For the review of engineered antibody variants, generally [Holliger and Hudson (2005) Nat. Biotechnol Refer to . 23:1126-1136.

[0107] Typically, a variant antibody or antigen-binding fragment of the antibody provided herein retains at least 10% of its IGF-1R binding activity when activity is expressed on a molar basis (compared to the parent antibody being modified). In some embodiments, the variant antibody (or its antigen fragment) or the antigen-binding fragment of the antibody provided herein retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the IGF-1R binding affinity of the parent antibody. As described herein, the antibody or antigen-binding fragment of the present invention may include conserved or non-conserved amino acid substitutions that do not substantially alter its biological activity, and it is also intended that these may also be referred to as 'conserved variants' or 'function-conserving variants' of the antibody.

[0108] "Isolated antibody" refers to the purified state of a binding compound, and in this context, means that the molecule is substantially free of other biological molecules, such as nucleic acids, proteins, lipids, carbohydrates, or other substances such as cell debris and growth media. Generally, the term "isolated" is not intended to refer to the complete absence of such substances or the absence of water, buffers, or salts, unless they are present in amounts that substantially interfere with the experimental or therapeutic use of the binding compound described herein.

[0109] As used herein, the term 'monoclonal antibody' refers to a substantially homogeneous population of antibodies, that is, the antibody molecules constituting the population have identical amino acid sequences, except for possible naturally occurring mutations that may be present in small amounts. In contrast, conventional (polyclonal) antibody formulations typically contain multiple different antibodies having different amino acid sequences within their variable domains, particularly within their CDRs, which are often specific to different epitopes. The modifier 'monoclonal' indicates the characteristic of the antibody as being obtained from a substantially homogeneous population of antibodies and should not be interpreted as requiring the generation of the antibody by any particular method. For example, the monoclonal antibody used according to the present invention is [Kohler et al. (1975) Nature It can be produced by a hybridoma method first described in [256:495] or by a recombinant DNA method (see, e.g., U.S. Patent No. 4,816,567). 'Monoclonal antibodies' are also, for example, [Clackson et al. (1991) Nature 352:624-628], [Marks et al. (1991) J Mol Biol . 222:581-597] and [Presta LG (2005) J Allergy Clin Immunol It may also be isolated from a phage antibody library using the technique described in [116:731].

[0110] The 'chimeric antibody' used herein 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 U.S. Patent No. 4,816,567 and the literature [Morrison et al. (1984) Proc Natl Acad Sci. USAAs described in [81: 6851-6855], they are derived from different species. Typically, the variable domain is obtained from an antibody ('parent antibody') from an experimental animal such as a rodent, and the constant domain sequence is obtained from a human antibody, so the resulting chimeric antibody is less likely to induce an adverse immune response in human subjects than the parent (e.g., rodent) antibody.

[0111] As used herein, the term "humanized antibody" refers to a form of antibody containing sequences from both human and non-human (e.g., murine, rat) antibodies. Generally, a humanized antibody will contain substantially all of at least one, and typically two, variable domains, wherein all or substantially all of the hypervariable loop corresponds to that of a non-human immunoglobulin, and all or substantially all of the framework (FR) region is of a human immunoglobulin sequence. The humanized antibody may optionally contain at least a portion of the human immunoglobulin constant region (Fc).

[0112] The term 'fully human antibody' refers to an antibody containing only a human immunoglobulin protein sequence. Fully human antibodies may contain murine carbohydrate chains when produced in mice, mouse cells, or hybridomas derived from mouse cells. Similarly, 'mouse antibody' refers to an antibody containing only a mouse immunoglobulin sequence. Alternatively, fully human antibodies may contain rat carbohydrate chains when produced in rats, rat cells, or hybridomas derived from rat cells. Similarly, 'rat antibody' refers to an antibody containing only a rat immunoglobulin sequence.

[0113] Generally, the basic structural unit of an antibody comprises a tetramer. Each tetramer contains two identical polypeptide chain pairs, each pair having one 'light' chain (about 25 kDa) and one 'heavy' chain (about 50 to 70 kDa). The amino-terminal portion of each chain contains a variable region of about 100 to 110 or more amino acids, primarily responsible for antigen recognition. The carboxy-terminal portion of the heavy chain may define a constant region primarily responsible for effector functions. Typically, human light chains are classified into kappa and lambda light chains. Additionally, human heavy chains are typically classified into mu, delta, gamma, alpha, or epsilon, and the antibody isoforms are defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are united by a 'J' region of about 12 or more amino acids, and the heavy chain also contains a 'D' region of about 10 or more amino acids. Generally, the literature[ Fundamental Immunology See , Ch. 7 (Paul, W., ed., 2nd ed. Raven Press, NY (1989))].

[0114] The variable region of each light / heavy chain pair forms the antibody binding site. Therefore, generally, an intact antibody has two binding sites. Except for bifunctional or bispecific antibodies, the two binding sites are generally identical.

[0115] Typically, variable domains of both heavy and light chains contain three hypervariable regions, also known as complementarity determining regions (CDRs), located within a relatively conserved framework region (FR). The CDRs are generally aligned by the framework region to enable binding to specific epitopes. Generally, from the N-terminus to the C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The amino acid assignments for each domain are generally [North et al. , 'A new clustering of antibody CDR loop conformations,' J Mol Biol. It follows the definition in [ , 2011, Vol. 406, No. 2, pp. 228-256]. The assignment of amino acids to each domain is also [ Sequences of Proteins of Immunological Interest , Kabat et al.; National Institutes of Health, Bethesda, Md.; 5 th ed.; NIH Publ. 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. It may also follow the definition of [Chothia et al. (1989) Nature 342:878-883] or [Chothia et al. (1989) Nature 342:878-883].

[0116] As used herein, the term 'hypervariable region' refers to the amino acid residue of an antibody responsible for antigen-binding. The hypervariable region [North et al. , 'A new clustering of antibody CDR loop conformations,' J Mol Biol.It includes amino acid residues from the 'complementarity determining region' or 'CDR' defined in [Kabat, 2011, Vol. 406, No. 2, pp. 228-256] (i.e., residues between specific sequence motifs corresponding to the start and end positions of CDRL1, CDRL2, and CDRL3 within the light chain variable domain and the start and end positions of CDRH1, CDRH2, and CDRH3 within the heavy chain variable domain). The hypervariable region also includes [Kabat et al. Amino acid residues from the ‘complementarity determining region’ or ‘CDR’ [i.e., residues 24 to 34 (CDRL1), 50 to 56 (CDRL2) and 89 to 97 (CDRL3) in the light chain variable domain and residues 31 to 35 (CDRH1), 50 to 65 (CDRH2) and 95 to 102 (CDRH3) in the heavy chain variable domain] and / or literature [Chothia and Lesk (1987) J. Mol. Biol These residues from the 'hypervariable loop' defined in [North . 196: 901-917] may include residues 26 to 32 (CDRL1), 50 to 52 (CDRL2), and 91 to 96 (CDRL3) in the light chain variable domain and residues 26 to 32 (CDRH1), 53 to 55 (CDRH2), and 96 to 101 (CDRH3) in the heavy chain variable domain. Unless otherwise provided, any CDR provided herein [North et al.It is defined by the definition of CDR found in

[2011] . As used herein, the terms 'framework' or 'FR' residues refer to such variable domain residues other than the hypervariable domain residues defined herein as CDR residues. CDRs provide most of the contact residues for an antibody to bind to an antigen or epitope. CDRs of interest may be derived from donor antibody variable heavy chain and light chain sequences and include analogs of naturally occurring CDRs, which also share or retain the same antigen binding specificity and / or neutralizing ability as the donor antibody from which they are derived.

[0117] Additionally, in some embodiments, the antibody is a full-length antibody, a single-domain antibody, a recombinant heavy-chain-alone antibody (VHH), a single-chain antibody (scFv), a shark heavy-chain-alone antibody (VNAR), a microprotein (cysteine ​​knot protein, notin), DARPin; tetranectin; affibody; transbody; anticalin; adnectin; afilin; microbody; peptide aptamer; alterase; plastic antibody; filomer; stradobody; maxibody; avibody; fynomer, armadillo repeat protein, Kunitz domain, avimer, atrimer, probody, immunobody, triomab, troibody; pepbody; vaccibody, unibody; They may take the form of affimers, duobodies, Fv, Fab, Fab', F(ab')2, peptide mimic molecules, or synthetic molecules, which are U.S. Patent No. or Patent Publication No. US 7,417,130, US 2004 / 132094, US 5,831,012, US 2004 / 023334, US 7,250,297, US 6,818,418, US 2004 / 209243, US 7,838,629, US 7,186,524, US 6,004,746, US 5,475,096, US 2004 / 146938, US 2004 / 157209, US 6,994,982, US 6,794,144, US As described in 2010 / 239633, US 7,803,907, US 2010 / 119446 and / or US 7,166,697, the full text of each of which is incorporated herein by reference. Also reference [Storz MAbs. 2011 May-Jun; 3(3): 310-317], which is incorporated herein by reference.

[0118] As used herein, the term “antigen” means any molecule that has the ability to generate antibodies directly or indirectly or binds to antibodies. The definition of “antigen” includes protein-coding nucleic acids. “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 lysed or ruptured by the use of a detergent or other reagent. A cell treated with a detergent or other reagent that ruptures the cell membrane or punctures the cell membrane is not an intact cell. For example, a method for generating an antibody that binds to an IGF-1R protein is provided herein, the method comprising culturing a cell containing a nucleic acid molecule encoding an IGF-1R antibody.

[0119] As used herein, “specific binding,” “immunospecific binding,” or “binds immunospecifically” refers to the binding of an antibody to a predetermined antigen (e.g., IGF-1R) or an epitope present on the antigen. In some embodiments, the antibody is 10 -7 Dissociation constant less than or equal to M (K D It binds to ), and the K for binding to non-specific antigens other than predetermined antigens (e.g., BSA, casein, or another non-specific polypeptide). D K that is at least 2 times less than DIt binds to a predetermined antigen. The phrases 'antibody recognizing IGF-1R' and 'antibody specific to IGF-1R' are used interchangeably herein with the term 'antibody immunospecifically binding to IGF-1R'. IGF-1R may be referenced in this disclosure. The degree of specificity required for an anti-IGF-1R antibody may vary depending on the intended use of the antibody and is, in any case, defined by its suitability for use for the intended purpose. The antibody of the method considered in some embodiments, or a binding compound derived from the antigen-binding site of the antibody, binds to the antigen (IGF-1R) with an affinity that is at least 2 times greater, at least 10 times greater, at least 20 times greater, or at least 100 times greater than the affinity for any other antigen.

[0120] Methods for determining mAb specificity and affinity by competitive inhibition are described in the literature [Harlow et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1988], [Colligan et al., eds., Current Protocols in Immunology, Greene Publishing Assoc. and Wiley Interscience, NY, (1992, 1993)] and [Muller (1983) Meth Enzymol. They can be found in [92:589-601], the full text of each of which is incorporated herein by reference.

[0121] The term 'homologous' means a protein sequence having 40% to 100% sequence homology or identity with respect to a reference sequence. Percent identity between two peptide chains may be determined by pairwise alignment using the default settings of the AlignX module of Vector NTI v.9.0.0 (Invitrogen Corp., Carlsbad, Calif.). In some embodiments, the antibody or its antigenic binding fragment has at least 50, 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% homology or identity with respect to the sequence described herein. In some embodiments, the antibody has a conservative substitution with respect to the sequence described herein. An exemplary conservative substitution is 표 1 Examples are provided and are covered within the scope of the disclosed subject matter. Conservative substitutions may be present in the framework region or antigen-binding site as long as they do not adversely affect the properties of the antibody. Substitutions may be made to improve antibody properties, e.g., stability or affinity. Conservative substitutions will produce molecules having functional and chemical characteristics similar to the molecule to which these modifications were made. Exemplary amino acid substitutions are as follows: 표 1 It appears in.

[0122]

[0123] In some embodiments, variants of the protein and peptide provided herein are provided. In some embodiments, the variant comprises substitutions, deletions, or insertions. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) substitutions. The substitutions described herein may be conservative substitutions. In some embodiments, the substitutions are non-conservative. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) deletions. In some embodiments, the variant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (e.g., 1 to 10) insertions. In some embodiments, the substitutions, deletions, or insertions are present in the CDR provided herein. In some embodiments, substitutions, deletions, or insertions are not present in the CDR provided herein.

[0124] As used herein, the term "in combination with" means that the described formulations may be administered to an animal or subject together as a mixture, simultaneously as single formulations, or sequentially as single formulations in any order.

[0125] Techniques for generating antibodies against small peptide sequences that recognize and bind to such sequences when presented as unique sequences in the context of large proteins, either in a free or conjugated form or in a large protein are known in the art. Such antibodies include murine, murine-human, and human-human antibodies generated by hybridoma or recombinant techniques known in the art. Antibodies may also be generated in humans, mice, sheep, rats, rabbits, sharks, llamas, or chickens. In some embodiments, antibodies are generated in chickens. Antibodies may also be generated in or other small animals.

[0126] The term “epitope” is intended to refer to the corresponding portion of any molecule that can be recognized and bound by an antibody at one or more of the antigen-binding regions of the antibody. An epitope typically consists of a chemically active surface grouping of a molecule, such as an amino acid or a sugar side chain, and possesses specific three-dimensional structural features as well as specific charge features. Examples of epitopes include, but are not limited to, the residues described herein that form the IGF-1R epitope. In some embodiments, the epitope is present only in the non-denatured protein. In some embodiments, the epitope is present only in the denatured protein.

[0127] In some embodiments, the source for the DNA encoding the non-human antibody includes a cell line that produces the antibody, such as a hybrid cell line commonly known as a hybridoma.

[0128] Hybrid cells are formed by the fusion of animal splenocytes immunized with non-human antibody-generating cells, typically natural or recombinant antigens, or peptide fragments of antigen protein sequences. Alternatively, the non-human antibody-generating cells may be B lymphocytes obtained from the blood, spleen, lymph nodes, or other tissues of an animal immunized with an antigen.

[0129] The second fusion partner, which provides the immortalization function, may be a lymphoblastic cell, a plasmacytoma cell, or a myeloma cell, which is not an antibody-producing cell in itself but is malignant. The fusion partner cell includes, but is not limited to, hybridoma SP2 / 0-Ag14, abbreviated as SP2 / 0 (ATCC CRL1581), and myeloma P3X63Ag8 (ATCC TIB9) or derivatives thereof. For example, see Ausubel, Harlow, and Colligan below, the contents of which are incorporated herein by reference in their entirety.

[0130] Antibodies may be generated according to the embodiments provided herein. If the sequence is known, antibodies may also be generated according to known methods. Antibodies may also be converted into different types, such as human IgG, etc. By converting antibodies into human antibodies, human subjects should not identify the antibodies as foreign substances. The conversion of non-human IgG antibodies into human IgG antibodies is known and can be performed routinely once the specific sequence is known. The antibodies discussed herein may be modified according to known methods. Such methods are, for example, described in the literature [Riechmann et al. (1988) 'Reshaping human antibodies for therapy,' Nature 332(6162):332-323], [Tsurushita et al. (2004) ‘Humanization of a chicken anti-IL-12 monoclonal antibody,’ J Immunol Method. This is described in [295(1-2):9-19]. Antibody-producing cells that provide the nucleotide sequence encoding the antigen-binding domain of a chimeric antibody can also be generated by the transformation of non-human, such as primates, or human cells. For example, antibody-producing B lymphocytes can be infected with a virus such as the Epstein-Barr virus and transformed to yield immortal antibody-producing cells (Reference [Kozbor et al. (1983) Immunol. Today[4:72-79]). Alternatively, B lymphocytes may be transformed by providing a transforming gene or a transforming gene product, as is known in the art. For example, see Ausubel, Harlow, and Colligan below, the contents of which are incorporated herein by reference in their entirety. Cell fusion is achieved by standard procedures known to those skilled in the art of immunology. Fusion partner cell lines and methods for fusing and selecting hybridomas and screening mAbs are known in the art. For example, see Ausubel, Harlow, and Colligan below, the contents of which are incorporated herein by reference in their entirety.

[0131] In some embodiments, the antibody is an mAb that binds to IGF-1R. In some embodiments, the antibody binds to an amino acid of an epitope of IGF-1R.

[0132] In some embodiments, the antibody comprises the sequence provided herein.

[0133] The antibody sequence can be modified to yield human IgG antibodies. Transformations of the sequence provided herein can be modified to yield other types of antibodies. The CDR can also be linked to other antibodies, proteins, or molecules to produce antibody fragments that bind to IGF-1R. These may take the form of antibody-drug conjugates ('ADCs'), multi-specific molecules, or chimeric antigen receptors. The CDR and antibody sequences provided herein are also humanized or fully humanized according to known methods. The sequences can also be prepared into the chimeric antibodies described herein.

[0134] In some embodiments, the antibody comprises an amino acid sequence comprising the sequence or fragment thereof provided herein. In some embodiments, the antibody comprises one or more amino acid sequences provided herein, the antigen-binding fragment thereof, or a human IgG variant thereof. “Its human IgG variant” refers to an antibody modified to become human IgG when the starting antibody is not a human IgG antibody.

[0135] As described herein, the generation of antibodies having known sequences is routine and can be accomplished by any method. Accordingly, in some embodiments, a nucleic acid encoding an antibody or a fragment thereof is provided. In some embodiments, the nucleic acid encodes the sequence provided herein. The antibody may also be modified to become a chimeric antibody or a human antibody. The antibody may also be used in an injectable pharmaceutical composition. Additionally, the antibody described herein may be an isolated antibody or a engineered antibody.

[0136] In some embodiments, 'derivatives' of antibodies, fragments, regions, or derivatives thereof are provided, the terms include proteins encoded by genes that have been cut or modified to yield molecular species 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.

[0137] The identification of these antigen-binding regions and / or epitopes recognized by the antibodies described herein provides information necessary to generate additional monoclonal antibodies having similar binding characteristics and therapeutic or diagnostic utility corresponding to the embodiments of the present application.

[0138] The nucleic acid sequence encoding the antibody described herein may be genomic DNA or cDNA, or RNA (e.g., mRNA) encoding at least one of the variable regions described herein. A convenient alternative to using chromosomal gene fragments as a source of DNA encoding the V region antigen-binding segment is to use cDNA for the construction of chimeric immunoglobulin genes, which, for example, [Liu et al. (1987) Proc. Natl. Acad. Sci., USA 84:3439] and [Liu et al. (1987) J. Immunology As reported by [139:3521], the full text of which is incorporated herein by reference. The use of cDNA requires that a gene expression element appropriate to the host cell be combined with the gene to achieve the synthesis of the desired protein. The use of cDNA sequences is advantageous over genome sequences (which contain introns) because cDNA sequences can be expressed in bacteria or other hosts lacking an appropriate RNA splicing system.

[0139] For example, cDNA encoding a V region antigen-binding segment capable of detecting, binding to, or neutralizing the IGF-1R antigen may be provided using known methods based on the use of the amino acid sequence provided herein. Since the genetic code is degenerate, more than one codon may be used to encode a specific amino acid (Watson et al. below). Using the genetic code, one or more different oligonucleotides may be identified, each of which may encode an amino acid. The probability that a specific oligonucleotide actually constitutes the actual XXX-coding sequence can be estimated by considering anomalous base pairing relationships and the frequency with which a specific codon is actually used (to encode a specific amino acid) in eukaryotic or prokaryotic cells expressing antibodies or fragments. These 'codon usage rules' are [Lathe et al. (1985) J Molec Biol. [183:1-12] is disclosed. Using Lathe’s ‘codon usage rules,’ a single oligonucleotide or a set of oligonucleotides containing the theoretical ‘most probable’ nucleotide sequence capable of encoding the antibody variable or constant region sequence is identified.

[0140] The variable regions described herein may be combined with any type of constant region, including human constant regions or murine constant regions. The human gene encoding the constant (C) region of the antibody, fragment, and region may be derived from a human fetal liver library by known methods. The human C region gene may be derived from any human cell, including those that express and produce human immunoglobulin. Human C H The region may be derived from any of the known classes or isoforms of the human H chain, including gamma, μ, α, δ, or ε and their subtypes, e.g., G1, G2, G3, and G4. Since H chain isoforms are responsible for various effector functions of the antibody, C H The selection of the region will be guided by the desired effector function, such as activity in complement fixation or antibody-dependent cellular cytotoxicity (ADCC). Preferably, C H The region is derived from gamma 1 (IgG1), gamma 3 (IgG3), gamma 4 (IgG4), or μ (IgM). Human C LThe region may be derived from either kappa or lambda, which are human L-chain isoforms. In some embodiments, the antibody comprises an Fc domain. In some embodiments, the Fc domain comprises a mutation that extends 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, the full text of which is incorporated herein by reference. In some embodiments, the constant domain comprises a mutation at amino acid residue position 428 of the wild-type human IgG constant domain, numbered according to the EU numbering index of Kabat. Without intending to be bound by any theory, an antibody comprising the mutation corresponding to residue 428 may have an increased half-life compared to the half-life of IgG having the wild-type human IgG constant domain. In some embodiments, the mutation is a substitution of the intrinsic residue with threonine, leucine, phenylalanine, or serine. In some embodiments, the antibody further comprises one or more amino acid substitutions for the corresponding wild-type human IgG constant domain at one or more of amino acid residues 251 to 256, 285 to 290, 308 to 314, 385 to 389, and 429 to 436, 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, the full text of which is incorporated herein by reference.

[0141] Genes encoding the human immunoglobulin C region can be obtained from human cells by standard cloning techniques (see literature [Sambrook, et al. (Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (1989)] and [Ausubel et al., eds. Current Protocols in Molecular Biology (1987, 1993)]). Human C region genes are readily available from known clones containing genes representing two classes of the L chain, five classes of the H chain, and their subclasses. Chimeric antibody fragments such as F(ab')2 and Fab can be prepared by designing appropriately cleaved chimeric H chain genes. For example, a chimeric gene encoding the H chain portion of the F(ab')2 fragment will contain a DNA sequence encoding the CH1 domain and hinge region of the H chain, followed by a translation stop codon to yield the cleaved molecule.

[0142] In some embodiments, the antibodies, murine, human, humanized, or chimeric antibodies, fragments, and regions of the antibodies described herein clone DNA segments encoding the H and L chain antigen-binding regions of an IGF-1R antigen-specific antibody, and these DNA segments C H and C L It is generated by creating murine, human, or chimeric immunoglobulin-coding genes by combining DNA segments that each encode a region.

[0143] Accordingly, in some embodiments, a fused chimeric gene is produced comprising a first DNA segment encoding at least an antigen-binding region of non-human origin, such as a functionally rearranged V region having a union (J) segment connected to a second DNA segment encoding at least a portion of a human C region.

[0144] Accordingly, a method for generating an antibody according to some of the embodiments described herein, cDNA encoding antibody V and C regions, comprises several steps illustrated below: 1. isolation of messenger RNA (mRNA) from a cell line generating an anti-IGF-1R antigen antibody and from any additional antibody supplying heavy and light chain constant regions; cloning and cDNA generation therefrom; 2. preparation of a full-length cDNA library from the purified mRNA, whereby appropriate V and / or C region gene segments of L and H chain genes are (i) identified with appropriate probes, (ii) sequenced, and (iii) made compatible with C or V gene segments from another antibody for a chimeric antibody; 3. construction of a complete H or L chain coding sequence by linking the specific V region gene segment cloned as described above to the cloned C region gene; and 4. to provide murine-murine, human-murine, human-human, or human murine antibodies, it involves the expression and production of L and H chains in selected hosts, including prokaryotic and eukaryotic cells.

[0145] Two coding DNA sequences are referred to as 'operably linked' if the linkage results in a sequence that can be translated continuously without alteration or interruption of the triplet read frame. DNA coding sequences are operably linked to a gene expression element if the linkage causes the proper function of the gene expression element, thereby causing the expression of the coding sequence.

[0146] 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.

[0147] In some embodiments, the antibody described herein is used to detect the presence of an antigen. The antibody may be used in any device or method for detecting the presence of an antigen.

[0148] With respect to antibodies, the term 'purified' refers to an antibody that is substantially free of other substances that associate with molecules in its natural environment. For example, a purified protein is substantially free of cellular material or other proteins from the cell or tissue from which it originated. This term refers to a preparation in which the isolated protein is sufficiently pure for analysis, or is at least 70% to 80% (w / w) pure, at least 80% to 90% (w / w) pure, 90% to 95% pure, and at least 95%, 96%, 97%, 98%, 99%, or 100% (w / w) pure. In some embodiments, the antibody is purified.

[0149] As an alternative to manufacturing monoclonal antibody-secreting hybridomas, monoclonal antibodies against polypeptides can be identified and isolated by screening a recombinant immunoglobulin library (e.g., antibody-phage display library) against the polypeptide described herein, thereby isolating the immunoglobulin library members that bind to the polypeptide. Techniques and commercially available kits for generating and screening phage display libraries are known to those skilled in the art. Additionally, 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 may be used to screen other antibodies that can be used therapeutically, diagnostically, or as research tools.

[0150] 항체 접합체

[0151] 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 within the subject's body. Suitable polymers include, but are not limited to, polyethylene glycol (PEG) (e.g., PEG having a molecular weight of 2 kDa, 5 kDa, 10 kDa, 12 kDa, 20 kDa, 30 kDa, or 40 kDa), dextran, and monomethoxypolyethylene glycol (mPEG). Examples of PEG-conjugated single-chain antibodies are [Lee et al. (1999) Bioconj Chem. It can be found in [10:973-981], the full text of which is incorporated herein by reference. An example of conjugating antibodies with PEG attached to a radioactive metal chelating agent [diethylenetriaminepentaacetic acid (DTPA)] is found in the literature [Wen et al . (2001) Bioconj Chem.[12:545-553], the full text thereof is incorporated herein by reference. Examples of chemical moiety include, but are not limited to, anti-mitotic agents such as calichiamycin (e.g., ozogamicin), monomethylauristatin E, methansine, etc. Additional examples include, but are not limited to, biologically active anti-microtubule agents, alkylating agents, and DNA minor groove binders. Additional examples are provided herein and below. Chemical moiety may be linked to an antibody via a linker (e.g., maleimide), a cathepsin-cleavable linker (e.g., valine-citrulline), and in some embodiments, one or more spacers (e.g., para-aminobenzylcarbamate). Without intending to be bound by any particular theory, when an antibody conjugate binds to IGF-1R, it may be internalized, and the chemical moiety may kill the cell or otherwise inhibit its growth. In some embodiments, the cell is a thyroid cell.

[0152] The antibody and antibody fragment of the present invention also 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 It can also be combined with labels such as Fe.

[0153] Antibodies and antibody fragments are also rare earth chelates, fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanates, phycoerythrin, phycocyanin, allophycocyanin, o-phthalaldehyde, fluoressamine, 152 It may also be conjugated with fluorescent or chemiluminescent labels including fluorescent groups such as Eu, Dansil, Umbelliferone, Luciferin, Luminal label, Isoluminal label, Aromatic Acridinium ester label, Imidazole label, Acridinium salt label, Oxalate ester label, Equorin label, 2,3-Dihydrophthalazindione, Biotin / Avidin, Spin label, and stable free radical.

[0154] Antibody molecules may also be conjugated to cytotoxic factors such as diphtheria toxin, Pseudomonas aeruginosa exotoxin A chain, lysine A chain, abrin A chain, modesin A chain, alpha-sarcin, alleurates pordi proteins and compounds (e.g., fatty acids), diantin proteins, Phytolaca americana proteins PAPI, PAPII and PAP-S, Momordica charantia inhibitors, cursin, crotin, Saponaria officinalis inhibitors, mitogellin, restrictocin, phenomycin and enomycin.

[0155] Any method known in the art may be used to conjugate the antibody molecule of the present invention to various moiety, and this is [Hunter et al . (1962) Nature 144:945], [David et al ., (1974) Biochemistry 13:1014] and [Pain et al ., (1981) J Immunol Meth. 40:219] and [Nygren J (1982) Histochem and Cytochem.This includes the method described in [30:407]. Methods for conjugating antibodies are conventional and well known in the art.

[0156] 키메라 항원 수용체

[0157] The antibody provided herein may also be incorporated into a chimeric antigen receptor ('CAR') that can be used, for example, in CAR-T cells. In some embodiments, the extracellular domain of the CAR may be the antibody provided herein. In some embodiments, the antibody is in scFv format. CAR-T cells are a type of therapy in which a patient's T cells are modified to attack cells expressing IGF-1R. T cells are collected from the patient's blood. Then, a gene for a special receptor that binds to a specific protein on the patient's cells is added in the laboratory. In some embodiments, the receptor binds to IGF-1R using the binding domain of the antibody provided herein. Subsequently, CAR-T cells containing the IGF-1R antibody may be used to treat a pathological condition as provided herein.

[0158] In some embodiments, an antibody (e.g., an anti-IGF-1R antibody) is 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 natural (non-denatured) form. In some embodiments, the antibody does not specifically bind to a denatured IGF-1R protein. As used herein, the term "recombinant antibody" refers to an antibody that does not occur naturally. In some embodiments, the term "recombinant antibody" refers to an antibody that has not been isolated from a human subject.

[0159] In some embodiments, the antibody is as follows 표 2It may include a heavy chain variable region (VH) or light chain variable region (VL) sequence provided in, or a combination of VH and VL sequences, or variants.

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177] The heavy chains provided herein may be linked to an Fc region containing mutations that may affect the half-life of the antibody. Non-restrictive mutations within the Fc region are provided herein.

[0178] In some embodiments, the light chain region (LC) and heavy chain region (HC) may be combined with the VH and VL domains provided herein, which may or may not include an invariant region. The invariant region may be replaced as provided herein. The VH and VL regions may be used to form the antibodies provided herein. The VH and VL sequences may be any format, including but not limited to scFv formats in which the VH and VL regions are linked by peptide linkers. Examples of peptide linkers that may be used to link the various peptides provided herein are (GGGGS). n (Sequence No. 105), (GGGGA) n (SEQ ID NO. 106) or any combination thereof, but not limited to, wherein each n is independently 1 to 5. In some embodiments, the variable region is not connected by a peptide linker.

[0179] In some embodiments, the antibody or its antigen-binding fragment is SEQ ID NOs 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, It includes a heavy chain variable region (VH) sequence selected from the group consisting of 92 and 93.

[0180] In some embodiments, the antibody or its antigen-binding fragment comprises a light chain variable region (VL) sequence selected from the group consisting of SEQ ID NOs 2, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, and 104.

[0181] In some embodiments, an antibody is provided comprising a variable heavy chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 1 and a variable light chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 2, provided, provided,

[0182] Variable heavy chain

[0183] Glycine corresponding to position 26 of sequence number 1,

[0184] Tyrosine corresponding to position 27 of sequence number 1,

[0185] Threonine corresponding to position 28 of SEQ ID NO. 1,

[0186] Phenylalanine corresponding to position 29 of SEQ ID NO. 1,

[0187] Threonine corresponding to position 30 of SEQ ID NO. 1,

[0188] Serine corresponding to position 31 of sequence number 1,

[0189] Tyrosine corresponding to position 32 of sequence number 1,

[0190] Methionine corresponding to position 34 of SEQ ID NO. 1,

[0191] Isoleucine corresponding to position 51 of sequence number 1,

[0192] Asparagine corresponding to position 59 of sequence number 1,

[0193] Alanine corresponding to position 97 of sequence number 1,

[0194] Arginine corresponding to position 98 of sequence number 1,

[0195] Glycine corresponding to position 99 of sequence number 1,

[0196] Proline corresponding to position 101 of sequence number 1

[0197] Aspartate corresponding to position 102 of sequence number 1,

[0198] Tyrosine corresponding to position 103 of sequence number 1,

[0199] Lee Sin corresponding to position 108 of sequence number 1,

[0200] Tyrosine corresponding to position 110 of sequence number 1,

[0201] Phenylalanine corresponding to position 111 of SEQ ID NO. 1,

[0202] Aspartate corresponding to position 112 of sequence number 1,

[0203] Arginine corresponding to position 40 of SEQ ID NO. 1,

[0204] Isoleucine corresponding to position 48 of sequence number 1,

[0205] Lee Sin corresponding to position 67 of sequence number 1,

[0206] Alanine corresponding to position 68 of sequence number 1,

[0207] Leucine corresponding to position 70 of sequence number 1,

[0208] Valine corresponding to position 72 of sequence number 1,

[0209] Lee Sin corresponding to position 74 of sequence number 1,

[0210] Alanine corresponding to position 79 of sequence number 1,

[0211] Glutamine corresponding to position 82 of sequence number 1,

[0212] Threonine corresponding to position 87 of SEQ ID NO. 1,

[0213] Serine corresponding to position 91 of sequence number 1,

[0214] or includes any combination thereof.

[0215] In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises glycine corresponding to position 26 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises a tyrosine corresponding to position 27 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises threonine corresponding to position 28 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises phenylalanine corresponding to position 29 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises threonine corresponding to position 30 of SEQ ID NO. 1. In some embodiments, an antibody comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2 is provided, provided that the variable heavy chain comprises a serine corresponding to position 31 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises a tyrosine corresponding to position 32 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises methionine corresponding to position 34 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises isoleucine corresponding to position 51 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises asparagine corresponding to position 59 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises alanine corresponding to position 97 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises arginine corresponding to position 98 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises glycine corresponding to position 99 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises proline corresponding to position 101 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises aspartate corresponding to position 102 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises a tyrosine corresponding to position 103 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises lysine corresponding to position 108 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises a tyrosine corresponding to position 110 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises phenylalanine corresponding to position 111 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises aspartate corresponding to position 112 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises arginine corresponding to position 40 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises isoleucine corresponding to position 48 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises lysine corresponding to position 67 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises alanine corresponding to position 68 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises leucine corresponding to position 70 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises valine corresponding to position 72 of SEQ ID NO. 1 and lysine corresponding to position 74 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises alanine corresponding to position 79 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises glutamine corresponding to position 82 of SEQ ID NO. 1.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises threonine corresponding to position 87 of SEQ ID NO. 1. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable heavy chain comprises a serine corresponding to position 91 of SEQ ID NO. 1.

[0216] In some embodiments, an antibody is provided comprising a variable heavy chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 1 and a variable light chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 2, wherein the variable heavy chain is a mutation corresponding to position 33 of SEQ ID NO. 1 that is the same as but not limited to W33H, a mutation corresponding to position 50 of SEQ ID NO. 1 that is the same as but not limited to E50H, a mutation corresponding to position 52 of SEQ ID NO. 1 that is the same as but not limited to N52H, a mutation corresponding to position 53 of SEQ ID NO. 1 that is the same as but not limited to P53H, and a mutation corresponding to position 1 of SEQ ID NO. 1 that is the same as but not limited to S54H A mutation corresponding to 54, a mutation corresponding to position 55 of SEQ ID NO. 1 that is the same as but not limited to N55H, a mutation corresponding to position 56 of SEQ ID NO. 1 that is the same as but not limited to G56H, a mutation corresponding to position 57 of SEQ ID NO. 1 that is the same as but not limited to R57H, a mutation corresponding to position 58 of SEQ ID NO. 1 that is the same as but not limited to T58H, a mutation corresponding to position 60 of SEQ ID NO. 1 that is the same as but not limited to Y60H, a mutation corresponding to position 61 of SEQ ID NO. 1 that is the same as but not limited to N61H, a mutation corresponding to position 62 of SEQ ID NO. 1 that is the same as but not limited to Q62H, a mutation corresponding to position 63 of SEQ ID NO. 1 that is the same as but not limited to K63H, a mutation corresponding to position 64 of SEQ ID NO. 1 that is the same as but not limited to F64H, and a mutation corresponding to position 65 of SEQ ID NO. 1 that is the same as but not limited to Q65H. A mutation, identical but not limited to G66H, a mutation corresponding to position 66 of SEQ ID NO. 1, identical but not limited to F96H,A mutation corresponding to position 96 of SEQ ID NO. 1, identical to but not limited to R100H, a mutation corresponding to position 100 of SEQ ID NO. 1, identical to but not limited to Y104H, a mutation corresponding to position 104 of SEQ ID NO. 1, identical to but not limited to G105H, a mutation corresponding to position 105 of SEQ ID NO. 1, identical to but not limited to S106H, a mutation corresponding to position 106 of SEQ ID NO. 1, identical to but not limited to S107H, a mutation corresponding to position 107 of SEQ ID NO. 1, identical to but not limited to W109H, a mutation corresponding to position 109 of SEQ ID NO. 1, identical to but not limited to V113H, a mutation corresponding to position 113 of SEQ ID NO. 1, identical to but not limited to V12K, a mutation corresponding to position 12 of SEQ ID NO. 1, identical to but not limited to L20V, a mutation corresponding to position 20 of SEQ ID NO. 1 It includes a mutation corresponding to position 22 of SEQ ID NO. 1 that is equal to but not limited to C22A or C22K, a mutation corresponding to position 38 of SEQ ID NO. 1 that is equal to but not limited to K38R, a mutation corresponding to position 76 of SEQ ID NO. 1 that is equal to but not limited to S76T, or any combination thereof. In some embodiments, the variable heavy chain includes a mutation corresponding to position 33 of SEQ ID NO. 1 that is equal to but not limited to W33H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 50 of SEQ ID NO. 1 that is equal to but not limited to E50H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 52 of SEQ ID NO. 1 that is equal to but not limited to N52H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 53 of SEQ ID NO. 1 that is equal to but not limited to P53H. In some embodiments, the variable heavy chain includes, which is equal to but not limited to S54H,It includes a mutation corresponding to position 54 of SEQ ID NO. 1. In some embodiments, the variable heavy chain includes a mutation corresponding to position 55 of SEQ ID NO. 1, which is the same as but not limited to N55H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 56 of SEQ ID NO. 1, which is the same as but not limited to G56H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 57 of SEQ ID NO. 1, which is the same as but not limited to R57H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 58 of SEQ ID NO. 1, which is the same as but not limited to T58H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 60 of SEQ ID NO. 1, which is the same as but not limited to Y60H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 61 of SEQ ID NO. 1, which is the same as but not limited to N61H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 62 of SEQ ID NO. 1, which is the same as but not limited to Q62H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 63 of SEQ ID NO. 1, which is the same as but not limited to K63H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 64 of SEQ ID NO. 1, which is the same as but not limited to F64H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 65 of SEQ ID NO. 1, which is the same as but not limited to Q65H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 66 of SEQ ID NO. 1, which is the same as but not limited to G66H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 96 of SEQ ID NO. 1, which is the same as but not limited to F96H. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 100 of SEQ ID NO. 1, which is the same as but not limited to R100H. In some embodiments, the variable heavy chain is the same as Y104H but is not limited thereto,It includes a mutation corresponding to position 104 of SEQ ID NO. 1. In some embodiments, the variable heavy chain includes a mutation corresponding to position 105 of SEQ ID NO. 1, which is the same as but not limited to G105H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 106 of SEQ ID NO. 1, which is the same as but not limited to S106H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 107 of SEQ ID NO. 1, which is the same as but not limited to S107H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 109 of SEQ ID NO. 1, which is the same as but not limited to W109H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 113 of SEQ ID NO. 1, which is the same as but not limited to V113H. In some embodiments, the variable heavy chain includes a mutation corresponding to position 12 of SEQ ID NO. 1, which is the same as but not limited to V12K. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 20 of SEQ ID NO. 1, which is the same as but not limited to L20V. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 22 of SEQ ID NO. 1, which is the same as but not limited to C22A or C22K. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 38 of SEQ ID NO. 1, which is the same as but not limited to K38R. In some embodiments, the variable heavy chain comprises a mutation corresponding to position 76 of SEQ ID NO. 1, which is the same as but not limited to S76T.

[0217] In some embodiments, an antibody is provided comprising a variable heavy chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 1 and a variable light chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 2, provided, provided,

[0218] Variable light chain

[0219] Isoleucine corresponding to position 29 of sequence number 2,

[0220] Valine corresponding to position 34 of sequence number 2,

[0221] Glutamate corresponding to position 39 of sequence number 2,

[0222] Asparagine corresponding to position 58 of sequence number 2,

[0223] Glycine corresponding to position 96 of sequence number 2,

[0224] Proline corresponding to position 100 of sequence number 2,

[0225] Proline corresponding to position 101 of sequence number 2,

[0226] Threonine corresponding to position 102 of SEQ ID NO. 2,

[0227] Threonine corresponding to position 7 of SEQ ID NO. 2,

[0228] Serine corresponding to position 14 of sequence number 2,

[0229] Aspartate corresponding to position 17 of sequence number 2,

[0230] Leucine corresponding to position 42 of sequence number 2,

[0231] Lee Sin corresponding to position 44 of sequence number 2,

[0232] Leucine corresponding to position 51 of sequence number 2,

[0233] Alanine corresponding to position 72 of sequence number 2,

[0234] Arginine corresponding to position 79 of sequence number 2,

[0235] or includes any combination thereof.

[0236] In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises isoleucine corresponding to position 29 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises valine corresponding to position 34 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises glutamate corresponding to position 39 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises asparagine corresponding to position 58 of SEQ ID NO. 2.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises glycine corresponding to position 96 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises proline corresponding to position 100 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises proline corresponding to position 101 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises threonine corresponding to position 102 of SEQ ID NO. 2.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises threonine corresponding to position 7 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises a serine corresponding to position 14 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises aspartate corresponding to position 17 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises leucine corresponding to position 42 of SEQ ID NO. 2.In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises lysine corresponding to position 44 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises leucine corresponding to position 51 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises alanine corresponding to position 72 of SEQ ID NO. 2. In some embodiments, an antibody is provided comprising a variable heavy chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, provided that the variable light chain comprises arginine corresponding to position 79 of SEQ ID NO. 2.

[0237] In some embodiments, an antibody is provided comprising a variable heavy chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 1 and a variable light chain identical to at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of SEQ ID NO. 2, wherein the variable light chain is a mutation corresponding to position 24 of SEQ ID NO. 2 that is identical to but not limited to R24H, a mutation corresponding to position 25 of SEQ ID NO. 2 that is identical to but not limited to S25H, a mutation corresponding to position 26 of SEQ ID NO. 2 that is identical to but not limited to S26H, a mutation corresponding to position 27 of SEQ ID NO. 2 that is identical to but not limited to Q27H, and a mutation corresponding to S28H that is identical to but not limited to SEQ ID NO. A mutation corresponding to position 28 of 2, identical but not limited to V30H, a mutation corresponding to position 30 of SEQ ID NO. 2, identical but not limited to S32H, a mutation corresponding to position 32 of SEQ ID NO. 2, identical but not limited to N33H, a mutation corresponding to position 33 of SEQ ID NO. 2, identical but not limited to N35H, a mutation corresponding to position 35 of SEQ ID NO. 2, identical but not limited to T36H, a mutation corresponding to position 36 of SEQ ID NO. 2, identical but not limited to L38H, a mutation corresponding to position 38 of SEQ ID NO. 2, identical but not limited to K55H, a mutation corresponding to position 55 of SEQ ID NO. 2, identical but not limited to V56H, a mutation corresponding to position 56 of SEQ ID NO. 2, identical but not limited to S57H, a mutation corresponding to position 57 of SEQ ID NO. 2, identical but not limited to R59H, position of SEQ ID NO. 2 A mutation corresponding to 59, identical but not limited to F60H, a mutation corresponding to position 60 of SEQ ID NO. 2, identical but not limited to S61H,It includes a mutation corresponding to position 61 of SEQ ID NO. 2, a mutation corresponding to position 94 of SEQ ID NO. 2 that is the same as but not limited to F94H, a mutation corresponding to position 95 of SEQ ID NO. 2 that is the same as but not limited to Q95H, a mutation corresponding to position 97 of SEQ ID NO. 2 that is the same as but not limited to S97H, a mutation corresponding to position 99 of SEQ ID NO. 2 that is the same as but not limited to V99H, a mutation corresponding to position 41 of SEQ ID NO. 2 that is the same as but not limited to Y41F, a mutation corresponding to position 88 of SEQ ID NO. 2 that is the same as but not limited to L88V, a mutation corresponding to position 90 of SEQ ID NO. 2 that is the same as but not limited to I90V, or any combination thereof. In some embodiments, the variable light chain includes a mutation corresponding to position 24 of SEQ ID NO. 2 that is the same as but not limited to R24H. In some embodiments, the variable light chain comprises a mutation corresponding to position 25 of SEQ ID NO. 2, which is the same as but not limited to S25H. In some embodiments, the variable light chain comprises a mutation corresponding to position 26 of SEQ ID NO. 2, which is the same as but not limited to S26H. In some embodiments, the variable light chain comprises a mutation corresponding to position 27 of SEQ ID NO. 2, which is the same as but not limited to Q27H. In some embodiments, the variable light chain comprises a mutation corresponding to position 28 of SEQ ID NO. 2, which is the same as but not limited to S28H. In some embodiments, the variable light chain comprises a mutation corresponding to position 30 of SEQ ID NO. 2, which is the same as but not limited to V30H. In some embodiments, the variable light chain comprises a mutation corresponding to position 32 of SEQ ID NO. 2, which is the same as but not limited to S32H. In some embodiments, the variable light chain comprises a mutation corresponding to position 33 of SEQ ID NO. 2, which is the same as but not limited to N33H. In some embodiments, the variable light chain is the same as N35H but is not limited thereto,It includes a mutation corresponding to position 35 of SEQ ID NO. 2. In some embodiments, the variable light chain includes a mutation corresponding to position 36 of SEQ ID NO. 2, which is the same as but not limited to T36H. In some embodiments, the variable light chain includes a mutation corresponding to position 38 of SEQ ID NO. 2, which is the same as but not limited to L38H. In some embodiments, the variable light chain includes a mutation corresponding to position 55 of SEQ ID NO. 2, which is the same as but not limited to K55H. In some embodiments, the variable light chain includes a mutation corresponding to position 56 of SEQ ID NO. 2, which is the same as but not limited to V56H. In some embodiments, the variable light chain includes a mutation corresponding to position 57 of SEQ ID NO. 2, which is the same as but not limited to S57H. In some embodiments, the variable light chain includes a mutation corresponding to position 59 of SEQ ID NO. 2, which is the same as but not limited to R59H. In some embodiments, the variable light chain includes a mutation corresponding to position 60 of SEQ ID NO. 2, which is the same as but not limited to F60H. In some embodiments, the variable light chain comprises a mutation corresponding to position 61 of SEQ ID NO. 2, which is the same as but not limited to S61H. In some embodiments, the variable light chain comprises a mutation corresponding to position 94 of SEQ ID NO. 2, which is the same as but not limited to F94H. In some embodiments, the variable light chain comprises a mutation corresponding to position 95 of SEQ ID NO. 2, which is the same as but not limited to Q95H. In some embodiments, the variable light chain comprises a mutation corresponding to position 97 of SEQ ID NO. 2, which is the same as but not limited to S97H. In some embodiments, the variable light chain comprises a mutation corresponding to position 99 of SEQ ID NO. 2, which is the same as but not limited to V99H. In some embodiments, the variable light chain comprises a mutation corresponding to position 41 of SEQ ID NO. 2, which is the same as but not limited to Y41F. In some embodiments, the variable light chain comprises, which is the same as but not limited to L88V,Includes a mutation corresponding to position 88 of SEQ ID NO. 2. In some embodiments, the variable light chain includes a mutation corresponding to position 90 of SEQ ID NO. 2, which is the same as but not limited to I90V.

[0238] In some embodiments, the antibody comprises a variable heavy chain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2, and comprises at least one mutation (substitution) that is the same as but not limited to those provided herein, provided that the variable heavy chain comprises a mutation at position 26 that is the same as but not limited to G26H, a mutation at position 27 that is the same as but not limited to Y27H, a mutation at position 28 that is the same as but not limited to T28H, a mutation at position 29 that is the same as but not limited to F29H, and a mutation at position 29 that is the same as but not limited to T30H Mutation at 30, identical but not limited to S31H, mutation at position 31, identical but not limited to Y32H, mutation at position 32, identical but not limited to M34H, mutation at position 34, identical but not limited to I51H, mutation at position 51, identical but not limited to N59H, mutation at position 59, identical but not limited to A97H, mutation at position 97, identical but not limited to R98H, mutation at position 98, identical but not limited to G99H, mutation at position 99, identical but not limited to P101H, mutation at position 101, identical but not limited to D102H, mutation at position 102, identical but not limited to Y103H, mutation at position 103, identical but not limited to K108H, mutation at position 108, identical but not limited to Y110H, position Mutation at 110, mutation at position 111 identical to but not limited to F111H,A mutation at position 112 identical to, but not limited to, D112H; a mutation at position 40 identical to, but not limited to, R40A; a mutation at position 48 identical to, but not limited to, I48M; a mutation at position 67 identical to, but not limited to, K67R; a mutation at position 68 identical to, but not limited to, A68V; a mutation at position 70 identical to, but not limited to, L70M; a mutation at position 72 identical to, but not limited to, V72R; a mutation at position 74 identical to, but not limited to, K74T; a mutation at position 74 identical to, but not limited to, A79V; a mutation at position 79 identical to, but not limited to, Q82E; a mutation at position 82 identical to, but not limited to, T87R; a mutation at position 87 identical to, but not limited to, S91T; a mutation at position 91 identical to, but not limited to, S91T; and / or a variable light chain mutation at position 29 identical to, but not limited to, I29H, Mutation at position 34, identical but not limited to V34H; mutation at position 37, identical but not limited to Y37H; mutation at position 39, identical but not limited to E39H; mutation at position 58, identical but not limited to N58H; mutation at position 96, identical but not limited to G96H; mutation at position 100, identical but not limited to P100H; mutation at position 101, identical but not limited to P101H; mutation at position 102, identical but not limited to T102H; mutation at position 7, identical but not limited to T7S; mutation at position 14, identical but not limited to S14T; mutation at position 17, identical but not limited to D17Q; mutation at position 42, identical but not limited to L42Q; mutation at position 44, identical but not limited to K44RIt does not include a mutation at position 51 that is the same as, but not limited to, L51R, a mutation at position 72 that is the same as, but not limited to, A72S, or a mutation at position 79 that is the same as, but not limited to, R79K. In some embodiments, the variable heavy chain does not include a mutation at position 26 that is the same as, but not limited to, G26H. In some embodiments, the variable heavy chain does not include a mutation at position 27 that is the same as, but not limited to, Y27H. In some embodiments, the variable heavy chain does not include a mutation at position 28 that is the same as, but not limited to, T28H. In some embodiments, the variable heavy chain does not include a mutation at position 29 that is the same as, but not limited to, F29H. In some embodiments, the variable heavy chain does not include a mutation at position 30 that is the same as, but not limited to, T30H. In some embodiments, the variable heavy chain does not include a mutation at position 31 that is the same as, but not limited to, S31H. In some embodiments, the variable heavy chain does not contain a mutation at position 32, which is the same as but not limited to Y32H. In some embodiments, the variable heavy chain does not contain a mutation at position 34, which is the same as but not limited to M34H. In some embodiments, the variable heavy chain does not contain a mutation at position 51, which is the same as but not limited to I51H. In some embodiments, the variable heavy chain does not contain a mutation at position 59, which is the same as but not limited to N59H. In some embodiments, the variable heavy chain does not contain a mutation at position 97, which is the same as but not limited to A97H. In some embodiments, the variable heavy chain does not contain a mutation at position 98, which is the same as but not limited to R98H. In some embodiments, the variable heavy chain does not contain a mutation at position 99, which is the same as but not limited to G99H. In some embodiments, the variable heavy chain is the same as but not limited to P101H,It does not include a mutation at position 101. In some embodiments, the variable heavy chain does not include a mutation at position 102, which is the same as but not limited to D102H. In some embodiments, the variable heavy chain does not include a mutation at position 103, which is the same as but not limited to Y103H. In some embodiments, the variable heavy chain does not include a mutation at position 108, which is the same as but not limited to K108H. In some embodiments, the variable heavy chain does not include a mutation at position 110, which is the same as but not limited to Y110H. In some embodiments, the variable heavy chain does not include a mutation at position 111, which is the same as but not limited to F111H. In some embodiments, the variable heavy chain does not include a mutation at position 112, which is the same as but not limited to D112H. In some embodiments, the variable heavy chain does not include a mutation at position 40, which is the same as but not limited to R40A. In some embodiments, the variable heavy chain does not contain a mutation at position 48, which is the same as but not limited to I48M. In some embodiments, the variable heavy chain does not contain a mutation at position 67, which is the same as but not limited to K67R. In some embodiments, the variable heavy chain does not contain a mutation at position 68, which is the same as but not limited to A68V. In some embodiments, the variable heavy chain does not contain a mutation at position 70, which is the same as but not limited to L70M. In some embodiments, the variable heavy chain does not contain a mutation at position 72, which is the same as but not limited to V72R. In some embodiments, the variable heavy chain does not contain a mutation at position 74, which is the same as but not limited to K74T. In some embodiments, the variable heavy chain does not contain a mutation at position 79, which is the same as but not limited to A79V. In some embodiments, the variable heavy chain is the same as but not limited to Q82E,It does not include a mutation at position 82. In some embodiments, the variable heavy chain does not include a mutation at position 87, which is the same as but not limited to T87R. In some embodiments, the variable heavy chain does not include a mutation at position 91, which is the same as but not limited to S91T. In some embodiments, the variable light chain does not include a mutation at position 29, which is the same as but not limited to I29H. In some embodiments, the variable light chain does not include a mutation at position 34, which is the same as but not limited to V34H. In some embodiments, the variable light chain does not include a mutation at position 37, which is the same as but not limited to Y37H. In some embodiments, the variable light chain does not include a mutation at position 39, which is the same as but not limited to E39H. In some embodiments, the variable light chain does not include a mutation at position 58, which is the same as but not limited to N58H. In some embodiments, the variable light chain does not include a mutation at position 96, which is the same as but not limited to G96H. In some embodiments, the variable light chain does not contain a mutation at position 100, which is the same as but not limited to P100H. In some embodiments, the variable light chain does not contain a mutation at position 101, which is the same as but not limited to P101H. In some embodiments, the variable light chain does not contain a mutation at position 102, which is the same as but not limited to T102H. In some embodiments, the variable light chain does not contain a mutation at position 7, which is the same as but not limited to T7S. In some embodiments, the variable light chain does not contain a mutation at position 14, which is the same as but not limited to S14T. In some embodiments, the variable light chain does not contain a mutation at position 17, which is the same as but not limited to D17Q. In some embodiments, the variable light chain is the same as but not limited to L42Q,It does not include a mutation at position 42. In some embodiments, the variable light chain does not include a mutation at position 44, which is the same as but not limited to K44R. In some embodiments, the variable light chain does not include a mutation at position 51, which is the same as but not limited to L51R. In some embodiments, the variable light chain does not include a mutation at position 72, which is the same as but not limited to A72S. In some embodiments, the variable light chain does not include a mutation at position 79, which is the same as but not limited to R79K.

[0239] In some embodiments, the variable heavy chain comprises an amino acid sequence and a W33H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 10.

[0240] In some embodiments, the variable heavy chain comprises an amino acid sequence and an E50H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 12.

[0241] In some embodiments, the variable heavy chain comprises an amino acid sequence and an N52H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 14.

[0242] In some embodiments, the variable heavy chain comprises an amino acid sequence and a P53H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 15.

[0243] In some embodiments, the variable heavy chain comprises an amino acid sequence and an S54H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 16.

[0244] In some embodiments, the variable heavy chain comprises an amino acid sequence and an N55H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 17.

[0245] In some embodiments, the variable heavy chain comprises an amino acid sequence and a G56H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 18.

[0246] In some embodiments, the variable heavy chain comprises an amino acid sequence and an R57H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 19.

[0247] In some embodiments, the variable heavy chain comprises an amino acid sequence and a T58H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 20.

[0248] In some embodiments, the variable heavy chain comprises an amino acid sequence and a Y60H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 22.

[0249] In some embodiments, the variable heavy chain comprises an amino acid sequence and an N61H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 23.

[0250] In some embodiments, the variable heavy chain comprises an amino acid sequence and a Q62H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 24.

[0251] In some embodiments, the variable heavy chain comprises an amino acid sequence and a K63H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 25.

[0252] In some embodiments, the variable heavy chain comprises an amino acid sequence and an F64H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 26.

[0253] In some embodiments, the variable heavy chain comprises an amino acid sequence and a Q65H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 27.

[0254] In some embodiments, the variable heavy chain comprises an amino acid sequence and a G66H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 28.

[0255] In some embodiments, the variable heavy chain comprises an amino acid sequence and an F96H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 29.

[0256] In some embodiments, the variable heavy chain comprises an amino acid sequence and an R100H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 33.

[0257] In some embodiments, the variable heavy chain comprises an amino acid sequence and a Y104H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 37.

[0258] In some embodiments, the variable heavy chain comprises an amino acid sequence and a G105H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 38.

[0259] In some embodiments, the variable heavy chain comprises an amino acid sequence and an S106H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 39.

[0260] In some embodiments, the variable heavy chain comprises an amino acid sequence and an S107H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 40.

[0261] In some embodiments, the variable heavy chain comprises an amino acid sequence and a W109H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 42.

[0262] In some embodiments, the variable heavy chain comprises an amino acid sequence and a V113H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 46.

[0263] In some embodiments, the variable heavy chain comprises an amino acid sequence and a V12K mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 77.

[0264] In some embodiments, the variable heavy chain comprises an amino acid sequence and an L20V mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 78.

[0265] In some embodiments, the variable heavy chain comprises an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1 and a C22A or C22K mutation. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 79 or 80.

[0266] In some embodiments, the variable heavy chain comprises an amino acid sequence and a K38R mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 81.

[0267] In some embodiments, the variable heavy chain comprises an amino acid sequence and an S76T mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 1. In some embodiments, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 89.

[0268] In some embodiments, the variable light chain comprises an amino acid sequence and an R24H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 47.

[0269] In some embodiments, the variable light chain comprises an amino acid sequence and an S25H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 48.

[0270] In some embodiments, the variable light chain comprises an amino acid sequence and an S26H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 49.

[0271] In some embodiments, the variable light chain comprises an amino acid sequence and a Q27H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 50.

[0272] In some embodiments, the variable light chain comprises an amino acid sequence and an S28H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 51.

[0273] In some embodiments, the variable light chain comprises an amino acid sequence and a V30H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 53.

[0274] In some embodiments, the variable light chain comprises an amino acid sequence and an S32H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 54.

[0275] In some embodiments, the variable light chain comprises an amino acid sequence and an N33H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 55.

[0276] In some embodiments, the variable light chain comprises an amino acid sequence and an N35H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 57.

[0277] In some embodiments, the variable light chain comprises an amino acid sequence and a T36H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 58.

[0278] In some embodiments, the variable light chain comprises an amino acid sequence and an L38H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 60.

[0279] In some embodiments, the variable light chain comprises an amino acid sequence and a K55H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 62.

[0280] In some embodiments, the variable light chain comprises an amino acid sequence and a V56H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 63.

[0281] In some embodiments, the variable light chain comprises an amino acid sequence and an S57H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 64.

[0282] In some embodiments, the variable light chain comprises an amino acid sequence and an R59H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 66.

[0283] In some embodiments, the variable light chain comprises an amino acid sequence and an F60H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 67.

[0284] In some embodiments, the variable light chain comprises an amino acid sequence and an S61H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 68.

[0285] In some embodiments, the variable light chain comprises an amino acid sequence and an F94H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 69.

[0286] In some embodiments, the variable light chain comprises an amino acid sequence and a Q95H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 70.

[0287] In some embodiments, the variable light chain comprises an amino acid sequence and an S97H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 72.

[0288] In some embodiments, the variable light chain comprises an amino acid sequence and a V99H mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 73.

[0289] In some embodiments, the variable light chain comprises an amino acid sequence and a Y41F mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 97.

[0290] In some embodiments, the variable light chain comprises an amino acid sequence and an L88V mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 103.

[0291] In some embodiments, the variable light chain comprises an amino acid sequence and an I90V mutation that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO. 2. In some embodiments, the variable light chain comprises the amino acid sequence of SEQ ID NO. 104.

[0292] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 10.

[0293] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 12.

[0294] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 14.

[0295] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 15.

[0296] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 16.

[0297] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 17.

[0298] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 18.

[0299] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 19.

[0300] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 20.

[0301] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 22.

[0302] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 23.

[0303] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 24.

[0304] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 25.

[0305] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 26.

[0306] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 27.

[0307] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 28.

[0308] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 29.

[0309] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 33.

[0310] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 37.

[0311] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 38.

[0312] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 39.

[0313] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 40.

[0314] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 42.

[0315] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 46.

[0316] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 77.

[0317] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 78.

[0318] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 79.

[0319] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 80.

[0320] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 81.

[0321] In some embodiments, the antibody comprises a variable light chain having the amino acid sequence of SEQ ID NO. 2 and a variable heavy chain having the amino acid sequence of SEQ ID NO. 89.

[0322] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 47.

[0323] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 48.

[0324] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 49.

[0325] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 50.

[0326] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 51.

[0327] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 53.

[0328] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 54.

[0329] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 55.

[0330] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 57.

[0331] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 58.

[0332] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 60.

[0333] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 62.

[0334] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 63.

[0335] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 64.

[0336] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 66.

[0337] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 67.

[0338] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 68.

[0339] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 69.

[0340] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 70.

[0341] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 72.

[0342] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 73.

[0343] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 97.

[0344] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 103.

[0345] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 104.

[0346] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 15 and a variable light chain having the amino acid sequence of SEQ ID NO. 2.

[0347] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 17 and a variable light chain having the amino acid sequence of SEQ ID NO. 2.

[0348] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 70.

[0349] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 72.

[0350] In some embodiments, the antibody comprises a variable heavy chain having the amino acid sequence of SEQ ID NO. 1 and a variable light chain having the amino acid sequence of SEQ ID NO. 73.

[0351] In addition to these specific combinations, an arbitrary V H Peptides and V L Peptides can be combined with each other.

[0352] Additionally, as provided herein, the antibody may be a multi-specific antibody in that the antibody has multiple binding regions that target the same protein at different proteins or different epitopes. In some embodiments, the antibody is a bispecific antibody.

[0353] As provided herein, different peptides (V) described herein H or V L ) can be linked to a peptide linker or not linked to a peptide linker and instead linked for a continuous sequence. In some embodiments, the peptide linker is (GGGGS) n (Sequence No. 105), (GGGGA) n It comprises a sequence of (SEQ ID NO. 106) or any combination thereof, wherein each n is independently 1 to 5. The linked peptide format is V H -ZV L or V L -ZV H It can be represented by the chemical formula, where Z is a peptide linker. In some embodiments, Z is (GGGGS) n (Sequence No. 105), (GGGGA) n (Sequence No. 106) or any combination thereof, wherein each n is independently 1 to 5. Such antibodies may be referred to as scFv format.

[0354] 치료 방법

[0355] Treatment methods are provided at this hospital.

[0356] In some embodiments, a method for treating thyroid-associated eye disease (TAO) or its symptoms or reducing its severity is provided. In some embodiments, a method for treating thyroid-associated eye disease (TAO) or its symptoms or reducing its severity comprises administering any antibody disclosed herein to a subject. In some embodiments, a method for reducing proptosis in the eye of a subject having thyroid-associated eye disease (TAO) is provided. In some embodiments, a method for reducing proptosis in the eye of a subject having thyroid-associated eye disease (TAO) comprises administering any antibody disclosed herein to a subject.

[0357] In some embodiments, a method for treating thyroid eye disease (TED) in a subject is provided, which comprises administering any antibody disclosed herein to the subject. In some embodiments, the subject has active thyroid eye disease or chronic thyroid eye disease. Active thyroid eye disease refers to a subject diagnosed with thyroid eye disease within 1 to 2 years of administration of the therapeutic agent and having a CAS greater than 1. Chronic thyroid eye disease generally refers to an initial diagnosis of thyroid eye disease 2 to 10 years prior to administration of the therapeutic agent, in which the Clinical Activity Score (CAS) in both eyes is 1 or less for at least 1 year prior to screening, or in which there is no progression of proptosis, no progression of diplopia, and no new inflammatory TED or TAO symptoms for 1 year prior to screening.

[0358] In some embodiments, a method for reducing the clinical activity score (CAS) of thyroid-associated eye disease (TAO) in a subject is provided, which comprises administering any antibody disclosed herein to the subject.

[0359] In some embodiments, a method is provided for a) reducing proptosis by at least 2 mm and b) reducing the Clinical Activity Score (CAS) in a subject having thyroid-associated eye disease (TAO), which comprises administering any antibody disclosed herein to the subject. In some embodiments, proptosis is reduced by at least 2 mm. In some embodiments, proptosis is reduced by at least 3 mm. In some embodiments, proptosis is reduced by at least 4 mm. In some embodiments, 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) point. In some embodiments, the subject's Clinical Activity Score (CAS) is reduced to zero (0) point.

[0360] In some embodiments, a method for treating thyroid-associated eye disease (TAO) in a subject or reducing its severity is provided, comprising administering any antibody disclosed herein to the subject, wherein treatment with said antibody (i) reduces proptosis of one eyeball by at least 2 mm, (ii) does not entail worsening of the opposite eyeball (or opposite eye) by more than 2 mm, and (iii) reduces the CAS of said subject to one (1) or zero (0).

[0361] In some embodiments, a method for improving the quality of life of a subject having thyroid-associated eye disease (TAO, also known as Graves' eye disease / Graves' orbital disease) is provided, which comprises administering any of the antibodies disclosed herein to the subject.

[0362] In some embodiments, quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or by either the visual function or appearance subscales thereof. In some embodiments, treatment results in an improvement of 8 points or more in the GO-QoL. In some embodiments, treatment results in an improvement in the function subscale of the GO-QoL. In some embodiments, treatment results in an improvement in the appearance subscale of the GO-QoL.

[0363] In some embodiments, a method for treating diplopia or reducing its severity in a subject having thyroid-associated eye disease (TAO) is provided, comprising administering any antibody disclosed herein to the subject. In some embodiments, the diplopia is persistent diplopia. In some embodiments, the diplopia is non-persistent diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, the improvement in diplopia or the reduction in its severity is maintained for at least 20 weeks after discontinuation of antibody administration. In some embodiments, the improvement in diplopia or the reduction in its severity is maintained for at least 50 weeks after discontinuation of antibody administration.

[0364] In some embodiments, the antibody is administered as a first dose at a dosage of about 1 mg / kg to about 20 mg / kg, and then the same dose is administered as a subsequent dose. In some embodiments, doses are administered at 1, 3, 5, 10, 15, or 20 mg / kg every 3, 4, 5, 6, 7, 8, or 12 weeks. In some embodiments, the antibody is administered every 3 weeks. In some embodiments, a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 doses of the antibody are administered. In some embodiments, a total of 5 doses are administered. In some embodiments, a total of 8 doses are administered. In some embodiments, subsequent doses after the initial dose are administered every 3 weeks for at least 12, 15, 18, or 21 weeks. In some embodiments, the antibody or its antigen-binding fragment is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a diabadi, a single-chain antibody, a multi-specific antibody, Fab, Fab', F(ab')2, Fv, or scFv.

[0365] In some embodiments, a method is provided in which an antibody or an antigen-binding fragment thereof is administered into a pharmaceutical composition further comprising a pharmaceutically acceptable diluent or excipient or carrier.

[0366] In some embodiments, a method is provided in which the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. In some embodiments, a method is provided in which 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.

[0367] In some embodiments, a method is provided in which an antibody or an antigen-binding fragment thereof is administered directly into the eye, the anterior chamber of the eye, the vitreous cavity of the eye, the supracoracial space, or the posterior orbital sinus. In some embodiments, a method is provided in which an antibody or an antigen-binding fragment thereof is administered via injection. In some embodiments, an injection method is provided in which the injection is an intravitreal injection, an intraorbital injection, a posterior orbital injection, a supracoracial injection, or an anterior chamber injection.

[0368] In some embodiments, a method for increasing the internalization of IGF-1R in cells is provided, the method comprises contacting the cells with the antibody provided herein. In some embodiments, the contact method comprises administering the antibody provided herein to a subject. In some embodiments, a method is provided in which a subject has thyroid eye disease (TED) or is at risk thereof.

[0369] In some embodiments, a method for inhibiting IGF-1-stimulated receptor phosphorylation on cells is provided, the method comprising contacting the cells with the antibody provided herein. In some embodiments, a method is provided in which contacting comprises administering the antibody provided herein to a subject. In some embodiments, a method is provided in which the subject has thyroid eye disease (TED) or is at risk thereof. In some embodiments, the antibody has an IC of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less 50 A method having is provided. In some embodiments, IC 50 A method is provided for measuring in an in vitro assay that is the same as, but not limited to, the assay provided herein. In some embodiments, a method is provided in which the cells are A549 cells or HOCF cells.

[0370] In some embodiments, a method for treating thyroid eye disease (TED) in a subject is provided, the method comprises administering the antibody provided herein, wherein the antibody has a serum concentration in the subject of 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 after at least 1, 2, or 3 weeks after administration. In some embodiments, the antibody is administered intravenously. In some embodiments, the antibody is administered subcutaneously.

[0371] In some embodiments, a method is provided for inhibiting IGF-1-induced receptor autophosphorylation in cells by at least 95%, 96%, 97%, 98%, 99%, or 100%, the method comprises contacting the cells with the antibody provided herein. In some embodiments, a method is provided in which the inhibition of IGF-1-induced receptor autophosphorylation is measured in comparison to receptor autophosphorylation induced in the absence of the antibody. In some embodiments, a method is provided in which contacting comprises administering the antibody comprising the above to a subject. In some embodiments, a method is provided in which the subject has thyroid eye disease (TED) or is at risk thereof. In some embodiments, a method is provided for inhibiting IGF-1-induced receptor autophosphorylation in a subject requiring at least 95%, 96%, 97%, 98%, 99%, or 100%, the method comprises administering the antibody provided herein to the subject. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof.

[0372] 약제학적 조성물

[0373] The present invention provides pharmaceutical or sterile compositions comprising the antibodies provided herein. A pharmaceutical or sterile composition of an anti-IGF-1R antibody or an antigen-binding fragment thereof may be mixed with a pharmaceutically acceptable carrier or excipient to produce a pharmaceutical composition. For example, literature [ Remington's Pharmaceutical Sciences and US Pharmacopoeia: National Formulary See Mack Publishing Company, Easton, PA (1984).

[0374] Formulations of therapeutic and diagnostic agents may be prepared by mixing acceptable carriers, excipients, or stabilizers in the form of, for example, lyophilized powders, slurries, aqueous solutions, or suspensions (e.g., literature [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], [Weiner and Kotkoskie (2000) Excipient Toxicity and Safety [See Marcel Dekker, Inc., New York, NY]). In some embodiments, the antibody is diluted to an appropriate concentration in a sodium acetate solution of pH 5 to 6, and NaCl or sucrose is added for isotonicity. Additional agents such as polysorbate 20 or polysorbate 80 may be added to improve stability.

[0375] The toxicity and therapeutic efficacy of an antibody composition administered alone or in combination with other agents can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, LD 50 (lethal dose for 50% of the population) and ED 50 It is intended to determine (the therapeutically effective dose in 50% of the population). The dose ratio between toxic effects and therapeutic effects is the therapeutic index (LD50). 50 / ED 50 ...is. Antibodies exhibiting a high therapeutic index in a specific embodiment are preferred. Data obtained from these cell culture assays and animal studies can be used to formulate dosage ranges for human use. The dosage of these compounds is preferably ED with little to no toxicity. 50 It is within the range of circulating concentrations including. The dosage may vary within this range depending on the dosage form and route of administration used.

[0376] In some embodiments, the antibody, or its antigen-binding fragment, or its pharmaceutical composition provided herein is administered to a subject in accordance with the literature [Physicians' Desk Reference 2003 (Thomson Healthcare; 57th edition (November 1, 2002))].

[0377] The method of administration may vary. Suitable routes of administration include oral, rectal, transmucosal, intestinal, parenteral; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, cutaneous, transcutaneous, or intra-arterial.

[0378] In some embodiments, a pharmaceutical composition comprising an antibody or an antigen-binding fragment thereof may be administered by an invasive route, such as injection. In some embodiments, the pharmaceutical composition is administered intravenously, subcutaneously, intramuscularly, intra-arterially, intra-articularly (e.g., arthritic

[0379] In some embodiments, an antibody or an antigen-binding fragment thereof, or a pharmaceutical composition thereof is administered directly to the eye, the anterior chamber of the eye, the vitreous cavity of the eye, the supracoracial space, or the retroorbital sinus. In some embodiments, administration to the eye, the anterior chamber of the eye, the vitreous cavity of the eye, the supracoracial space, or the retroorbital sinus is by injection. In some embodiments, the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a supracoracial injection, or an anterior chamber 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 retroorbital injection. In some embodiments, the injection is a supracoracial injection. In some embodiments, the injection is an anterior chamber injection.

[0380] In some embodiments, the anti-IGF-1R antibody or its antigen-binding fragment is administered in combination with at least one additional therapeutic agent, which is the same as but not limited to any therapeutic agent used to treat thyroid eye disease. For example, in some embodiments, the anti-IGF-1R antibody or its antigen-binding fragment is administered in combination with at least one additional therapeutic agent, which is the same as but not limited to a therapeutic agent used to treat thyroid eye disease or related conditions. Examples of such treatments and therapeutic agents include anti-thyroid drugs, diabetes drugs, beta-blockers, propylthiouracil, methimazole, propranolol, atenolol, metoprolol, nadolol, corticosteroids, metformin, sulfonylureas, meglitinides, thiazolidinediones, DPP-4 inhibitors, GLP-1 receptor agonists, SGLT2 inhibitors, regular insulin, insulin aspart, insulin glulisin, insulin lispro, insulin isophanate, insulin degludec, insulin detemir, insulin glargine, acervose, miglitol, acebutolol, atenolol, betaxolol, bisoprolol, cartelol, carvedilol, esmolol, labetalol, metoprolol, nadolol, nebivolol, fenbutolol, pindolol, propranolol, sotalol, timolol, tomolol eye drops. It includes, but is not limited to, sitagliptin, saxagliptin, linagliptin, alogliptin, dulaglutide, exenatide, semaglutide, liraglutide, lixisenatide, canagliflozin, dapagliflozin, empagliflozin, or any combination thereof.

[0381] The composition may be administered using a medical device known in the art. For example, the pharmaceutical composition of the present invention may be administered by injection using a subcutaneous injection needle, for example, a pre-filled syringe or an auto-injector.

[0382] The pharmaceutical composition may also be administered using a needle subcutaneous injection device, such as the device disclosed in U.S. Patent No. 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.

[0383] Pharmaceutical compositions may also be administered by infusion. Examples of known implants and modules for administering pharmaceutical compositions include U.S. Patent No. 4,487,603 disclosing an implantable micro-infusion pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,447,233 disclosing a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224 disclosing a variable flow implantable infusion device for continuous drug delivery; and U.S. Patent No. 4,439,196 disclosing an osmotic drug delivery system having multiple chamber compartments. Many other such implants, delivery systems, and modules are known to those skilled in the art.

[0384] Alternatively, humans may administer antibodies locally rather than systemically, for example, through the direct injection of antibodies into arthritic joints or pathogen-induced lesions characterized by immunopathology, often in depot or sustained-release formulations. Furthermore, humans may administer antibodies via targeted drug delivery systems, for example, as liposomes coated with tissue-specific antibodies targeting arthritic joints or pathogen-induced lesions characterized by immunopathology. The liposomes will be targeted to the affected tissue and selectively absorbed by it.

[0385] The administration regimen depends on several factors, including the serum or tissue turnover of therapeutic antibodies, the level of symptoms, the immunogenicity of the therapeutic antibodies, and the accessibility of target cells within the biological matrix. Preferably, the administration regimen delivers sufficient therapeutic antibodies to bring about improvement in the target disease state while simultaneously minimizing unintended side effects. Thus, the amount of biopharmaceutical delivered depends in part on the specific therapeutic antibody and the severity of the condition being treated. Guidelines are available for selecting the appropriate dose of therapeutic antibodies (e.g., literature [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. [Refer to 343:1594-1602]).

[0386] The determination of the appropriate dose is made by a clinician, for example, using parameters or factors known or suspected in the art to influence treatment. Generally, the dose is started at a level slightly lower than the optimal dose and then increased in small increments until the desired or optimal effect is achieved, in case of any adverse side effects. Important diagnostic measurements include, for example, the indications of inflammation or the measurement of levels of inflammatory cytokines produced. Generally, it is preferable that the biopharmaceutical to be used be derived from the same species as the animal being targeted for treatment, thereby minimizing any immune response to the reagent. For human subjects, for example, chimeric, humanized, and fully human antibodies may be preferred.

[0387] The antibody or its antigen-binding fragment provided herein may be administered by continuous infusion, or at dosages such as, for example, daily, 1 to 7 times per week, weekly, bi-weekly, monthly, bi-monthly, quarterly, semi-annually, annually, etc. Doses may be provided, for example, intravenously, subcutaneously, topically, orally, intranasally, rectally, intramuscularly, intracerebrally, intraspinally, or by inhalation. In some embodiments, the antibody is administered every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, or every 8 weeks. In some embodiments, the antibody is administered every 4 weeks. In some embodiments, the antibody is administered every 5 weeks. In some embodiments, the antibody is administered every 7 weeks. In some embodiments, the antibody is administered every 6 weeks. In some embodiments, the antibody is administered every 8 weeks. In some embodiments, the antibody is administered for at least 21 to 52 weeks or longer. In some embodiments, the antibody is administered according to this schedule for at least 21 weeks. In some embodiments, the antibody is administered according to this schedule for at least 24 weeks. In some embodiments, the antibody is administered according to this schedule for at least 32 weeks. In some embodiments, the antibody is administered according to this schedule for at least 36 weeks. In some embodiments, the antibody is administered according to this schedule for at least 40 weeks. In some embodiments, the antibody is administered according to this schedule for at least 42 weeks. In some embodiments, the antibody is administered once (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered twice (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered three times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered four times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered five times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered six times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered seven times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered eight times (e.g., by infusion or subcutaneous injection).In some embodiments, the antibody is administered 9 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 10 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 11 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 12 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 13 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 14 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 15 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 16 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 17 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 18 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 19 times (e.g., by infusion or subcutaneous injection). In some embodiments, the antibody is administered 20 times (e.g., by infusion or subcutaneous injection). If the antibody is administered more than once, it may be administered according to a schedule such as the schedule provided herein.

[0388] The total weekly dose may be as provided herein. In some embodiments, the total weekly dose is at least 0.05 μg / kg body weight, more generally at least 0.2 μg / kg, 0.5 μg / kg, 1 μg / kg, 10 μg / kg, 100 μg / kg, 0.25 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 (e.g., the literature [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. [See 52:133-144]). Doses may also be provided to achieve a predetermined target concentration of antibody in the subject's serum, such as 0.1, 0.3, 1, 3, 10, 30, 100, 300 μg / ml or higher.

[0389] In some embodiments, the antibody has a serum concentration in the subject of 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 after at least 1, 2 or 3 weeks of administration.

[0390] In some embodiments, a dose of 3, 5, 10, or 20 mg / kg IV is administered, and in some embodiments, the dose is 133 μg / mL C after about 5 weeks. min It is used to provide. In some embodiments, the dose of antibody administered is 102 μg / mL C after 6 weeks. min It provides. In some embodiments, the dose of the antibody is as provided herein as a loading dose, such as 3, 5, 10, or 20 mg / kg, and subsequent doses are equal to or lower than this. In some embodiments, the antibody is at least or about 100 μg / mL of C min It is administered as provided herein in a dose to achieve.

[0391] As used herein, "inhibit," "treat," or "treat" include the delay of the onset of symptoms associated with a disorder and / or the reduction of the severity of such symptoms. These terms further include improving existing uncontrolled or unwanted symptoms, preventing additional symptoms, and improving or preventing the underlying causes of such symptoms. Accordingly, these terms indicate that a beneficial outcome has been conferred upon a vertebrate subject having a disorder, disease, or symptoms, or having the potential to develop such disorder, disease, or symptoms.

[0392] As used herein, the terms 'therapeutic effective dose,' 'therapeutic effective dose,' and 'effective dose' refer to the amount of an antibody or its antigen-binding fragment effective in causing one or more symptoms of a disease or condition or a measurable improvement in the progression of such disease or condition when administered to a cell, tissue, or subject, either alone or in combination with additional therapeutic agents. The therapeutic effective dose further refers to an amount of binding compound sufficient to cause at least partial improvement of symptoms, e.g., treatment, cure, prevention, or improvement of a related medical condition, or an increase in the rate of treatment, cure, prevention, or improvement of such condition. When applied to an individual active ingredient administered alone, the therapeutic effective dose refers to that ingredient alone. When applied to a combination, the therapeutic effective dose refers to the combined amount of active ingredients that produce a therapeutic effect, regardless of whether they are administered in combination, sequentially, or simultaneously. An effective dose of the therapeutic agent will cause an improvement in diagnostic measurements or parameters of at least 10%, generally at least 20%, preferably at least about 30%, more preferably at least 40%, and most preferably at least 50%. The effective dose may also cause an improvement in subjective measurements where subjective measurements are used to assess disease severity. In some embodiments, if the amount is an amount that can be used to treat or improve the condition provided herein, it is a therapeutically effective dose.

[0393] The term 'subject' as used throughout includes any organism, such as animals including mammals (e.g., rats, mice, dogs, cats, rabbits), and, for example, humans. 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 identified as requiring treatment for a condition to be treated and receiving treatment with a specific intention to treat such condition. The condition may be, for example, any condition described herein.

[0394] On the other hand, since isolated antibodies bind to epitopes on the IGF-1R protein or other proteins described herein and exhibit IGF-1R inhibition or therapeutic activity in vitro and / or in vivo, any antibody or antigen-binding fragment thereof capable of inhibiting IGF-1R function is suitable as a therapeutic agent for treating IGF-1R-associated conditions in humans and animals. Such conditions include thyroid eye disease. Accordingly, a method for treating such conditions is also provided, wherein the method comprises administering an antibody or an antigen-binding fragment thereof to a subject having such conditions.

[0395] In some embodiments, the method comprises administering a therapeutically or prophylactically effective amount of one or more monoclonal antibodies or antigen-binding fragments of antibodies described herein to a susceptible subject or a subject exhibiting a condition known or suspected to cause an observed pathology involving IGF-1R. Any active form of the antibody may be administered, which includes, but is not limited to, scFV, Fab and F(ab')2 fragments, and other forms of the antibodies provided herein.

[0396] As used herein, IGF-1R-associated pathology refers to conditions induced by the regulation of IGF-1R. These conditions include, but are not limited to, thyroid eye disease and other conditions provided herein.

[0397] The antibody used in some embodiments is compatible with the recipient species so as not to cause an unacceptably short cyclic half-life of the immune response to the mAb or not to induce an immune response to the mAb in the subject.

[0398] Treatment of the individual may include the administration of a therapeutically effective amount of the antibody described herein. The antibody may be provided in a kit such as those provided herein. The antibody may be used or administered alone or in combination with other therapeutic agents, analgesics, or diagnostic agents as provided herein. When providing a patient with an antibody or a fragment thereof capable of binding to IGF-1R, or an antibody capable of protecting against IGF-1R pathology in the recipient patient, the dosage of the administered agent will vary depending on factors such as the patient's age, weight, height, sex, general medical condition, previous medical history, etc.

[0399] Antibodies capable of treating conditions associated with IGF-1R activity or used to treat IGF-1R-related pathologies are intended to be provided to subjects in an amount sufficient to influence the reduction, resolution, or improvement of IGF-1R-related symptoms or pathologies. Such pathologies include thyroid eye disease, etc.

[0400] Accordingly, in some embodiments, a method for treating a subject having an IGF-1R-mediated disorder is provided. In some embodiments, the method comprises administering a pharmaceutical composition comprising the antibody provided herein or an antigen-binding fragment thereof. In some embodiments, the disorder is thyroid eye disease. The antibody provided herein or an antigen-binding fragment thereof may be administered together with other therapeutic agents. These may be administered simultaneously or sequentially.

[0401] In some embodiments, an antibody or an antigen-binding fragment thereof may be used to treat thyroid eye disease. In some embodiments, an antibody or an antigen-binding fragment thereof may be used to treat thyroid-associated eye disease (TAO) or its symptoms or to reduce its severity.

[0402] In some embodiments, a method or use for reducing proptosis in the eye of a subject having thyroid-associated eye disease (TAO) is provided.

[0403] In some embodiments, the subject is a subject who has previously been treated with an antibody different from that provided herein.

[0404] In some embodiments, a method or use for a Clinical Activity Score (CAS) is provided for subjects who have or are suspected of having thyroid-associated eye disease (TAO).

[0405] In some embodiments, a method or use is provided for reducing proptosis by at least 2 mm in subjects with thyroid-associated eye disease (TAO) and reducing the clinical activity score (CAS).

[0406] The term Clinical Activity Score (CAS) used in this document is Table 3 It refers to a protocol that is described and scored according to [the protocol]. According to this protocol, the following Table 3 One point is given for the presence of each parameter evaluated in. The sum of all scores defines clinical activity and provides CAS, where 0 or 1 constitutes inactive disease and 7 constitutes severe active ophthalmopathy.

[0407]

[0408] Table 3As provided, CAS consists of seven components: spontaneous retrobulbar pain, pain during ocular movement (upward, left-right, and downward gaze), conjunctival redness, eyelid redness, conjunctival edema, swelling of the melanocytic / lumbrical folds, and eyelid swelling. Each component is scored as present (1 point) or absent (0 points). The score for each efficacy evaluation is the sum of all present items; it provides a range of 0 to 7 points, where 0 or 1 point constitutes inactive disease and 7 points constitutes severe active ophthalmopathy. A change of more than 2 points is considered clinically significant.

[0409] Item 1, Spontaneous orbital pain may be pain or pressure felt above or behind the eye. This pain can be caused by increased intraorbital pressure as orbital tissue volume increases through the excessive synthesis of the extracellular matrix, fluid accumulation, and cellular infiltration and expansion. Item 2, Gaze-induced orbital pain may be pain in the eye when looking up, down, or to the side, or attempting to look; that is, pain during upward, downward, or lateral eye movements or when attempting eye movements. This type of pain may arise from the stretching of inflamed muscle(s) and may occur particularly when attempting to gaze upward. 'Stretch pain' cannot be triggered by pressing on the eye with a finger, as would be expected in the case of signs of increased intraorbital pressure. Both types of pain may decrease after anti-inflammatory treatment. Therefore, this type of pain is considered to be directly related to autoimmune inflammation within the orbit and is thus useful for evaluating TAO activity.

[0410] Swelling in TAO Table 3Item 6 is observed as conjunctival edema (edema of the conjunctiva) and swelling of the pylorus and / or lunula. Both are signs of TAO activity. Swollen eyelids may be caused by edema, fat prolapse through the orbital septum, or fibrotic degeneration. In addition to swelling, other symptoms indicating active TAO include redness and / or pain of the conjunctiva, eyelids, pylorus and / or lunula.

[0411] In some embodiments, the exophthalmos of the subject being treated is reduced by at least 2 mm. In some embodiments, the exophthalmos of the subject being treated is reduced by at least 3 mm. In some embodiments, the exophthalmos of the subject being treated is reduced by at least 4 mm.

[0412] In some embodiments, the clinical activity score (CAS) of the subject being treated is reduced by at least 2 points. In some embodiments, the clinical activity score (CAS) of the subject is reduced by 1 (1) point. In some embodiments, the clinical activity score (CAS) of the subject is reduced by 0 (0) point.

[0413] In some embodiments, a method is provided for treating thyroid-associated eye disease (TAO) in a subject or reducing its severity, wherein treatment using the antibody (i) reduces proptosis of one eyeball by at least 2 mm, (ii) does not entail worsening of the opposite eyeball (or opposite eye) by more than 2 mm, and (iii) reduces the CAS of the subject to one (1) or zero (0).

[0414] In some embodiments, a method for improving quality of life in a subject having thyroid-associated eye disease (TAO, also referred to as Graves' ophthalmopathy / Graves' orbital disease) is provided. In some embodiments, quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment or by either the visual function or appearance subscales thereof. In some embodiments, treatment causes an improvement of 8 points or more in the GO-QoL. In some embodiments, treatment causes an improvement in the function subscale of the GO-QoL. In some embodiments, treatment causes an improvement in the appearance subscale of the GO-QoL.

[0415] In some embodiments, a method is provided for treating diplopia or reducing its severity in subjects having thyroid-associated eye disease (TAO). In some embodiments, the diplopia is persistent diplopia. In some embodiments, the diplopia is non-persistent diplopia. In some embodiments, the diplopia is intermittent diplopia. In some embodiments, the improvement in diplopia or the reduction in its severity is maintained for at least 20 weeks after discontinuation of antibody administration. In some embodiments, the improvement in diplopia or the reduction in its severity is maintained for at least 50 weeks after discontinuation of antibody administration.

[0416] The severity of the disease may be measured in the following non-limiting embodiments. For example, in the case of lid aperture, the distance between the eyelid margins is measured (in mm) while the patient is sitting comfortably and looking straight ahead and at a distance. In the case of swelling of the eyelid, the measurement / evaluation is either 'absence / uncertain', 'moderate', or 'severe'. Redness of the eyelid is either absence or presence. Redness of the conjunctiva is either absence or presence. In some embodiments, conjunctival edema is either absence or presence. In some embodiments, inflammation of the sacrum or crescent fold is either absence or presence. Proptosis is measured in millimeters for each individual patient using the same Hertel exophthalmosometer and the same intercanthal distance. Subjective diplopia is scored from 0 to 3 (0 = no diplopia; 1 = intermittent, i.e., diplopia during visual inspection upon waking or when fatigued; 2 = non-persistent, i.e., diplopia during extreme visual inspection; 3 = persistent, i.e., continuous diplopia in a visual inspection or reading position). In the case of ocular muscle involvement, monocular movement is measured in degrees. Corneal involvement is either absent / punctate or keratopathy / ulcer. In the case of optic nerve involvement, i.e., best corrected visual acuity, color vision, optic disc, relative afferent pupillary defect, the condition is either absent or present. Additionally, if optic nerve compression is suspected, the visual field is examined. In some embodiments, the patient may be classified according to the following severity classifications. For example, visual-threatening thyroid eye disease: patients with dysthyroid optic neuropathy [DON] and / or corneal damage. This category justifies immediate intervention. Moderate to severe thyroid eye disease: Patients who do not have vision-threatening disease, but whose eye disease significantly impacts daily life to the extent that they are willing to accept the risk of immunosuppression (in active cases) or surgical intervention (in inactive cases).Patients with moderate to severe thyroid eye disease generally have any one or more of the following: eyelid retraction of 2 mm or more, moderate or severe soft tissue involvement, proptosis greater than 3 mm above the normal range for race and sex, and non-persistent or persistent diplopia. Mild thyroid eye disease: These are patients whose thyroid eye disease features have only a mild impact on daily life, insufficient to warrant immunosuppression or surgical treatment. They generally have only one or more of the following: mild eyelid retraction (less than 2 mm), mild soft tissue involvement, proptosis less than 3 mm above the normal range for race and sex, transient diplopia or absence of diplopia, and corneal exposure responsive to lubricants.

[0417] In some embodiments, the patient may be characterized by the Graves' Ophthalmopathy Quality of Life (GO-QoL) score. In addition to exophthalmos (or exophthalmos) and CAS, quality of life is also assessed using the GO Quality of Life (GO-QoL) questionnaire. This questionnaire is designed to determine improved quality of life after treatment using the method disclosed herein. In some embodiments, the questionnaire may determine reduced side effects or the absence of side effects after treatment with an antibody or its antigen-binding fragment according to the method disclosed herein compared to treatment with glucocorticoids. The GO-QoL is a 16-item self-administered questionnaire divided into two subsets and is used to assess the perceived impact of TED on the subject's (i) 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 (CB Terwee et al, 1998) is completed on Day 1 and at weeks 6, 12, and 24 (or PW) during the treatment period, and at months 7 and 12 (or PW) during the follow-up period. The GO-QoL is a 16-item self-administered questionnaire divided into two self-assessment subscales; one addresses the impact of visual function on daily activities, and the other assesses the impact on 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 ophthalmopathy has altered their appearance, caused others to react negatively toward them, caused social isolation, and caused them to attempt to cover their appearance. Each subscale consists of eight questions answered as Yes—Very much so, Yes—Somewhat so, or No—Not at all.Each question is scored from 0 to 2 points, and the total raw score is then mathematically converted to a 0 to 100 scale, where 0 indicates the most negative impact on quality of life and 100 indicates no impact. A change of 8 points or more on the 0 to 100 scale was found to be clinically significant. The combined score is obtained by taking the raw scores from both subscales and converting them back to a single 0 to 100 scale. The questionnaire has two self-assessment subscales. Each subscale has eight questions answered as (i) Yes—Very much so, (ii) Yes—Somewhat so, or (iii) No—Not at all. Each question is scored from 0 to 2 points, and the total raw score is then mathematically converted to a 0 to 100 scale, where 0 indicates the most negative impact on quality of life and 100 indicates no impact. A change of more than 8 points on a 0 to 100 scale is considered clinically significant. The combined score is obtained by taking the raw scores from both subscales and converting them back into a single 0 to 100 scale.

[0418] Patients may also be evaluated based on the presence or absence of the Gorman diplopia grade. The Gorman assessment of subjective diplopia includes four categories: absence of diplopia, diplopia when the patient is fatigued or immediately after waking up (intermittent), diplopia during extreme gaze (non-persistent), and continuous diplopia in a frontal or reading position (persistent). Patients are scored according to the grade of diplopia they are experiencing. An improvement of one grade or more is considered clinically significant.

[0419] In some embodiments, the method comprises administering an antibody such as that provided herein. In some embodiments, the antibody is administered as a first dose in an amount of about 1 mg / kg to about 5 mg / kg. In some embodiments, the antibody is administered as a first dose in an amount of about 5 mg / kg to about 10 mg / kg. In some embodiments, the antibody is administered as a first dose in an amount of about 10 mg / kg to about 20 mg / kg. In some embodiments, the antibody is administered as a subsequent dose in an amount of about 1 mg / kg to about 20 mg / kg. In some embodiments, the antibody is administered as a first dose in an amount of about 10 mg / kg antibody and as a subsequent dose in an amount of about 10 mg / kg antibody. In some embodiments, the antibody is administered as a first dose in an amount of about 3 mg / kg antibody and as a subsequent dose in an amount of about 3 mg / kg antibody. In some embodiments, the antibody is administered as a first dose in an amount of about 5 mg / kg antibody and as a subsequent dose in an amount of about 5 mg / kg antibody. In some embodiments, the antibody is administered as a first dose of about 20 mg / kg antibody and as a subsequent dose of about 20 mg / kg antibody. In some embodiments, the antibody is administered as a first dose of about 20 mg / kg antibody and as a subsequent dose of about 10 mg / kg antibody. In some embodiments, the subsequent dose is administered every 3 weeks for at least 12, 15, 18, or 21 weeks.

[0420] In some embodiments, the antibody is administered in a pharmaceutical composition such as that provided herein. In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of TAO. In some embodiments, 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.

[0421] In some embodiments, the method provided herein comprises administering to a subject an antibody or an antigen-binding fragment thereof that specifically binds to and inhibits IGF-IR. In some embodiments, the antibody is the same as that provided herein.

[0422] A kit useful for carrying out the embodiments described herein is also provided. The kit comprises a first container containing or associated with the antibody described above. The kit may also comprise another container containing or associated with a solution that is necessary or convenient for carrying out the embodiments. The container may be made of glass, plastic, or foil and may be a vial, bottle, pouch, tube, bag, etc. The kit may also include written information, such as a procedure for carrying out the embodiments, or analytical information, such as the amount of reagent contained in the first container means. The container may be in another container apparatus, e.g., a box or bag, along with the written information.

[0423] Another embodiment provided herein is a kit for detecting IGF-1R protein in biological samples. The kit comprises a container containing one or more antibodies that bind to an epitope of IGF-1R protein and instructions for using the antibodies to bind to IGF-1R protein to form an immunological complex and to detect the formation of the immunological complex, wherein the presence or absence of the immunological complex correlates with the presence or absence of IGF-1R protein in the sample. An example of a container includes a multi-well plate that enables simultaneous detection of IGF-1R protein in multiple samples.

[0424] In some embodiments, an antibody that binds to an IGF-1R protein is provided. In some embodiments, the antibody is isolated. In some embodiments, the antibody binds specifically. In some embodiments, the antibody binds to a properly folded IGF-1R protein. In some embodiments, the antibody is specific to a specific IGF-1R structural state (open or closed). In some embodiments, the antibody binds to an IGF-1R protein within a cell membrane. In some embodiments, the antibody binds to an IGF-1R protein within the cell membrane of an intact cell. In some embodiments, the antibody inhibits or neutralizes the function of the IGF-1R protein. As used herein, the term "neutralize" means that the activity or function of the protein is inhibited. Inhibition may be complete or partial. In some embodiments, the activity or function of the protein is inhibited by at least 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, or 99%. Percent inhibition may be based on the function or activity of the protein in the absence of the antibody. In some embodiments, the antibody inhibits glucose transport promoted by IGF-1R. In some embodiments, the antibody inhibits the internalization of the IGF-1R protein.

[0425] In some embodiments, the antibody comprises the sequence provided herein or the antigen-binding fragment thereof. In some embodiments, the antibody comprises the heavy chain CDR described herein or the antigen-binding fragment thereof. The heavy chain may be one or more of the heavy chains described herein. In some embodiments, the antibody comprises the light chain described herein or the antigen-binding fragment thereof.

[0426] In some embodiments, a method for treating, inhibiting, or improving an IGF-1R-associated pathology is provided. In some embodiments, the method comprises administering to a subject the antibody described herein or the pharmaceutical composition described herein to treat, inhibit, or improve an IGF-1R-associated pathology. In some embodiments, the pathology is as described herein.

[0427] In some embodiments, a method for detecting the presence or absence of IGF-1R in a sample is provided, the method comprises contacting the sample with one or more antibodies described herein and detecting binding to the IGF-1R antigen by the antibody. In some embodiments, detection of binding indicates the presence of the IGF-1R antigen, or the absence of detection of binding to the IGF-1R antigen indicates the absence of the IGF-1R antigen. Detection may 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 a protein under non-denaturing conditions. Non-denaturing conditions may be used so that the protein of interest may be detected in its native form or in a suitably folded form.

[0428] In some embodiments, a method for identifying a test antibody that binds to an epitope on an IGF-1R protein is provided, the method comprises 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 the sequence provided herein. In some embodiments, determining comprises mutating one or more residues of the epitope or protein and determining the binding of the test antibody to the mutated epitope, wherein if the mutation reduces the binding of the test antibody compared to a non-mutated epitope, the test antibody is deemed to bind to the corresponding epitope.

[0429] In some embodiments, a method for monitoring the internalization of IGF-1R from the cell surface is provided. In some embodiments, the method comprises contacting a cell with the anti-IGF-1R antibody provided herein and detecting the presence of IGF-1R within the cell or on the cell surface. Differences in cell surface expression may be measured, and internalization may be monitored and measured. This may be used to measure the effect of another molecule, such as a test agent, on regulating the internalization of the IGF-1R protein. Thus, the antibody provided herein may be used to identify a test agent that regulates (increases or decreases) the internalization of the IGF-1R protein. A test molecule that increases internalization, measured as a decrease in the binding of the anti-IGF-1R antibody to the IGF-1R protein on the cell surface, may be identified according to the method provided herein. A test molecule that decreases internalization, measured as an increase in the binding of the anti-IGF-1R antibody to the IGF-1R protein on the cell surface, may be identified according to the method provided herein. Surface expression can be measured by fluorescence, which can be achieved through a secondary antibody that recognizes the IGF-1R antibody or by labeling with the anti-IGF-1R antibody provided herein.

[0430] In some embodiments, a method for inhibiting IGF-1-stimulated receptor phosphorylation on cells is provided. In some embodiments, the method comprises contacting the cells with the antibody provided herein or a pharmaceutical composition containing the same. In some embodiments, contacting comprises administering the antibody or a pharmaceutical composition containing the same to a subject. In some embodiments, the cells are cells of the eye. In some embodiments, the subject has thyroid eye disease (TED) or is at risk thereof. In some embodiments, the antibody has an IC of about 0.2 nm, 0.15 nm, 0.10 nm, 0.09 nm or less. 50 It has. In some embodiments, IC 50IC50 is measured in an in vitro assay as provided herein or as exemplified in the Examples. In some embodiments, IC50 is measured in cells that are A549 cells or HOCF cells.

[0431] In some embodiments, a method for treating thyroid eye disease in a subject is provided, the method comprises administering to the subject an antibody provided herein or a pharmaceutical composition containing the same, wherein the antibody has a serum concentration in the subject of 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 after at least 1, 2, or 3 weeks after administration. In some embodiments, the serum concentration is measured after one, two, or three doses of the antibody or a pharmaceutical composition containing the same are administered to the subject.

[0432] A method is provided for inhibiting IGF-1-induced receptor autophosphorylation in a subject required in some embodiments by at least 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the method comprises administering the antibody provided herein or a pharmaceutical composition containing the same to a subject. In some embodiments, IGF-1-induced receptor autophosphorylation is inhibited in the eye or orbital region of the subject. In some embodiments, by inhibiting IGF-1-induced receptor autophosphorylation, the subject is treated for thyroid eye disease or the symptoms described herein are improved.

[0433] Enumerated embodiments

[0434] In some embodiments, the embodiments provided herein also include, but are not limited to, the following.

[0435] 1. An antibody comprising a variable heavy chain having an amino acid sequence at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence at least 90% identical to SEQ ID NO. 2,

[0436] The above variable heavy chain is

[0437] Glycine corresponding to position 26 of SEQ ID NO. 1, tyrosine corresponding to position 27 of SEQ ID NO. 1, threonine corresponding to position 28 of SEQ ID NO. 1, phenylalanine corresponding to position 29 of SEQ ID NO. 1, threonine corresponding to position 30 of SEQ ID NO. 1, serine corresponding to position 31 of SEQ ID NO. 1, tyrosine corresponding to position 32 of SEQ ID NO. 1, methionine corresponding to position 34 of SEQ ID NO. 1, isoleucine corresponding to position 51 of SEQ ID NO. 1, asparagine corresponding to position 59 of SEQ ID NO. 1, alanine corresponding to position 97 of SEQ ID NO. 1, arginine corresponding to position 98 of SEQ ID NO. 1, glycine corresponding to position 99 of SEQ ID NO. 1, proline corresponding to position 101 of SEQ ID NO. 1, aspartate corresponding to position 102 of SEQ ID NO. 1, tyrosine corresponding to position 103 of SEQ ID NO. 1, Lysine corresponding to position 108 of SEQ ID NO. 1, tyrosine corresponding to position 110 of SEQ ID NO. 1, phenylalanine corresponding to position 111 of SEQ ID NO. 1, aspartate corresponding to position 112 of SEQ ID NO. 1, or any combination thereof, arginine corresponding to position 40 of SEQ ID NO. 1, isoleucine corresponding to position 48 of SEQ ID NO. 1, lysine corresponding to position 67 of SEQ ID NO. 1, alanine corresponding to position 68 of SEQ ID NO. 1, leucine corresponding to position 70 of SEQ ID NO. 1, valine corresponding to position 72 of SEQ ID NO. 1, lysine corresponding to position 74 of SEQ ID NO. 1, alanine corresponding to position 79 of SEQ ID NO. 1, glutamine corresponding to position 82 of SEQ ID NO. 1, threonine corresponding to position 87 of SEQ ID NO. 1, serine corresponding to position 91 of SEQ ID NO. 1, and / or

[0438] A mutation corresponding to position 33 of SEQ ID NO. 1, e.g., W33H; a mutation corresponding to position 50 of SEQ ID NO. 1, e.g., E50H; a mutation corresponding to position 52 of SEQ ID NO. 1, e.g., N52H; a mutation corresponding to position 53 of SEQ ID NO. 1, e.g., P53H; a mutation corresponding to position 54 of SEQ ID NO. 1, e.g., S54H; a mutation corresponding to position 55 of SEQ ID NO. 1, e.g., N55H; a mutation corresponding to position 56 of SEQ ID NO. 1, e.g., G56H; a mutation corresponding to position 57 of SEQ ID NO. 1, e.g., R57H; a mutation corresponding to position 58 of SEQ ID NO. 1, e.g., T58H; a mutation corresponding to position 60 of SEQ ID NO. 1, e.g., Y60H; a mutation corresponding to position 61 of SEQ ID NO. 1, e.g., N61H; a mutation corresponding to position 62 of SEQ ID NO. 1, e.g., Q62H; SEQ ID NO. 1 A mutation corresponding to 63, e.g., K63H; a mutation corresponding to position 64 of SEQ ID NO. 1, e.g., F64H; a mutation corresponding to position 65 of SEQ ID NO. 1, e.g., Q65H; a mutation corresponding to position 66 of SEQ ID NO. 1, e.g., G66H; a mutation corresponding to position 96 of SEQ ID NO. 1, e.g., F96H; a mutation corresponding to position 100 of SEQ ID NO. 1, e.g., R100H; a mutation corresponding to position 104 of SEQ ID NO. 1, e.g., Y104H; a mutation corresponding to position 105 of SEQ ID NO. 1, e.g., G105H; a mutation corresponding to position 106 of SEQ ID NO. 1, e.g., S106H; a mutation corresponding to position 107 of SEQ ID NO. 1, e.g., S107H; a mutation corresponding to position 109 of SEQ ID NO. 1, e.g., W109H; a mutation corresponding to position 113 of SEQ ID NO. 1, e.g., V113H, a mutation corresponding to position 12 of SEQ ID NO. 1, e.g., V12K, a mutation corresponding to position 20 of SEQ ID NO. 1, e.g., L20V, a mutation corresponding to position 22 of SEQ ID NO. 1,For example, including C22A or C22K, a mutation corresponding to position 38 of SEQ ID NO. 1, e.g., K38R, a mutation corresponding to position 76 of SEQ ID NO. 1, e.g., S76T, or any combination thereof,

[0439] The above variable light chain is

[0440] Isoleucine corresponding to position 29 of SEQ ID NO. 2, valine corresponding to position 34 of SEQ ID NO. 2, glutamate corresponding to position 39 of SEQ ID NO. 2, asparagine corresponding to position 58 of SEQ ID NO. 2, glycine corresponding to position 96 of SEQ ID NO. 2, proline corresponding to position 100 of SEQ ID NO. 2, proline corresponding to position 101 of SEQ ID NO. 2, threonine corresponding to position 102 of SEQ ID NO. 2, threonine corresponding to position 7 of SEQ ID NO. 2, serine corresponding to position 14 of SEQ ID NO. 2, aspartate corresponding to position 17 of SEQ ID NO. 2, leucine corresponding to position 42 of SEQ ID NO. 2, lysine corresponding to position 44 of SEQ ID NO. 2, leucine corresponding to position 51 of SEQ ID NO. 2, alanine corresponding to position 72 of SEQ ID NO. 2, arginine corresponding to position 79 of SEQ ID NO. 2, or any combination thereof and / or

[0441] A mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, e.g., T36H; a mutation corresponding to position 38 of SEQ ID NO. 2, e.g., L38H; a mutation corresponding to position 55 of SEQ ID NO. 2, e.g., K55H; SEQ ID NO. 2, position An antibody comprising a mutation corresponding to 56, e.g., V56H; a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; a mutation corresponding to position 90 of SEQ ID NO. 2, e.g., I90V; or any combination thereof.

[0442] 2. In Embodiment 1, the antibody comprises a variable heavy chain having an amino acid sequence at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence at least 90% identical to SEQ ID NO. 2, wherein SEQ ID NO. 1 is a mutation at position 26, e.g., G26H; a mutation at position 27, e.g., Y27H; a mutation at position 28, e.g., T28H; a mutation at position 29, e.g., F29H; a mutation at position 30, e.g., T30H; a mutation at position 31, e.g., S31H; a mutation at position 32, e.g., Y32H; a mutation at position 34, e.g., M34H; a mutation at position 51, e.g., I51H; a mutation at position 59, e.g., N59H; a mutation at position 97, e.g., A97H; a mutation at position 98, e.g., R98H; a mutation at position 99, e.g., G99H; and at position 101 Mutation, e.g., P101H, mutation at position 102, e.g., D102H, mutation at position 103, e.g., Y103H, mutation at position 108, e.g., K108H, mutation at position 110, e.g., Y110H, mutation at position 111, e.g., F111H, mutation at position 112, e.g., D112H, mutation at position 40, e.g., R40A, mutation at position 48, e.g., I48M, mutation at position 67, e.g., K67R, mutation at position 68, e.g., A68V, mutation at position 70, e.g., L70M, mutation at position 72, e.g., V72R, mutation at position 74, e.g., K74T, mutation at position 79, e.g., A79V, mutation at position 82, e.g., Q82E, mutation at position 87, e.g., T87R, excluding a mutation at position 91, e.g., S91T, SEQ ID NO. 2 includes a mutation at position 29, e.g., I29H, a mutation at position 34, e.g.,Not including V34H, a mutation at position 37, e.g., Y37H, a mutation at position 39, e.g., E39H, a mutation at position 58, e.g., N58H, a mutation at position 96, e.g., G96H, a mutation at position 100, e.g., P100H, a mutation at position 101, e.g., P101H, a mutation at position 102, e.g., T102H, a mutation at position 7, e.g., T7S, a mutation at position 14, e.g., S14T, a mutation at position 17, e.g., D17Q, a mutation at position 42, e.g., L42Q, a mutation at position 44, e.g., K44R, a mutation at position 51, e.g., L51R, a mutation at position 72, e.g., A72S, a mutation at position 79, e.g., R79K, and

[0443] The above variable heavy chain is

[0444] A mutation corresponding to position 33 of SEQ ID NO. 1, e.g., W33H; a mutation corresponding to position 50 of SEQ ID NO. 1, e.g., E50H; a mutation corresponding to position 52 of SEQ ID NO. 1, e.g., N52H; a mutation corresponding to position 53 of SEQ ID NO. 1, e.g., P53H; a mutation corresponding to position 54 of SEQ ID NO. 1, e.g., S54H; a mutation corresponding to position 55 of SEQ ID NO. 1, e.g., N55H; a mutation corresponding to position 56 of SEQ ID NO. 1, e.g., G56H; a mutation corresponding to position 57 of SEQ ID NO. 1, e.g., R57H; a mutation corresponding to position 58 of SEQ ID NO. 1, e.g., T58H; a mutation corresponding to position 60 of SEQ ID NO. 1, e.g., Y60H; a mutation corresponding to position 61 of SEQ ID NO. 1, e.g., N61H; a mutation corresponding to position 62 of SEQ ID NO. 1, e.g., Q62H; SEQ ID NO. 1 A mutation corresponding to 63, e.g., K63H; a mutation corresponding to position 64 of SEQ ID NO. 1, e.g., F64H; a mutation corresponding to position 65 of SEQ ID NO. 1, e.g., Q65H; a mutation corresponding to position 66 of SEQ ID NO. 1, e.g., G66H; a mutation corresponding to position 96 of SEQ ID NO. 1, e.g., F96H; a mutation corresponding to position 100 of SEQ ID NO. 1, e.g., R100H; a mutation corresponding to position 104 of SEQ ID NO. 1, e.g., Y104H; a mutation corresponding to position 105 of SEQ ID NO. 1, e.g., G105H; a mutation corresponding to position 106 of SEQ ID NO. 1, e.g., S106H; a mutation corresponding to position 107 of SEQ ID NO. 1, e.g., S107H; a mutation corresponding to position 109 of SEQ ID NO. 1, e.g., W109H; a mutation corresponding to position 113 of SEQ ID NO. 1, e.g., V113H, a mutation corresponding to position 12 of SEQ ID NO. 1, e.g., V12K, a mutation corresponding to position 20 of SEQ ID NO. 1, e.g., L20V, a mutation corresponding to position 22 of SEQ ID NO. 1,For example, including C22A or C22K, a mutation corresponding to position 38 of SEQ ID NO. 1, e.g., K38R, a mutation corresponding to position 76 of SEQ ID NO. 1, e.g., S76T, or any combination thereof,

[0445] The above variable light chain is

[0446] A mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, e.g., T36H; a mutation corresponding to position 38 of SEQ ID NO. 2, e.g., L38H; a mutation corresponding to position 55 of SEQ ID NO. 2, e.g., K55H; SEQ ID NO. 2, position An antibody comprising a mutation corresponding to 56, e.g., V56H; a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; a mutation corresponding to position 90 of SEQ ID NO. 2, e.g., I90V; or any combination thereof.

[0447] 3. In any one of embodiments 1 to 2, the variable heavy chain is an antibody comprising a W33H mutation.

[0448] 4. In embodiment 3, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 10, an antibody.

[0449] 5. In any one of embodiments 1 to 4, the variable heavy chain is an antibody comprising an E50H mutation.

[0450] 6. In embodiment 5, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 12, an antibody.

[0451] 7. In any one of embodiments 1 to 6, the variable heavy chain comprises an N52H mutation, an antibody.

[0452] 8. In embodiment 7, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 14, an antibody.

[0453] 9. An antibody in any one of embodiments 1 to 8, wherein the variable heavy chain comprises a P53H mutation.

[0454] 10. An antibody in embodiment 9, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 15.

[0455] 11. An antibody in any one of embodiments 1 to 10, wherein the variable heavy chain comprises an S54H mutation.

[0456] 12. An antibody according to embodiment 11, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 16.

[0457] 13. An antibody in any one of embodiments 1 to 12, wherein the variable heavy chain comprises an N55H mutation.

[0458] 14. An antibody according to embodiment 13, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 17.

[0459] 15. An antibody in any one of embodiments 1 to 14, wherein the variable heavy chain comprises a G56H mutation.

[0460] 16. An antibody according to embodiment 15, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 18.

[0461] 17. An antibody in any one of embodiments 1 to 16, wherein the variable heavy chain comprises an R57H mutation.

[0462] 18. An antibody according to embodiment 17, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 19.

[0463] 19. An antibody in any one of embodiments 1 to 18, wherein the variable heavy chain comprises a T58H mutation.

[0464] 20. An antibody according to embodiment 19, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 20.

[0465] 21. An antibody in any one of embodiments 1 to 20, wherein the variable heavy chain comprises a Y60H mutation.

[0466] 22. An antibody according to embodiment 21, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 22.

[0467] 23. An antibody in any one of embodiments 1 to 22, wherein the variable heavy chain comprises an N61H mutation.

[0468] 24. An antibody according to embodiment 23, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 23.

[0469] 25. An antibody in any one of embodiments 1 to 24, wherein the variable heavy chain comprises a Q62H mutation.

[0470] 26. An antibody according to embodiment 25, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 24.

[0471] 27. An antibody in any one of embodiments 1 to 26, wherein the variable heavy chain comprises a K63H mutation.

[0472] 28. An antibody according to embodiment 27, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 25.

[0473] 29. An antibody in any one of embodiments 1 to 28, wherein the variable heavy chain comprises an F64H mutation.

[0474] 30. An antibody according to embodiment 29, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 26.

[0475] 31. An antibody in any one of embodiments 1 to 30, wherein the variable heavy chain comprises a Q65H mutation.

[0476] 32. An antibody according to embodiment 31, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 27.

[0477] 33. An antibody in any one of embodiments 1 to 32, wherein the variable heavy chain comprises a G66H mutation.

[0478] 34. An antibody according to embodiment 33, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 28.

[0479] 35. An antibody in any one of embodiments 1 to 34, wherein the variable heavy chain comprises an F96H mutation.

[0480] 36. An antibody according to embodiment 35, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 29.

[0481] 37. An antibody in any one of embodiments 1 to 36, wherein the variable heavy chain comprises an R100H mutation.

[0482] 38. An antibody according to embodiment 37, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 33.

[0483] 39. An antibody in any one of embodiments 1 to 38, wherein the variable heavy chain comprises a Y104H mutation.

[0484] 40. An antibody according to embodiment 39, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 37.

[0485] 41. An antibody in any one of embodiments 1 to 40, wherein the variable heavy chain comprises a G105H mutation.

[0486] 42. An antibody according to embodiment 41, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 38.

[0487] 43. An antibody in any one of embodiments 1 to 42, wherein the variable heavy chain comprises an S106H mutation.

[0488] 44. An antibody according to embodiment 43, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 39.

[0489] 45. An antibody in any one of embodiments 1 to 44, wherein the variable heavy chain comprises an S107H mutation.

[0490] 46. ​​An antibody according to embodiment 45, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 40.

[0491] 47. An antibody in any one of embodiments 1 to 46, wherein the variable heavy chain comprises a W109H mutation.

[0492] 48. An antibody according to embodiment 47, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 42.

[0493] 49. An antibody in any one of embodiments 1 to 48, wherein the variable heavy chain comprises a V113H mutation.

[0494] 50. An antibody according to embodiment 49, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 46.

[0495] 51. An antibody in any one of embodiments 1 to 50, wherein the variable heavy chain comprises a V12K mutation.

[0496] 52. An antibody according to embodiment 51, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 77.

[0497] 53. An antibody in any one of embodiments 1 to 52, wherein the variable heavy chain comprises an L20V mutation.

[0498] 54. An antibody according to embodiment 53, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 78.

[0499] 55. An antibody in any one of embodiments 1 to 54, wherein the variable heavy chain comprises a C22A or C22K mutation.

[0500] 56. An antibody according to embodiment 55, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 79 or 80.

[0501] 57. An antibody in any one of embodiments 1 to 56, wherein the variable heavy chain comprises a K38R mutation.

[0502] 58. An antibody according to embodiment 57, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 81.

[0503] 59. An antibody in any one of embodiments 1 to 58, wherein the variable heavy chain comprises an S76T mutation.

[0504] 60. An antibody according to embodiment 59, wherein the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 89.

[0505] 61. An antibody in any one of embodiments 1 to 60, wherein the variable light chain comprises an R24H mutation.

[0506] 62. An antibody according to embodiment 61, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 47.

[0507] 63. An antibody in any one of embodiments 1 to 62, wherein the variable light chain comprises an S25H mutation.

[0508] 64. An antibody according to embodiment 63, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 48.

[0509] 65. An antibody in any one of embodiments 1 to 64, wherein the variable light chain comprises an S26H mutation.

[0510] 66. An antibody according to embodiment 65, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 49.

[0511] 67. An antibody in any one of embodiments 1 to 66, wherein the variable light chain comprises a Q27H mutation.

[0512] 68. An antibody according to embodiment 67, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 50.

[0513] 69. An antibody in any one of embodiments 1 to 68, wherein the variable light chain comprises an S28H mutation.

[0514] 70. An antibody according to embodiment 69, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 51.

[0515] 71. An antibody in any one of embodiments 1 to 70, wherein the variable light chain comprises a V30H mutation.

[0516] 72. An antibody according to embodiment 71, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 53.

[0517] 73. An antibody in any one of embodiments 1 to 72, wherein the variable light chain comprises an S32H mutation.

[0518] 74. An antibody according to embodiment 73, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 54.

[0519] 75. An antibody in any one of embodiments 1 to 74, wherein the variable light chain comprises an N33H mutation.

[0520] 76. An antibody according to embodiment 75, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 55.

[0521] 77. An antibody in any one of embodiments 1 to 76, wherein the variable light chain comprises an N35H mutation.

[0522] 78. An antibody according to embodiment 77, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 57.

[0523] 79. An antibody in any one of embodiments 1 to 78, wherein the variable light chain comprises a T36H mutation.

[0524] 80. An antibody according to embodiment 79, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 58.

[0525] 81. An antibody in any one of embodiments 1 to 80, wherein the variable light chain comprises an L38H mutation.

[0526] 82. An antibody according to embodiment 81, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 60.

[0527] 83. An antibody in any one of embodiments 1 to 82, wherein the variable light chain comprises a K55H mutation.

[0528] 84. An antibody according to embodiment 83, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 62.

[0529] 85. An antibody in any one of embodiments 1 to 84, wherein the variable light chain comprises a V56H mutation.

[0530] 86. An antibody according to embodiment 85, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 63.

[0531] 87. An antibody in any one of embodiments 1 to 86, wherein the variable light chain comprises an S57H mutation.

[0532] 88. An antibody according to embodiment 87, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 64.

[0533] 89. An antibody in any one of embodiments 1 to 88, wherein the variable light chain comprises an R59H mutation.

[0534] 90. An antibody according to embodiment 89, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 66.

[0535] 91. An antibody in any one of embodiments 1 to 90, wherein the variable light chain comprises an F60H mutation.

[0536] 92. An antibody according to embodiment 91, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 67.

[0537] 93. An antibody in any one of embodiments 1 to 92, wherein the variable light chain comprises an S61H mutation.

[0538] 94. An antibody according to embodiment 93, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 68.

[0539] 95. An antibody in any one of embodiments 1 to 94, wherein the variable light chain comprises an F94H mutation.

[0540] 96. An antibody according to embodiment 95, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 69.

[0541] 97. An antibody in any one of embodiments 1 to 96, wherein the variable light chain comprises a Q95H mutation.

[0542] 98. An antibody according to embodiment 97, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 70.

[0543] 99. An antibody in any one of embodiments 1 to 98, wherein the variable light chain comprises an S97H mutation.

[0544] 100. An antibody according to embodiment 99, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 72.

[0545] 101. An antibody in any one of embodiments 1 to 100, wherein the variable light chain comprises a V99H mutation.

[0546] 102. An antibody according to embodiment 101, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 73.

[0547] 103. An antibody in any one of embodiments 1 to 102, wherein the variable light chain comprises a Y41F mutation.

[0548] 104. An antibody according to embodiment 103, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 97.

[0549] 105. An antibody in any one of embodiments 1 to 104, wherein the variable light chain comprises an L88V mutation.

[0550] 106. An antibody according to embodiment 105, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 103.

[0551] 107. An antibody in any one of embodiments 1 to 106, wherein the variable light chain comprises an I90V mutation.

[0552] 108. An antibody according to embodiment 107, wherein the variable light chain comprises the amino acid sequence of SEQ ID NO. 104.

[0553] 109. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 10.

[0554] 110. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 12.

[0555] 111. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 14.

[0556] 112. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 15.

[0557] 113. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 16.

[0558] 114. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 17.

[0559] 115. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 18.

[0560] 116. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 19.

[0561] 117. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 20.

[0562] 118. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 22.

[0563] 119. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 23.

[0564] 120. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 24.

[0565] 121. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 25.

[0566] 122. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 26.

[0567] 123. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 27.

[0568] 124. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 28.

[0569] 125. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 29.

[0570] 126. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 33.

[0571] 127. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 37.

[0572] 128. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 38.

[0573] 129. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 39.

[0574] 130. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 40.

[0575] 131. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 42.

[0576] 132. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 46.

[0577] 133. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 77.

[0578] 134. An antibody in embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 78.

[0579] 135. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 79.

[0580] 136. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 80.

[0581] 137. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 81.

[0582] 138. An antibody according to embodiment 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO. 89.

[0583] 139. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 47.

[0584] 140. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 48.

[0585] 141. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 49.

[0586] 142. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 50.

[0587] 143. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 51.

[0588] 144. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 53.

[0589] 145. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 54.

[0590] 146. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 55.

[0591] 147. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 57.

[0592] 148. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 58.

[0593] 149. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 60.

[0594] 150. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 62.

[0595] 151. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 63.

[0596] 152. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 64.

[0597] 153. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 66.

[0598] 154. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 67.

[0599] 155. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 68.

[0600] 156. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 69.

[0601] 157. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 70.

[0602] 158. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 72.

[0603] 159. An antibody according to embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 73.

[0604] 160. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 97.

[0605] 161. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 103.

[0606] 162. An antibody in embodiment 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO. 104.

[0607] 163. A method for treating thyroid-associated eye disease (TAO) or its symptoms or reducing its severity, comprising administering an antibody of any one of embodiments 1 to 162 to a subject.

[0608] 164. A method for reducing proptosis in the eye of a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0609] 165. A method for treating thyroid eye disease in a subject, comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0610] 166. A method for reducing the clinical activity score (CAS) of thyroid-associated eye disease (TAO) in a subject, comprising administering an antibody of any one of embodiments 1 to 162 to a subject.

[0611] 167. A method for a) reducing proptosis by at least 2 mm and b) reducing the Clinical Activity Score (CAS) in a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0612] 168. A method in any one of embodiments 163 to 167, wherein the protrusion of the eyeball is reduced by at least 2 mm.

[0613] 169. A method in any one of embodiments 163 to 167, wherein the protrusion of the eyeball is reduced by at least 3 mm.

[0614] 170. A method in any one of embodiments 163 to 167, wherein the protrusion of the eyeball is reduced by at least 4 mm.

[0615] 171. A method in any one of embodiments 163 to 167, wherein the clinical activity score (CAS) of the subject is reduced by at least 2 points.

[0616] 172. A method in which, in any one of embodiments 163 to 167, the clinical activity score (CAS) of the subject is reduced to one (1) point.

[0617] 173. A method in any one of embodiments 163 to 167, wherein the clinical activity score (CAS) of the subject is reduced to zero (0) points.

[0618] 174. A method for treating thyroid-associated eye disease (TAO) in a subject or reducing the severity thereof, comprising administering an antibody of any one of embodiments 1 to 162 to the subject, wherein the treatment using said antibody (i) reduces proptosis of one eyeball by at least 2 mm, (ii) does not entail worsening of the opposite eyeball (or opposite eye) by more than 2 mm, and (iii) reduces said CAS of said subject to one (1) or zero (0).

[0619] 175. A method for improving the quality of life in a subject having thyroid-associated eye disease (TAO, also referred to as Graves' ophthalmopathy / Graves' orbital disease), comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0620] 176. A method according to embodiment 175, wherein the quality of life is measured by the Graves' ophthalmopathy Quality of Life (GO-QoL) assessment, or by any one of the visual function or appearance subscales thereof.

[0621] 177. A method according to embodiment 176, wherein the treatment causes an improvement of 8 points or more in the GO-QoL.

[0622] 178. In embodiment 176, the treatment causes improvement in the functional subscale of the GO-QoL.

[0623] 179. In embodiment 176, the treatment causes improvement in the appearance subscale of the GO-QoL.

[0624] 180. A method for treating diplopia or reducing the severity thereof in a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0625] 181. Method according to embodiment 180, wherein the diplopia is persistent diplopia.

[0626] 182. Method according to embodiment 180, wherein the diplopia is non-persistent diplopia.

[0627] 183. Method according to embodiment 180, wherein the diplopia is intermittent diplopia.

[0628] 184. A method according to embodiment 180, wherein the improvement of diplopia or the reduction in the severity thereof is maintained for at least 20 weeks after discontinuation of antibody administration.

[0629] 185. A method according to embodiment 180, wherein the improvement of diplopia or the reduction in the severity thereof is maintained for at least 50 weeks after discontinuation of antibody administration.

[0630] 186. A method in any one of embodiments 163 to 185, wherein the antibody is administered as a first dose at a dosage of about 1 mg / kg to about 5 mg / kg of antibody.

[0631] 187. A method in any one of embodiments 163 to 185, wherein the antibody is administered as a first dose at a dosage of about 5 mg / kg to about 10 mg / kg of antibody.

[0632] 188. A method in any one of embodiments 163 to 185, wherein the antibody is subsequently administered at a dose of about 5 mg / kg to about 20 mg / kg of antibody.

[0633] 189. A method in any one of embodiments 163 to 185, wherein the antibody is administered in an amount of about 10 mg / kg antibody as a first dose and about 20 mg / kg antibody as a subsequent dose.

[0634] 190. A method according to embodiment 189, wherein the subsequent dose is administered every 3 weeks for at least 21 weeks.

[0635] 191. A method in any one of embodiments 163 to 190, wherein the antibody or its antigen-binding fragment is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a diabody, a single-chain antibody, a multi-specific antibody, Fab, Fab', F(ab')2, Fv, or scFv.

[0636] 192. A method in any one of embodiments 163 to 191, wherein the antibody or its antigen-binding fragment is additionally administered into a pharmaceutical composition comprising a pharmaceutically acceptable diluent or excipient or carrier.

[0637] 193. A method according to embodiment 192, wherein the pharmaceutical composition further comprises one or more pharmaceutically active compounds for the treatment of the TAO.

[0638] 194. Method according to embodiment 192 or 193, 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.

[0639] 195. A method in any one of embodiments 163 to 194, wherein the antibody or its antigen-binding fragment is administered directly to the eye, the anterior chamber of the eye, the vitreous cavity of the eye, the supracoracial space, or the posterior orbital venous sinus.

[0640] 196. A method according to embodiment 195, wherein the antibody or its antigen-binding fragment is administered via injection.

[0641] 197. Method according to embodiment 196, wherein the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a supracorachaelinal injection, or an anterior chamber injection.

[0642] 198. A method for increasing the internalization of IGF-1R in a cell, comprising contacting the cell with an antibody of any one of embodiments 1 to 162.

[0643] 199. A method according to embodiment 198, wherein the contacting comprises administering an antibody of any one of embodiments 1 to 162 to a subject.

[0644] 200. In embodiment 174, the subject has thyroid eye disease (TED) or is at risk thereof.

[0645] 201. A method for inhibiting IGF-1-stimulated receptor phosphorylation on a cell, comprising contacting the cell with an antibody of any one of embodiments 1 to 162.

[0646] 202. A method according to embodiment 201, wherein the contacting comprises administering an antibody of any one of embodiments 1 to 162 to a subject.

[0647] 203. The method of embodiment 202, wherein the subject has thyroid eye disease (TED) or is at risk thereof.

[0648] 204. A method in any one of embodiments 201 to 203, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, or 0.09 nm or less.

[0649] 205. In embodiment 204, the IC50 is measured in an in vitro test such as the test provided herein.

[0650] 206. A method in any one of embodiments 201 to 205, wherein the cell is an A549 cell or a HOCF cell.

[0651] 207. A method for treating thyroid eye disease in a subject, comprising administering an antibody of any one of embodiments 1 to 162, wherein the antibody has a serum concentration in the subject of 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 after at least 1, 2, or 3 weeks of administration.

[0652] 208. A method according to embodiment 207, wherein the antibody is administered intravenously.

[0653] 209. A method in any one of embodiments 207 to 208, wherein the antibody is administered at a dose of about 20 mg / kg.

[0654] 210. A method in any one of embodiments 207 to 208, wherein the antibody is administered at least or about once a week, once every two weeks, once every three weeks, or once every four weeks.

[0655] 211. A method for inhibiting IGF-1-induced receptor autophosphorylation in cells by at least 95%, 96%, 97%, 98%, 99%, or 100%, comprising contacting the cells with an antibody of any one of Examples 1 to 162.

[0656] 212. A method according to embodiment 211, wherein the inhibition of the IGF-1-induced receptor autophosphorylation is measured in comparison to receptor autophosphorylation induced in the absence of the antibody.

[0657] 213. A method according to embodiment 211 or 212, wherein the contacting comprises administering the antibody containing the above to a subject.

[0658] 214. The method of embodiment 213, wherein the subject has thyroid eye disease (TED) or is at risk thereof.

[0659] 215. A method for inhibiting IGF-1-induced receptor autophosphorylation in a subject requiring at least 95%, 96%, 97%, 98%, or 99% or 100%, comprising administering an antibody of any one of embodiments 1 to 162 to the subject.

[0660] 216. Method according to embodiment 190, wherein the subject has thyroid eye disease (TED) or is at risk thereof.

[0661] 217. A method in which, in either embodiment 215 or 216, the antibody is administered intravenously.

[0662] The subject matter is now described with reference to the following examples. These examples are provided solely for illustrative purposes, and the claims should not be interpreted as being limited to these examples, but rather as encompassing any and all variations that become apparent as a result of the teachings provided herein. A person skilled in the art will readily recognize various non-critical parameters that may be changed or modified to yield substantially similar results.

[0663] Examples

[0664] Example 1: A variant of the anti-IGF-1R antibody VRDN-01100 binds to IGF-1R

[0665] Mutation screening of variants of the anti-IGF-1R antibody VRDN-01100 was performed by analyzing the kinetics and affinity of antibody variants containing single-point mutations in variable heavy and light chains. HD293 cells were transiently transformed to express the antibody variants. The supernatant was collected and analyzed via biolayer interference to characterize the ability of the antibody variants to bind to IGF-1R by comparison with VRDN-01100. Antibody variants were immobilized on an anti-human IgG Fc capture biosensor, and biolayer interference was performed using 100 nM recombinant human IGF-1R protein as analytes for the association and dissociation steps. The assay was performed at a temperature of 37 °C and a shaking speed of 1000 rpm. For each antibody variant, the association rate (K a ), equilibrium dissociation constant (K D ) and dissociation rate constant (k d Not only ) but also k d The multiple change was determined. The multiple change is calculated as the ratio of the variant dissociation rate constant to the VRDN-01100 dissociation rate constant; accordingly, a multiple change value of <1 indicates improved binding to IGF-1R by the antibody variant compared to VRDN-01100, and a multiple change value of >1 indicates reduced binding. The following Table 4As shown in [figure], a specific mutation (e.g., VRDN-01204) improved binding affinity to IGF-1R, while another mutation (e.g., VRDN-01205) decreased binding affinity.

[0666]

[0667]

[0668]

[0669]

[0670]

[0671]

[0672]

[0673]

[0674] These data demonstrate that these variants can bind to IGF-1R with a specific affinity and can be used to treat conditions such as thyroid eye disease.

[0675] The disclosures of each and all patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety. Although various embodiments have been disclosed by reference to specific aspects, it is evident that other aspects and variations of these embodiments may be devised by a person skilled in the art without departing from the true spirit and scope of the embodiments. The appended claims are intended to be interpreted as including all such aspects and equivalent variations.

Claims

Claim 1 An antibody comprising a variable heavy chain having an amino acid sequence at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence at least 90% identical to SEQ ID NO. 2, wherein the variable heavy chain comprises glycine corresponding to position 26 of SEQ ID NO. 1, tyrosine corresponding to position 27 of SEQ ID NO. 1, threonine corresponding to position 28 of SEQ ID NO. 1, phenylalanine corresponding to position 29 of SEQ ID NO. 1, threonine corresponding to position 30 of SEQ ID NO. 1, serine corresponding to position 31 of SEQ ID NO. 1, tyrosine corresponding to position 32 of SEQ ID NO. 1, methionine corresponding to position 34 of SEQ ID NO. 1, isoleucine corresponding to position 51 of SEQ ID NO. 1, asparagine corresponding to position 59 of SEQ ID NO. 1, alanine corresponding to position 97 of SEQ ID NO. 1, arginine corresponding to position 98 of SEQ ID NO. 1, and glycine corresponding to position 99 of SEQ ID NO.

1. Proline corresponding to position 101 of SEQ ID NO. 1, aspartate corresponding to position 102 of SEQ ID NO. 1, tyrosine corresponding to position 103 of SEQ ID NO. 1, lysine corresponding to position 108 of SEQ ID NO. 1, tyrosine corresponding to position 110 of SEQ ID NO. 1, phenylalanine corresponding to position 111 of SEQ ID NO. 1, aspartate corresponding to position 112 of SEQ ID NO. 1, arginine corresponding to position 40 of SEQ ID NO. 1, isoleucine corresponding to position 48 of SEQ ID NO. 1, lysine corresponding to position 67 of SEQ ID NO. 1, alanine corresponding to position 68 of SEQ ID NO. 1, leucine corresponding to position 70 of SEQ ID NO. 1, valine corresponding to position 72 of SEQ ID NO. 1, lysine corresponding to position 74 of SEQ ID NO. 1, alanine corresponding to position 79 of SEQ ID NO. 1, glutamine corresponding to position 82 of SEQ ID NO. 1, Threonine corresponding to position 87 of SEQ ID NO. 1, serine corresponding to position 91 of SEQ ID NO. 1, or any combination thereof and / or a mutation corresponding to position 33 of SEQ ID NO. 1, e.g., W33H, a mutation corresponding to position 50 of SEQ ID NO. 1, e.g., E50H,A mutation corresponding to position 52 of SEQ ID NO. 1, e.g., N52H; a mutation corresponding to position 53 of SEQ ID NO. 1, e.g., P53H; a mutation corresponding to position 54 of SEQ ID NO. 1, e.g., S54H; a mutation corresponding to position 55 of SEQ ID NO. 1, e.g., N55H; a mutation corresponding to position 56 of SEQ ID NO. 1, e.g., G56H; a mutation corresponding to position 57 of SEQ ID NO. 1, e.g., R57H; a mutation corresponding to position 58 of SEQ ID NO. 1, e.g., T58H; a mutation corresponding to position 60 of SEQ ID NO. 1, e.g., Y60H; a mutation corresponding to position 61 of SEQ ID NO. 1, e.g., N61H; a mutation corresponding to position 62 of SEQ ID NO. 1, e.g., Q62H; a mutation corresponding to position 63 of SEQ ID NO. 1, e.g., K63H; a mutation corresponding to position 64 of SEQ ID NO. 1, e.g., F64H; SEQ ID NO. 1, A mutation corresponding to 65, e.g., Q65H; a mutation corresponding to position 66 of SEQ ID NO. 1, e.g., G66H; a mutation corresponding to position 96 of SEQ ID NO. 1, e.g., F96H; a mutation corresponding to position 100 of SEQ ID NO. 1, e.g., R100H; a mutation corresponding to position 104 of SEQ ID NO. 1, e.g., Y104H; a mutation corresponding to position 105 of SEQ ID NO. 1, e.g., G105H; a mutation corresponding to position 106 of SEQ ID NO. 1, e.g., S106H; a mutation corresponding to position 107 of SEQ ID NO. 1, e.g., S107H; a mutation corresponding to position 109 of SEQ ID NO. 1, e.g., W109H; a mutation corresponding to position 113 of SEQ ID NO. 1, e.g., V113H; a mutation corresponding to position 12 of SEQ ID NO. 1, e.g., V12K; a mutation corresponding to position 20 of SEQ ID NO. 1, e.g., L20V, a mutation corresponding to position 22 of SEQ ID NO. 1, e.g., C22A or C22K, a mutation corresponding to position 38 of SEQ ID NO. 1, e.g., K38R,A mutation corresponding to position 76 of SEQ ID NO. 1, e.g., S76T, or any combination thereof, and / or the variable light chain comprises isoleucine corresponding to position 29 of SEQ ID NO. 2, valine corresponding to position 34 of SEQ ID NO. 2, glutamate corresponding to position 39 of SEQ ID NO. 2, asparagine corresponding to position 58 of SEQ ID NO. 2, glycine corresponding to position 96 of SEQ ID NO. 2, proline corresponding to position 100 of SEQ ID NO. 2, proline corresponding to position 101 of SEQ ID NO. 2, threonine corresponding to position 102 of SEQ ID NO. 2, threonine corresponding to position 7 of SEQ ID NO. 2, serine corresponding to position 14 of SEQ ID NO. 2, aspartate corresponding to position 17 of SEQ ID NO. 2, leucine corresponding to position 42 of SEQ ID NO. 2, lysine corresponding to position 44 of SEQ ID NO. 2, leucine corresponding to position 51 of SEQ ID NO. 2 Alanine corresponding to position 72 of 2, arginine corresponding to position 79 of SEQ ID NO. 2, or any combination thereof and / or a mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, e.g., T36H; a mutation corresponding to position 38 of SEQ ID NO. 2 For example, L38H, a mutation corresponding to position 55 of SEQ ID NO. 2, for example, K55H, a mutation corresponding to position 56 of SEQ ID NO. 2, for example, V56H,An antibody comprising a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; a mutation corresponding to position 90 of SEQ ID NO. 2, e.g., I90V; or any combination thereof. Claim 2 In claim 1, the antibody comprises a variable heavy chain having an amino acid sequence at least 90% identical to SEQ ID NO. 1 and a variable light chain having an amino acid sequence at least 90% identical to SEQ ID NO. 2, wherein SEQ ID NO. 1 is a mutation at position 26, e.g., G26H; a mutation at position 27, e.g., Y27H; a mutation at position 28, e.g., T28H; a mutation at position 29, e.g., F29H; a mutation at position 30, e.g., T30H; a mutation at position 31, e.g., S31H; a mutation at position 32, e.g., Y32H; a mutation at position 34, e.g., M34H; a mutation at position 51, e.g., I51H; a mutation at position 59, e.g., N59H; a mutation at position 97, e.g., A97H; a mutation at position 98, e.g., R98H; a mutation at position 99, e.g., G99H; a mutation at position 101, e.g., P101H, mutation at position 102, e.g., D102H, mutation at position 103, e.g., Y103H, mutation at position 108, e.g., K108H, mutation at position 110, e.g., Y110H, mutation at position 111, e.g., F111H, mutation at position 112, e.g., D112H, mutation at position 40, e.g., R40A, mutation at position 48, e.g., I48M, mutation at position 67, e.g., K67R, mutation at position 68, e.g., A68V, mutation at position 70, e.g., L70M, mutation at position 72, e.g., V72R, mutation at position 74, e.g., K74T, mutation at position 79, e.g., A79V, mutation at position 82, e.g., Q82E, mutation at position 87, e.g., T87R, position Not including a mutation at 91, e.g., S91T, the above SEQ ID NO. 2 includes a mutation at position 29, e.g., I29H, a mutation at position 34, e.g.,V34H, a mutation at position 37, e.g., Y37H, a mutation at position 39, e.g., E39H, a mutation at position 58, e.g., N58H, a mutation at position 96, e.g., G96H, a mutation at position 100, e.g., P100H, a mutation at position 101, e.g., P101H, a mutation at position 102, e.g., T102H, a mutation at position 7, e.g., T7S, a mutation at position 14, e.g., S14T, a mutation at position 17, e.g., D17Q, a mutation at position 42, e.g., L42Q, a mutation at position 44, e.g., K44R, a mutation at position 51, e.g., L51R, a mutation at position 72, e.g., A72S, a mutation at position 79, e.g., R79K, are not included, and the variable heavy chain does not include a mutation corresponding to position 33 of SEQ ID NO. 1, e.g., W33H, a mutation corresponding to position 50 of SEQ ID NO. 1, e.g., E50H, a mutation corresponding to position 52 of SEQ ID NO. 1, e.g., N52H, a mutation corresponding to position 53 of SEQ ID NO. 1, e.g., P53H, a mutation corresponding to position 54 of SEQ ID NO. 1, e.g., S54H, a mutation corresponding to position 55 of SEQ ID NO. 1, e.g., N55H, a mutation corresponding to position 56 of SEQ ID NO. 1, e.g., G56H, a mutation corresponding to position 57 of SEQ ID NO. 1, e.g., R57H, a mutation corresponding to position 58 of SEQ ID NO. 1, e.g., T58H, a mutation corresponding to position 60 of SEQ ID NO. 1, e.g., Y60H, a mutation corresponding to position 61 of SEQ ID NO. 1, e.g., N61H, a mutation corresponding to position 62 of SEQ ID NO. 1, e.g., Q62H, a mutation corresponding to position 63 of SEQ ID NO. 1, e.g., K63H, A mutation corresponding to position 64 of SEQ ID NO. 1, e.g., F64H; a mutation corresponding to position 65 of SEQ ID NO. 1, e.g., Q65H; a mutation corresponding to position 66 of SEQ ID NO. 1,For example, G66H, a mutation corresponding to position 96 of SEQ ID NO. 1, for example, F96H, a mutation corresponding to position 100 of SEQ ID NO. 1, for example, R100H, a mutation corresponding to position 104 of SEQ ID NO. 1, for example, Y104H, a mutation corresponding to position 105 of SEQ ID NO. 1, for example, G105H, a mutation corresponding to position 106 of SEQ ID NO. 1, for example, S106H, a mutation corresponding to position 107 of SEQ ID NO. 1, for example, S107H, a mutation corresponding to position 109 of SEQ ID NO. 1, for example, W109H, a mutation corresponding to position 113 of SEQ ID NO. 1, for example, V113H, a mutation corresponding to position 12 of SEQ ID NO. 1, for example, V12K, a mutation corresponding to position 20 of SEQ ID NO. 1, for example, L20V, a mutation corresponding to position 22 of SEQ ID NO. 1, for example, C22A or C22K, A mutation corresponding to position 38 of SEQ ID NO. 1, e.g., K38R; a mutation corresponding to position 76 of SEQ ID NO. 1, e.g., S76T; or any combination thereof; and / or the variable light chain comprises a mutation corresponding to position 24 of SEQ ID NO. 2, e.g., R24H; a mutation corresponding to position 25 of SEQ ID NO. 2, e.g., S25H; a mutation corresponding to position 26 of SEQ ID NO. 2, e.g., S26H; a mutation corresponding to position 27 of SEQ ID NO. 2, e.g., Q27H; a mutation corresponding to position 28 of SEQ ID NO. 2, e.g., S28H; a mutation corresponding to position 30 of SEQ ID NO. 2, e.g., V30H; a mutation corresponding to position 32 of SEQ ID NO. 2, e.g., S32H; a mutation corresponding to position 33 of SEQ ID NO. 2, e.g., N33H; a mutation corresponding to position 35 of SEQ ID NO. 2, e.g., N35H; a mutation corresponding to position 36 of SEQ ID NO. 2, For example, T36H, a mutation corresponding to position 38 of SEQ ID NO. 2, for example, L38H, a mutation corresponding to position 55 of SEQ ID NO. 2, for example, K55H,A mutation corresponding to position 56 of SEQ ID NO. 2, e.g., V56H; a mutation corresponding to position 57 of SEQ ID NO. 2, e.g., S57H; a mutation corresponding to position 59 of SEQ ID NO. 2, e.g., R59H; a mutation corresponding to position 60 of SEQ ID NO. 2, e.g., F60H; a mutation corresponding to position 61 of SEQ ID NO. 2, e.g., S61H; a mutation corresponding to position 94 of SEQ ID NO. 2, e.g., F94H; a mutation corresponding to position 95 of SEQ ID NO. 2, e.g., Q95H; a mutation corresponding to position 97 of SEQ ID NO. 2, e.g., S97H; a mutation corresponding to position 99 of SEQ ID NO. 2, e.g., V99H; a mutation corresponding to position 41 of SEQ ID NO. 2, e.g., Y41F; a mutation corresponding to position 88 of SEQ ID NO. 2, e.g., L88V; a mutation corresponding to position 90 of SEQ ID NO. 2, e.g., I90V; or any combination thereof, comprising Antibody., Claim 3 An antibody according to any one of claims 1 to 2, wherein the variable heavy chain comprises a W33H mutation. Claim 4 In paragraph 3, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 10, an antibody. Claim 5 An antibody according to any one of claims 1 to 4, wherein the variable heavy chain comprises an E50H mutation. Claim 6 In claim 5, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 12, an antibody. Claim 7 An antibody according to any one of claims 1 to 6, wherein the variable heavy chain comprises an N52H mutation. Claim 8 In claim 7, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 14, an antibody. Claim 9 In any one of claims 1 to 8, the variable heavy chain is an antibody comprising a P53H mutation. Claim 10 In claim 9, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 15, an antibody. Claim 11 An antibody according to any one of claims 1 to 10, wherein the variable heavy chain comprises an S54H mutation. Claim 12 In claim 11, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 16, an antibody. Claim 13 An antibody according to any one of claims 1 to 12, wherein the variable heavy chain comprises an N55H mutation. Claim 14 In claim 13, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 17, an antibody. Claim 15 An antibody according to any one of claims 1 to 14, wherein the variable heavy chain comprises a G56H mutation. Claim 16 In claim 15, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 18, an antibody. Claim 17 An antibody according to any one of claims 1 to 16, wherein the variable heavy chain comprises an R57H mutation. Claim 18 In claim 17, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 19, an antibody. Claim 19 An antibody according to any one of claims 1 to 18, wherein the variable heavy chain comprises a T58H mutation. Claim 20 In claim 19, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 20, an antibody. Claim 21 An antibody according to any one of claims 1 to 20, wherein the variable heavy chain comprises a Y60H mutation. Claim 22 In claim 21, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 22, an antibody. Claim 23 An antibody according to any one of claims 1 to 22, wherein the variable heavy chain comprises an N61H mutation. Claim 24 In paragraph 23, the above variable heavy chain is an antibody comprising the amino acid sequence of SEQ ID NO.

23. Claim 25 An antibody according to any one of claims 1 to 24, wherein the variable heavy chain comprises a Q62H mutation. Claim 26 In claim 25, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 24, an antibody. Claim 27 An antibody according to any one of claims 1 to 26, wherein the variable heavy chain comprises a K63H mutation. Claim 28 In claim 27, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 25, an antibody. Claim 29 In any one of claims 1 to 28, the variable heavy chain is an antibody comprising an F64H mutation. Claim 30 In claim 29, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 26, an antibody. Claim 31 An antibody according to any one of claims 1 to 30, wherein the variable heavy chain comprises a Q65H mutation. Claim 32 In claim 31, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 27, an antibody. Claim 33 An antibody according to any one of claims 1 to 32, wherein the variable heavy chain comprises a G66H mutation. Claim 34 In paragraph 33, the above variable heavy chain is an antibody comprising the amino acid sequence of SEQ ID NO.

28. Claim 35 An antibody according to any one of claims 1 to 34, wherein the variable heavy chain comprises an F96H mutation. Claim 36 In claim 35, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 29, an antibody. Claim 37 An antibody according to any one of claims 1 to 36, wherein the variable heavy chain comprises an R100H mutation. Claim 38 In claim 37, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 33, an antibody. Claim 39 An antibody according to any one of claims 1 to 38, wherein the variable heavy chain comprises a Y104H mutation. Claim 40 In claim 39, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 37, an antibody. Claim 41 An antibody according to any one of claims 1 to 40, wherein the variable heavy chain comprises a G105H mutation. Claim 42 In claim 41, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 38, an antibody. Claim 43 An antibody according to any one of claims 1 to 42, wherein the variable heavy chain comprises an S106H mutation. Claim 44 In paragraph 43, the above variable heavy chain is an antibody comprising the amino acid sequence of SEQ ID NO.

39. Claim 45 An antibody according to any one of claims 1 to 44, wherein the variable heavy chain comprises an S107H mutation. Claim 46 In claim 45, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 40, an antibody. Claim 47 An antibody according to any one of claims 1 to 46, wherein the variable heavy chain comprises a W109H mutation. Claim 48 In claim 47, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 42, an antibody. Claim 49 An antibody according to any one of claims 1 to 48, wherein the variable heavy chain comprises a V113H mutation. Claim 50 In claim 49, the above variable heavy chain is an antibody comprising the amino acid sequence of SEQ ID NO.

46. Claim 51 An antibody according to any one of claims 1 to 50, wherein the variable heavy chain comprises a V12K mutation. Claim 52 In claim 51, the variable heavy chain comprises the amino acid sequence of SEQ ID NO. 77, an antibody. Claim 53 In any one of claims 1 to 52, the variable heavy chain is an antibody comprising an L20V mutation. Claim 54 In paragraph 53, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 78, an antibody. Claim 55 An antibody according to any one of claims 1 to 54, wherein the variable heavy chain comprises a C22A or C22K mutation. Claim 56 In claim 55, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 79 or 80, an antibody. Claim 57 An antibody according to any one of claims 1 to 56, wherein the variable heavy chain comprises a K38R mutation. Claim 58 In claim 57, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 81, an antibody. Claim 59 An antibody according to any one of claims 1 to 58, wherein the variable heavy chain comprises an S76T mutation. Claim 60 In claim 59, the above variable heavy chain comprises the amino acid sequence of SEQ ID NO. 89, an antibody. Claim 61 An antibody according to any one of claims 1 to 60, wherein the variable light chain comprises an R24H mutation. Claim 62 In claim 61, the variable light chain comprises the amino acid sequence of SEQ ID NO. 47, an antibody. Claim 63 An antibody according to any one of claims 1 to 62, wherein the variable light chain comprises an S25H mutation. Claim 64 In paragraph 63, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 48, an antibody. Claim 65 An antibody according to any one of claims 1 to 64, wherein the variable light chain comprises an S26H mutation. Claim 66 In claim 65, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 49, an antibody. Claim 67 An antibody according to any one of claims 1 to 66, wherein the variable light chain comprises a Q27H mutation. Claim 68 In claim 67, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 50, an antibody. Claim 69 An antibody according to any one of claims 1 to 68, wherein the variable light chain comprises an S28H mutation. Claim 70 In claim 69, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 51, an antibody. Claim 71 In any one of claims 1 to 70, the variable light chain is an antibody comprising a V30H mutation. Claim 72 In claim 71, the variable light chain comprises the amino acid sequence of SEQ ID NO. 53, an antibody. Claim 73 An antibody according to any one of claims 1 to 72, wherein the variable light chain comprises an S32H mutation. Claim 74 In paragraph 73, the variable light chain comprises the amino acid sequence of SEQ ID NO. 54, an antibody. Claim 75 An antibody according to any one of claims 1 to 74, wherein the variable light chain comprises an N33H mutation. Claim 76 In claim 75, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 55, an antibody. Claim 77 An antibody according to any one of claims 1 to 76, wherein the variable light chain comprises an N35H mutation. Claim 78 In claim 77, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 57, an antibody. Claim 79 An antibody according to any one of claims 1 to 78, wherein the variable light chain comprises a T36H mutation. Claim 80 In claim 79, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 58, an antibody. Claim 81 An antibody according to any one of claims 1 to 80, wherein the variable light chain comprises an L38H mutation. Claim 82 In claim 81, the variable light chain comprises the amino acid sequence of SEQ ID NO. 60, an antibody. Claim 83 An antibody according to any one of claims 1 to 82, wherein the variable light chain comprises a K55H mutation. Claim 84 In paragraph 83, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 62, an antibody. Claim 85 In any one of claims 1 to 84, the variable light chain is an antibody comprising a V56H mutation. Claim 86 In claim 85, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 63, an antibody. Claim 87 An antibody according to any one of claims 1 to 86, wherein the variable light chain comprises an S57H mutation. Claim 88 In claim 87, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 64, an antibody. Claim 89 An antibody according to any one of claims 1 to 88, wherein the variable light chain comprises an R59H mutation. Claim 90 In claim 89, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 66, an antibody. Claim 91 An antibody according to any one of claims 1 to 90, wherein the variable light chain comprises an F60H mutation. Claim 92 In claim 91, the variable light chain comprises the amino acid sequence of SEQ ID NO. 67, an antibody. Claim 93 An antibody according to any one of claims 1 to 92, wherein the variable light chain comprises an S61H mutation. Claim 94 In paragraph 93, the variable light chain comprises the amino acid sequence of SEQ ID NO. 68, an antibody. Claim 95 An antibody according to any one of claims 1 to 94, wherein the variable light chain comprises an F94H mutation. Claim 96 In claim 95, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 69, an antibody. Claim 97 An antibody according to any one of claims 1 to 96, wherein the variable light chain comprises a Q95H mutation. Claim 98 In claim 97, the above variable light chain comprises the amino acid sequence of SEQ ID NO. 70, an antibody. Claim 99 An antibody according to any one of claims 1 to 98, wherein the variable light chain comprises an S97H mutation. Claim 100 In claim 99, the variable light chain comprises the amino acid sequence of SEQ ID NO. 72, an antibody. Claim 101 An antibody according to any one of claims 1 to 100, wherein the variable light chain comprises a V99H mutation. Claim 102 In claim 101, the variable light chain comprises the amino acid sequence of SEQ ID NO. 73, an antibody. Claim 103 An antibody according to any one of claims 1 to 102, wherein the variable light chain comprises a Y41F mutation. Claim 104 In claim 103, the variable light chain comprises the amino acid sequence of SEQ ID NO. 97, an antibody. Claim 105 In any one of claims 1 to 104, the variable light chain is an antibody comprising an L88V mutation. Claim 106 In claim 105, the variable light chain comprises the amino acid sequence of SEQ ID NO. 103, an antibody. Claim 107 In any one of claims 1 to 106, the variable light chain is an antibody comprising an I90V mutation. Claim 108 In claim 107, the variable light chain comprises the amino acid sequence of SEQ ID NO. 104, an antibody. Claim 109 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

10. Claim 110 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

12. Claim 111 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

14. Claim 112 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

15. Claim 113 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

16. Claim 114 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

17. Claim 115 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

18. Claim 116 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

19. Claim 117 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

20. Claim 118 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

22. Claim 119 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

23. Claim 120 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

24. Claim 121 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

25. Claim 122 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

26. Claim 123 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

27. Claim 124 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

28. Claim 125 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

29. Claim 126 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

33. Claim 127 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

37. Claim 128 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

38. Claim 129 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

39. Claim 130 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

40. Claim 131 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

42. Claim 132 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

46. Claim 133 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

77. Claim 134 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

78. Claim 135 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

79. Claim 136 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

80. Claim 137 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

81. Claim 138 An antibody according to claim 1 or 2, wherein the variable light chain has the amino acid sequence of SEQ ID NO. 2 and the variable heavy chain has the amino acid sequence of SEQ ID NO.

89. Claim 139 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

47. Claim 140 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

48. Claim 141 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

49. Claim 142 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

50. Claim 143 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

51. Claim 144 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

53. Claim 145 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

54. Claim 146 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

55. Claim 147 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

57. Claim 148 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

58. Claim 149 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

60. Claim 150 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

62. Claim 151 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

63. Claim 152 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

64. Claim 153 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

66. Claim 154 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

67. Claim 155 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

68. Claim 156 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

69. Claim 157 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

70. Claim 158 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

72. Claim 159 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

73. Claim 160 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

97. Claim 161 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

103. Claim 162 An antibody according to claim 1 or 2, wherein the variable heavy chain has the amino acid sequence of SEQ ID NO. 1 and the variable light chain has the amino acid sequence of SEQ ID NO.

104. Claim 163 A method for treating thyroid-associated eye disease (TAO) or its symptoms or reducing its severity, comprising administering an antibody of any one of claims 1 to 162 to a subject. Claim 164 A method for reducing proptosis in the eye of a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 165 A method for treating thyroid eye disease in a subject, comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 166 A method for reducing the clinical activity score (CAS) of thyroid-associated eye disease (TAO) in a subject, comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 167 A method for a) reducing proptosis by at least 2 mm and b) reducing the Clinical Activity Score (CAS) in a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 168 A method according to any one of claims 163 to 167, wherein the protrusion of the eyeball is reduced by at least 2 mm. Claim 169 A method according to any one of claims 163 to 167, wherein the protrusion of the eyeball is reduced by at least 3 mm. Claim 170 A method according to any one of claims 163 to 167, wherein the protrusion of the eyeball is reduced by at least 4 mm. Claim 171 A method according to any one of claims 163 to 167, wherein the clinical activity score (CAS) of the subject is reduced by at least 2 points. Claim 172 A method in which, in any one of claims 163 to 167, the clinical activity score (CAS) of the subject is reduced to one (1) point. Claim 173 A method in which, in any one of claims 163 to 167, the clinical activity score (CAS) of the subject is reduced to zero (0). Claim 174 A method for treating thyroid-associated eye disease (TAO) in a subject or reducing its severity, comprising administering an antibody of any one of claims 1 to 162 to the subject, wherein the treatment using said antibody (i) reduces proptosis of one eyeball by at least 2 mm, (ii) does not entail worsening of the opposite eyeball (or opposite eye) by more than 2 mm, and (iii) reduces said CAS of said subject to one (1) or zero (0). Claim 175 A method for improving the quality of life in a subject having thyroid-associated eye disease (TAO, also referred to as Graves' ophthalmopathy / Graves' orbital disease), comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 176 In paragraph 175, the quality of life is measured by the Graves' Ophthalmopathy Quality of Life (GO-QoL) assessment, or by either of its visual function or appearance subscales. Claim 177 In paragraph 176, the above treatment is a method that causes an improvement of 8 points or more in the GO-QoL. Claim 178 In paragraph 176, the above treatment is a method that causes improvement in the above functional subscale of the GO-QoL. Claim 179 In paragraph 176, the above treatment is a method that causes improvement in the appearance subscale of the above GO-QoL. Claim 180 A method for treating diplopia or reducing the severity thereof in a subject having thyroid-associated eye disease (TAO), comprising administering an antibody of any one of claims 1 to 162 to the subject. Claim 181 In paragraph 180, the above diplopia is persistent diplopia, method. Claim 182 In paragraph 180, the above diplopia is a non-persistent diplopia, method. Claim 183 In paragraph 180, the above diplopia is intermittent diplopia. Claim 184 A method according to claim 180, wherein the improvement of diplopia or reduction in its severity is maintained for at least 20 weeks after discontinuation of antibody administration. Claim 185 A method according to claim 180, wherein the improvement of diplopia or reduction in its severity is maintained for at least 50 weeks after discontinuation of antibody administration. Claim 186 A method according to any one of claims 163 to 185, wherein the antibody is administered as a first dose at an antibody dosage of about 1 mg / kg to about 5 mg / kg. Claim 187 A method according to any one of claims 163 to 185, wherein the antibody is administered as a first dose at an antibody dosage of about 5 mg / kg to about 10 mg / kg. Claim 188 A method according to any one of claims 163 to 185, wherein the antibody is subsequently administered at a dose of about 5 mg / kg to about 20 mg / kg of antibody. Claim 189 A method according to any one of claims 163 to 185, wherein the antibody is administered in an amount of about 10 mg / kg antibody as a first dose and about 20 mg / kg antibody as a subsequent dose. Claim 190 In paragraph 189, the method wherein the subsequent dose is administered every 3 weeks for at least 21 weeks. Claim 191 A method according to any one of claims 163 to 190, wherein the antibody or its antigen-binding fragment is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, a diabody, a single-chain antibody, a multi-specific antibody, Fab, Fab', F(ab')2, Fv, or scFv. Claim 192 A method according to any one of claims 163 to 191, wherein the antibody or its antigen-binding fragment is additionally administered in a pharmaceutical composition comprising a pharmaceutically acceptable diluent or excipient or carrier. Claim 193 In claim 192, the method wherein the pharmaceutical composition further comprises one or more pharmaceutical active compounds for the treatment of the TAO. Claim 194 The method of claim 192 or 193, 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. Claim 195 A method according to any one of claims 163 to 194, wherein the antibody or its antigen-binding fragment is administered directly to the eye, the anterior part of the eye, the vitreous cavity of the eye, the supracorbital space, or the posterior orbital sinus. Claim 196 In paragraph 195, the method wherein the antibody or its antigen-binding fragment is administered via injection. Claim 197 In paragraph 196, the injection is a method in which the injection is an intravitreal injection, an intraorbital injection, a retroorbital injection, a supracoradioscopic injection, or an anterior injection. Claim 198 A method for increasing the internalization of IGF-1R in a cell, comprising contacting the cell with an antibody of any one of claims 1 to 162. Claim 199 A method according to claim 198, wherein the above contact comprises administering an antibody of any one of claims 1 to 162 to a subject. Claim 200 In paragraph 174, the subject has thyroid eye disease (TED) or is at risk thereof. Claim 201 A method for inhibiting IGF-1-stimulated receptor phosphorylation on a cell, comprising contacting the cell with an antibody of any one of claims 1 to 162. Claim 202 A method according to claim 201, wherein the contacting comprises administering an antibody of any one of claims 1 to 162 to a subject. Claim 203 In paragraph 202, the subject has thyroid eye disease (TED) or is at risk thereof. Claim 204 A method according to any one of claims 201 to 203, wherein the antibody has an IC50 of about 0.2 nm, 0.15 nm, 0.10 nm, or 0.09 nm or less. Claim 205 In paragraph 204, the above IC50 is a method measured in an in vitro test such as the test provided herein. Claim 206 A method according to any one of claims 201 to 205, wherein the cell is an A549 cell or a HOCF cell. Claim 207 A method for treating thyroid eye disease in a subject, comprising administering an antibody of any one of claims 1 to 162, wherein the antibody has a serum concentration in the subject of 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 after at least 1, 2, or 3 weeks of administration. Claim 208 In paragraph 207, the above antibody is administered intravenously. Claim 209 A method according to any one of claims 207 to 208, wherein the antibody is administered at a dose of about 20 mg / kg. Claim 210 A method according to any one of claims 207 to 208, wherein the antibody is administered at least or about once a week, once every two weeks, once every three weeks, or once every four weeks. Claim 211 A method for inhibiting IGF-1-induced receptor autophosphorylation in cells by at least 95%, 96%, 97%, 98%, or 99% or 100%, comprising contacting said cells with an antibody of any one of claims 1 to 162. Claim 212 A method according to claim 211, wherein the inhibition of the IGF-1-induced receptor autophosphorylation is measured in comparison to receptor autophosphorylation induced in the absence of the antibody. Claim 213 A method according to claim 211 or 212, wherein the contacting comprises administering the antibody containing the above to a subject. Claim 214 In paragraph 213, the above subject has thyroid eye disease (TED) or is at risk thereof. Claim 215 A method for inhibiting IGF-1-induced receptor autophosphorylation in a subject requiring at least 95%, 96%, 97%, 98%, or 99% or 100%, comprising administering an antibody of any one of claims 1 to 162 to said subject. Claim 216 In paragraph 190, the method, wherein the subject has thyroid eye disease (TED) or is at risk thereof. Claim 217 A method according to either claim 215 or 216, wherein the antibody is administered intravenously.

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