Antibodies that bind interleukin 4 receptor alpha and methods of use

Abstract

Described herein are novel and improved antibodies that bind to interleukin 4 [alpha] receptor (IL-4R [alpha]) and methods of use thereof. In certain aspects, methods of inhibiting the biological activity of IL-4R alpha are described herein. In certain aspects, pharmaceutical compositions comprising the anti-IL-4R [alpha] antibody are described herein. In certain aspects, the antibodies and methods described herein are used to treat inflammatory diseases or disorders associated with elevated levels of IL-4 and / or IgE.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to co-pending U.S. Provisional Patent Application No. 63 / 446,763, filed February 17, 2023; U.S. Provisional Patent Application No. 63 / 462,864, filed April 28, 2023; U.S. Provisional Patent Application No. 63 / 596,501, filed November 6, 2023; and U.S. Provisional Patent Application No. 63 / 596,504, filed November 6, 2023, each of which is incorporated herein by reference in its entirety. Technical Field

[0003] Described herein are novel and improved antibodies that bind to the interleukin 4α receptor (IL-4Rα) and methods of use thereof, eg, for treating inflammatory diseases or disorders associated with elevated levels of IL-4 and / or IgE. Background Art

[0004] Interleukin 4 (IL-4) and interleukin 13 (IL-13) share a common receptor component, the interleukin-4 receptor alpha (IL-4Rα) chain (Nelms, K. et al. (1999) Annu. Rev. Immunol. 17:701-738) and (Jensen, PL (2000) Stem Cells 18:61-62). IL-4Rα pairs with a common γc chain to form a type I IL-4R complex found primarily in hematopoietic cells and exclusive for IL-4. IL-4Rα also pairs with the interleukin-13 receptor alpha 1 (IL-13Rα1) subunit to form a type II IL-4R that binds both IL-4 and IL-13. Type II receptors are expressed on both hematopoietic and non-hematopoietic cells. IL-4 (also known as B cell stimulating factor or BSF-1) was originally characterized by its ability to stimulate B cell proliferation in response to low concentrations of antibodies against surface immunoglobulins. IL-4 has been shown to have a wide range of biological activities, including growth stimulation of T cells, mast cells, granulocytes, megakaryocytes, and erythrocytes. IL-4 induces the expression of class II major histocompatibility complex molecules in resting B cells and enhances the secretion of IgE and IgG1 isotypes by stimulated B cells.

[0005] The biological and immune functions of B lymphocytes, monocytes, dendritic cells, and fibroblasts are all affected by IL-4 and IL-13. These cytokines interact with IL-4R to initiate the type 2 inflammatory pathway, which causes Th2 cell differentiation, inflammation, and mucus production. The type 2 inflammatory pathway is first activated in allergic diseases by abnormal cytokine release caused by an imbalance between Th1 and Th2 differentiation. Activated Th2 cells release cytokines, including IL-4, IL-13, and IL-31, which prompt downstream B cells to undergo changes and produce IgE antibodies. Mast cells and basophils are then required to degranulate and release inflammatory substances. At the same time, the secreted IL-4 and IL-13 continue to bind to their corresponding receptors, such as IL-4Rα, which repeatedly promote TH2 differentiation and subsequent inflammation.

[0006] The important role of IL-4 and IL-13 in the type 2 inflammatory pathway identifies IL-4Rα as a potential target for individuals who exhibit an aberrant type 2 inflammatory response. Unlike drug therapies that target only IL-4 and IL-13, targeting IL-4Rα would target both IL-4 and IL-13. Therefore, there is a need for novel antibodies that bind to IL-4Rα that can modulate type 2 inflammation to treat immune disorders. Summary of the Invention

[0007] In a first aspect, disclosed herein are antibodies, e.g., recombinant human antibodies, that specifically bind to human interleukin-4 receptor alpha (hIL-4Rα). In certain embodiments, the antibodies are characterized by the ability to bind to hIL-4Rα with high affinity and neutralize hIL-4Rα activity. In specific embodiments, the human antibodies are capable of blocking the binding of the hIL-13 / hIL-13R1 complex to hIL-4Rα and thereby inhibiting hIL-13 signaling. The antibodies may be full-length antibodies (e.g., IgG1 or IgG4 antibodies) or may comprise only an antigen-binding portion (e.g., Fab, F(ab')2, or scFv fragments) and may be modified to achieve functionality, e.g., to eliminate residual effector function (Reddy et al. (2000) J. Immunol. 164: 1925-1933) and / or to extend half-life.

[0008] In certain aspects, the present invention provides an isolated antibody that binds to interleukin (IL)-4 receptor alpha (IL-4Rα), the antibody comprising a) a variable heavy (VH) chain sequence having three heavy chain CDR sequences: CDR-H1, CDR-H2, and CDR-H3; and b) a variable light (VL) chain sequence having three light chain CDR sequences: CDR-L1, CDR-L2, and CDR-L3; wherein: a. CDR-H1 comprises a sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 1-4, 66-70, and 187-191; b. CDR-H2 comprises a sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 5-16 and 71-90; c. CDR-H3 comprises a sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 17-25 and 92-99; d. CDR-L1 comprises a sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 26-40 and 100-107; e. CDR-L2 comprises a sequence selected from the group consisting of sequences set forth in SEQ ID NOs: ID NOs: 41-52 and 108-112; and f. CDR-L3 comprises a sequence selected from the group consisting of SEQ ID NOs: 53-65.

[0009] In some embodiments, the antibody comprises: a. CDR-H1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 1-4; b. CDR-H2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 5-16; c. CDR-H3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 17-25; d. CDR-L1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 26-40; e. CDR-L2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 41-52; and f. CDR-L3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 53-65.

[0010] In some embodiments, the antibody comprises: a. CDR-H1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 66-70; b. CDR-H2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 71-78; c. CDR-H3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 17-25; d. CDR-L1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 26-40; e. CDR-L2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 41-52; and f. CDR-L3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 53-65.

[0011] In some embodiments, the antibody comprises: a. CDR-H1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 187-191; b. CDR-H2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 79-90; c. CDR-H3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 92-99; d. CDR-L1, which comprises a sequence selected from the sequence shown in SEQ ID NO: 100-107; e. CDR-L2, which comprises a sequence selected from the sequence shown in SEQ ID NO: 108-112; and f. CDR-L3, which comprises a sequence selected from the sequence shown in SEQ ID NO: 53-65.

[0012] In some embodiments, the antibody does not comprise:

[0013] a. CDR-H1 as shown in SEQ ID NO: 1; CDR-H2 as shown in SEQ ID NO: 5; CDR-H3 as shown in SEQ ID NO: 17; CDR-L1 as shown in SEQ ID NO: 26; CDR-L2 as shown in SEQ ID NO: 41; and CDR-L3 as shown in SEQ ID NO: 53; or

[0014] b. CDR-H1 as shown in SEQ ID NO: 67; CDR-H2 as shown in SEQ ID NO: 71; CDR-H3 as shown in SEQ ID NO: 17; CDR-L1 as shown in SEQ ID NO: 26; CDR-L2 as shown in SEQ ID NO: 41; and CDR-L3 as shown in SEQ ID NO: 53; or

[0015] c. CDR-H1 shown in SEQ ID NO: 187; CDR-H2 shown in SEQ ID NO: 79; CDR-H3 shown in SEQ ID NO: 91; CDR-L1 shown in SEQ ID NO: 100; CDR-L2 shown in SEQ ID NO: 108; and CDR-L3 shown in SEQ ID NO: 53.

[0016] In some embodiments, the antibody does not comprise any combination of: a. CDR-H1 shown in any one of SEQ ID NO: 1, 67 or 187; b. CDR-H2 shown in any one of SEQ ID NO: 5, 71 or 79; c. CDR-H3 shown in any one of SEQ ID NO: 17 or 91; d. CDR-L1 shown in any one of SEQ ID NO: 26 or 100; e. CDR-L2 shown in any one of SEQ ID NO: 41 or 108; and f. CDR-L3 shown in SEQ ID NO: 53.

[0017] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 54.

[0018] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 62, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0019] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0020] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0021] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 56.

[0022] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0023] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 46 or 109; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 58.

[0024] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 59.

[0025] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 68, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0026] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0027] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0028] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0029] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0030] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0031] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 21 or 96; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0032] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 62.

[0033] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0034] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 23, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0035] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 56.

[0036] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 23 or 97; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0037] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20, 18, or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0038] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0039] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 36 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0040] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 37 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0041] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 12, 74, or 86; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0042] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 13, 71, or 87; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0043] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0044] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0045] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 66, or 187; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 38 or 105; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 48 or 111; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 63.

[0046] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 41 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 64.

[0047] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 49 or 112; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 63.

[0048] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0049] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 58.

[0050] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 50 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0051] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0052] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 67, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 67, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 40 or 107; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 52 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0053] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0054] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22, 18, or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0055] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 65.

[0056] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 62.

[0057] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0058] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0059] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0060] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0061] In some embodiments, the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 25 or 99; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 53.

[0062] In some embodiments, the antibody comprises a heavy chain variable domain (VH) sequence selected from the group consisting of sequences shown in SEQ ID NOs: 113-145.

[0063] In some embodiments, the antibody comprises a light chain variable domain (VL) sequence selected from the group consisting of the sequences shown in SEQ ID NOs: 146-186.

[0064] In some embodiments, the antibody comprises a VH sequence selected from the group consisting of sequences shown in SEQ ID NOs: 113-145, and a VL sequence selected from the group consisting of sequences shown in SEQ ID NOs: 146-186.

[0065] In certain aspects, described herein is an isolated antibody that binds IL-4Rα, wherein the antibody comprises a VH sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 113-145 and a VL sequence selected from the group consisting of sequences set forth in SEQ ID NOs: 146-186.

[0066] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:147.

[0067] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:148.

[0068] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:115 and the VL sequence shown in SEQ ID NO:149.

[0069] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:150.

[0070] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:116 and the VL sequence shown in SEQ ID NO:151.

[0071] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:117 and the VL sequence shown in SEQ ID NO:152.

[0072] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:118 and the VL sequence shown in SEQ ID NO:153.

[0073] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:117 and the VL sequence shown in SEQ ID NO:154.

[0074] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:119 and the VL sequence shown in SEQ ID NO:155.

[0075] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:120 and the VL sequence shown in SEQ ID NO:156.

[0076] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:120 and the VL sequence shown in SEQ ID NO:157.

[0077] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:158.

[0078] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:121 and the VL sequence shown in SEQ ID NO:158.

[0079] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:159.

[0080] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:123 and the VL sequence shown in SEQ ID NO:159.

[0081] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:124 and the VL sequence shown in SEQ ID NO:160.

[0082] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:125 and the VL sequence shown in SEQ ID NO:161.

[0083] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:126 and the VL sequence shown in SEQ ID NO:162.

[0084] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:127 and the VL sequence shown in SEQ ID NO:163.

[0085] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:128 and the VL sequence shown in SEQ ID NO:164.

[0086] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:124 and the VL sequence shown in SEQ ID NO:158.

[0087] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:129 and the VL sequence shown in SEQ ID NO:165.

[0088] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:130 and the VL sequence shown in SEQ ID NO:166.

[0089] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:131 and the VL sequence shown in SEQ ID NO:167.

[0090] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:132 and the VL sequence shown in SEQ ID NO:159.

[0091] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:133 and the VL sequence shown in SEQ ID NO:159.

[0092] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:121 and the VL sequence shown in SEQ ID NO:168.

[0093] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:134 and the VL sequence shown in SEQ ID NO:169.

[0094] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:113 and the VL sequence shown in SEQ ID NO:170.

[0095] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:134 and the VL sequence shown in SEQ ID NO:171.

[0096] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:135 and the VL sequence shown in SEQ ID NO:172.

[0097] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:136 and the VL sequence shown in SEQ ID NO:173.

[0098] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:131 and the VL sequence shown in SEQ ID NO:174.

[0099] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:137 and the VL sequence shown in SEQ ID NO:175.

[0100] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:176.

[0101] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:138 and the VL sequence shown in SEQ ID NO:177.

[0102] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:134 and the VL sequence shown in SEQ ID NO:178.

[0103] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:139 and the VL sequence shown in SEQ ID NO:179.

[0104] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:140 and the VL sequence shown in SEQ ID NO:180.

[0105] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:141 and the VL sequence shown in SEQ ID NO:181.

[0106] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:142 and the VL sequence shown in SEQ ID NO:182.

[0107] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:143 and the VL sequence shown in SEQ ID NO:183.

[0108] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:114 and the VL sequence shown in SEQ ID NO:184.

[0109] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:144 and the VL sequence shown in SEQ ID NO:185.

[0110] In some embodiments, the antibody comprises the VH sequence shown in SEQ ID NO:145 and the VL sequence shown in SEQ ID NO:186.

[0111] In some embodiments, the antibody is a humanized, fully human, or chimeric antibody.

[0112] In some embodiments, the antibody is a fully human antibody.

[0113] In some embodiments, the antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM.

[0114] In some embodiments, the human Fc region comprises a human heavy chain constant region of class IgG and a subclass selected from IgGl, IgG2, IgG3, and IgG4.

[0115] In some embodiments, the human Fc region comprises a human IgG1 Fc.

[0116] In some embodiments, the human Fc region comprises a human IgG4 Fc.

[0117] In some embodiments, the human Fc region comprises a human IgG2 Fc.

[0118] In some embodiments, the heavy chain comprises a constant heavy chain sequence selected from the group consisting of the sequences set forth in SEQ ID NOs: 192-235 and 251-407.

[0119] In certain aspects, described herein is an isolated antibody that binds interleukin (IL)-4 receptor alpha (IL-4Rα), the antibody comprising: a) a variable heavy (VH) chain sequence having three heavy chain CDR sequences, CDR-H1, CDR-H2, and CDR-H3; and b) a variable light (VL) chain sequence having three light chain CDR sequences, CDR-L1, CDR-L2, and CDR-L3; wherein the antibody comprises: CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 66, or 187; CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 26 or 100; CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: NO:41 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:53; and wherein the heavy chain comprises a constant heavy chain sequence selected from the group consisting of SEQ ID NOs:192-235 and 251-407. In some embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO:113. In some embodiments, the antibody comprises the VL sequence set forth in SEQ ID NO:146.

[0120] In some embodiments, the light chain comprises a constant light chain sequence comprising the sequence set forth in SEQ ID NO:236.

[0121] In some embodiments, the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions increase one or more of antibody half-life, ADCC activity, ADCP activity, and / or CDC activity compared to an Fc region without the one or more substitutions.

[0122] In some embodiments, the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in reduced ADCC activity, ADCP activity, and / or CDC activity compared to an Fc region without the one or more substitutions.

[0123] In some embodiments, the one or more amino acid substitutions result in an increased half-life of the antibody compared to an antibody comprising a wild-type Fc region.

[0124] In some embodiments, the one or more amino acid substitutions are selected from the group consisting of S228P(SP), M252Y, S254T, T256E, T256D, T250Q, H285D, T307A, T307Q, T307R, T307W, L309D, Q411H, Q311V, A378V, E380A, M428L, N434A, N434S, N297A, D265A, L234A, L235A, and N434W. In some embodiments, the one or more amino acid substitutions are a combination of amino acid substitutions selected from the group consisting of: M428L / N434S (LS); M252Y / S254T / T256E (YTE); T250Q / M428L; T307A / E380A / N434A; T256D / T307Q (DQ); T256D / T307W (DW); M252Y / T256D (YD); T307Q / Q311V / A378V (QVV); T256D / H285D / T307R / Q311V / A378V(DDRVV); L309D / Q311H / N434S(DHS); S228P / L235E(SPLE); L234A / L235A, M428L / N434A, L234A / G237A, L234A / L235A / G237A, L234A / L235A / P329G, D265A / YTE, LALA / YTE, LAGA / YTE, LAL AGA / YTE, LALAPG / YTE, N297A / LS; D265A / LS; LALA / LS; LA LAGA / LS;LALAPG / LS;N297A / DHS;D265A / DHS;LALA / DHS;LAGA / DHS;LALAGA / DHS;LALAPG / DHS;SP / YTE;SPLE / YTE;SP / LS;S PLE / LS, SP / DHS; SPLE / DHS; N297A / LA; D265A / LA, LALA / LA, LAGA / LA, LALAGA / LA, LALAPG / LA, N297A / N434A; D265A / N434A;LALA / N434A, LAGA / N434A, LALAGA / N434A, LALAPG / N434A, N297A / N434W, D265A / N434W, LALA / N434W, LAGA / N434W, LALAGA / N434W, LALAPG / N434W, N297A / DQ, D265A / DQ, LALA / DQ, LAGA / DQ, LALAGA / DQ, LALAPG / DQ, N297A / DW, D265A / DW, LALA / DW, LAGA / DW, LALAGA / DW, LALAPG / DW N297A / YD, D265A / YD, LALA / YD, LAGA / YD, LAL AGA / YD, LALAPG / YD, T307Q / Q311V / A378V(QVV), N297A / QV V, D265A / QVV, LALA / QVV, LAGA / QVV, LALAGA / QVV, LAL APG / QVV, DDRVV, N297A / DDRVV, D265A / DDRVV, LALA / DD RVV, LAGA / DDRVV, LALAGA / DDRVV and LALAPG / DDRVV. ;

[0125] In some embodiments, the Fc region binds to the neonatal Fc receptor (FcRn).

[0126] In some embodiments, the Fc region binds FcRn with a higher affinity at pH 6.0 than an antibody comprising a wild-type Fc region.

[0127] In some embodiments, the Fc region is expressed at a pH of <1 x 10 -7 The KD of M binds to FcRn.

[0128] In some embodiments, the antibody is a monoclonal antibody.

[0129] In some embodiments, the antibody binds to the IL-4Rα sequence shown in SEQ ID NOs: 237-240.

[0130] In some embodiments, the antibody has a specific activity at less than or equal to about 1 x 10 -9 M, 2x 10 -9 M, 3x 10 -9 M, 4x 10 -9 M, 5x 10 -9 M, 6x 10 -9 M, 7x 10 -9 M, 8x 10 -9M, 9x 10 -9 The KD of M binds to the IL-4Rα sequence shown in SEQ ID NOs: 237-240.

[0131] In some embodiments, the antibody has a specific activity at less than or equal to about 1 x 10 -10 The KD of M binds to the IL-4Rα sequence shown in SEQ ID NOs: 237-240.

[0132] In some embodiments, the antibody has a specific activity at less than or equal to about 1 x 10 -9 The KD of M binds to human IL-4Rα.

[0133] In some embodiments, the antibody exhibits a melting temperature greater than 68°C as measured by differential scanning fluorimetry (DSF).

[0134] In some embodiments, the antibody exhibits a melting temperature greater than 75°C as measured by differential scanning fluorimetry (DSF).

[0135] In some embodiments, the antibody exhibits an aggregation temperature equal to or greater than 71.2°C as measured by differential scanning fluorimetry (DSF).

[0136] In some embodiments, the antibody has a retention time of 15.2 minutes or less as measured by hydrophobic interaction chromatography.

[0137] In some embodiments, the antibody does not have the heavy chain variable region sequence shown in SEQ ID NO.113.

[0138] In some embodiments, the antibody does not have the heavy chain sequence shown in SEQ ID NO.146.

[0139] In certain embodiments, the separated antibodies are used to treat inflammatory disorders or diseases. In certain embodiments, the separated antibodies are used to treat atopic dermatitis. In certain embodiments, the separated antibodies are used to treat idiopathic pulmonary fibrosis. In certain embodiments, the separated antibodies are used to treat alopecia areata. In certain embodiments, the separated antibodies are used to treat chronic sinusitis with nasal polyps. In certain embodiments, the separated antibodies are used to treat chronic sinusitis without nasal polyps (CRSsNP). In certain embodiments, the separated antibodies are used to treat eosinophilic esophagitis (EoE). In certain embodiments, the separated antibodies are used to treat eosinophilic gastrointestinal disorders or diseases (ENID) selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC) and eosinophilic gastroenteritis (EGE). In certain embodiments, the isolated antibodies are used to treat Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA). In certain embodiments, the isolated antibodies are used to treat prurigo nodularis (PN). In certain embodiments, the isolated antibodies are used to treat chronic spontaneous urticaria (CSU). In certain embodiments, the isolated antibodies are used to treat chronic pruritus of unknown cause (CPUO). In certain embodiments, the isolated antibodies are used to treat bullous pemphigoid (BP). In certain embodiments, the isolated antibodies are used to treat cold-induced urticaria (ColdU). In certain embodiments, the isolated antibodies are used to treat allergic fungal sinusitis (AFRS). In certain embodiments, the isolated antibodies are used to treat allergic bronchopulmonary aspergillosis (ABPA). In certain embodiments, the isolated antibodies are used to treat chronic obstructive pulmonary disease (COPD). In certain embodiments, the isolated antibodies are used to treat inflammatory bowel disease, such as Crohn's disease or ulcerative colitis. In certain embodiments, the isolated antibodies are used to treat lupus. In certain embodiments, the isolated antibodies are used to treat rheumatoid arthritis.

[0140] In certain aspects, disclosed herein is an isolated polynucleotide or group of polynucleotides encoding the antibody, VH, VL, light chain, heavy chain, or antigen-binding portion thereof of any one of the above claims, and optionally wherein the polynucleotide or group of polynucleotides comprises cDNA.

[0141] In certain aspects, disclosed herein is a vector or set of vectors comprising a polynucleotide or set of polynucleotides disclosed herein.

[0142] In certain aspects, disclosed herein is a host cell comprising a polynucleotide or set of polynucleotides or a vector or set of vectors disclosed herein.

[0143] In certain aspects, disclosed herein is a method of producing an antibody, the method comprising expressing the antibody using a host cell disclosed herein and isolating the expressed antibody.

[0144] In certain aspects, disclosed herein is a pharmaceutical composition comprising an antibody according to any one of the embodiments disclosed herein and a pharmaceutically acceptable excipient.

[0145] In certain aspects, disclosed herein is a kit comprising an antibody according to any one of the embodiments disclosed herein or a pharmaceutical composition disclosed herein and instructions for use.

[0146] In certain aspects, described herein is a method for treating an inflammatory condition or disease in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody described herein or a pharmaceutical composition described herein. In certain embodiments of the methods described herein, the inflammatory condition or disease is atopic dermatitis. In certain embodiments, the inflammatory condition or disease is asthma. In certain embodiments, the inflammatory condition or disease is idiopathic pulmonary fibrosis. In certain embodiments of the methods described herein, the inflammatory condition or disease is alopecia areata. In certain embodiments, the inflammatory condition or disease is chronic sinusitis with nasal polyps. In certain embodiments, the inflammatory condition or disease is chronic sinusitis without nasal polyps (CRSsNP). In certain embodiments, the inflammatory condition or disease is eosinophilic esophagitis (EoE). In certain embodiments, the inflammatory condition or disease is an eosinophilic gastrointestinal disorder or disease (ENID) selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC), and eosinophilic gastroenteritis (EGE). In certain embodiments, the inflammatory condition or disease is Chargar-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA). In certain embodiments, the inflammatory condition or disease is prurigo nodularis (PN). In certain embodiments, the inflammatory condition or disease is chronic spontaneous urticaria (CSU). In certain embodiments, the inflammatory condition or disease is chronic pruritus of unknown cause (CPUO). In certain embodiments, the inflammatory condition or disease is bullous pemphigoid (BP). In certain embodiments, the inflammatory condition or disease is cold-induced urticaria (ColdU). In certain embodiments, the inflammatory condition or disease is allergic fungal sinusitis (AFRS). In certain embodiments, the inflammatory condition or disease is allergic bronchopulmonary aspergillosis (ABPA). In certain embodiments, the inflammatory condition or disease is chronic obstructive pulmonary disease (COPD). In certain embodiments, the inflammatory condition or disease is inflammatory bowel disease, such as Crohn's disease or ulcerative colitis. In certain embodiments, the inflammatory condition or disease is lupus. In certain embodiments, the inflammatory condition or disease is rheumatoid arthritis.

[0147] In certain aspects, disclosed herein is a method for treating a pathology associated with elevated IL-4Rα levels in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody according to any one of the embodiments disclosed herein or a pharmaceutical composition disclosed herein.

[0148] In certain aspects, disclosed herein is a method of reducing the biological activity of IL-4Rα in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody of any one of the embodiments disclosed herein or a pharmaceutical composition disclosed herein.

[0149] In certain aspects, disclosed herein is a method of inhibiting a TH2-type allergic response in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody according to any one of the embodiments disclosed herein or a pharmaceutical composition disclosed herein.

[0150] In certain aspects, disclosed herein is a method of preventing an inflammatory disorder or disease in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody of any one of the embodiments disclosed herein or a pharmaceutical composition disclosed herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0151] Figure 1A and Figure 1B The ability of exemplary antibodies and dupilumab to inhibit IL-4 and IL-13 binding is shown.

[0152] Figure 2A and Figure 2B The ability of exemplary antibodies and dupilumab to inhibit IL-4-induced and IL-13-induced STAT6 phosphorylation (pSTAT6) is demonstrated.

[0153] Figure 3A and Figure 3B The ability of exemplary antibodies and dupilumab to inhibit IL-4 and / or IL-13-induced TARC secretion is demonstrated.

[0154] Figure 4A and Figure 4B The ability of exemplary antibodies and dupilumab to inhibit IL-4-induced and IL-13-induced proliferation of TF-1 cells is shown.

[0155] Figure 5A and Figure 5B demonstrated that a single intravenous dose of Figure 5A ) or subcutaneous ( Figure 5B ) Serum concentrations of exemplary antibodies (Construct 13 (mAb422), Construct 38 (mAb471), and dupilumab) over time following administration of 25 mg / kg. DETAILED DESCRIPTION

[0156] definition

[0157] Unless otherwise defined, all technical terms, symbols and other scientific terms used herein are intended to have the meaning commonly understood by those skilled in the art. In some cases, for clarity and / or for ease of reference, the term with commonly understood meaning is defined herein, and including such definition herein should not necessarily be interpreted as representing the difference commonly understood in the art. Those skilled in the art generally fully understand that the technology and procedures described herein or quoted and generally use conventional methods to adopt, such as Sambrook et al., Molecular Cl oning:A Laboratory Manual the 4th edition (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY described in the molecular cloning method of extensive utilization. When appropriate, unless otherwise noted, the program relating to the use of commercially available test kits and reagents is generally carried out according to the scheme and conditions defined by manufacturers.

[0158] As used herein, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated.

[0159] It is to be understood that aspects and embodiments of the present invention described herein include "comprising," "consisting of," and "consisting essentially of" aspects and embodiments.

[0160] For all compositions described herein and all methods of using the compositions described herein, the compositions may comprise the listed components or steps, or may "consist essentially of" the listed components or steps. When a composition is described as "consisting essentially of" the listed components, the composition contains the listed components and may contain other components that do not substantially affect the condition being treated, but does not contain any other components other than those specifically listed that substantially affect the condition being treated; or if a composition contains additional components other than those listed that substantially affect the condition being treated, the composition does not contain the additional components in sufficient concentration or amount to substantially affect the condition being treated. When a method is described as "consisting essentially of" the listed steps, the method contains the listed steps and may contain other steps that do not substantially affect the condition being treated, but the method does not contain any other steps other than those specifically listed that substantially affect the condition being treated. As a non-limiting specific example, when a composition is described as 'consisting essentially of' an ingredient, the composition may additionally contain any amount of pharmaceutically acceptable carriers, vehicles or diluents and other such ingredients that do not substantially affect the condition being treated.

[0161] As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which they have been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors."

[0162] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acids have been introduced and the progeny of such cells. Host cells include "transformants" (or "transformed cells") and "transfectants" (or "transfected cells"), each of which includes the primary transformed or transfected cell and the progeny derived therefrom. The nucleic acid content of such progeny may not be exactly the same as that of the parent cell and may contain mutations. "Recombinant host cell" or "host cell" refers to a cell that includes an exogenous polynucleotide, regardless of the method used for insertion (e.g., direct uptake, transduction, f-mating, or other methods known in the art to produce recombinant host cells).

[0163] As used herein, the term "eukaryote" refers to organisms belonging to the lineage domain Eukaryota, such as animals (including but not limited to mammals, insects, reptiles, birds, etc.), ciliates, plants (including but not limited to monocots, dicots, algae, etc.), fungi, yeasts, flagellates, microsporidia, protists, etc.

[0164] As used herein, the term "prokaryotic organism" refers to a prokaryotic organism. For example, the non-eukaryotic organism can belong to the Eubacteria (including but not limited to Escherichia coli, Thermus thermophilus, Bacillus stearothermophilus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, etc.) lineage domain, or the Archaea (including but not limited to Methanococcus jannaschii, Methanobacterium thermoautotrophicum, Halobacterium such as Haloferax volcanii and Halobacterium species NRC-1, Archaeoglobus fulgidus, Pyrococcus furi osus, Pyrococcus horikoshii, etc.). horikoshii), Aeuropyru m pernix, etc.) phylogenetic domain.

[0165] As used herein, an "effective amount" or "therapeutically effective amount" refers to an amount of a therapeutic compound (e.g., an anti-IL-4R antibody) administered to a subject as a single dose or as part of a series of doses (alone or in combination with another therapeutic modality) that is effective to produce or promote a desired therapeutic effect. Examples of desired therapeutic effects are reduction of an aberrant immune response, slowing or delaying disease progression, stabilization of the disease, and amelioration of one or more symptoms. An effective amount can be given in one or more doses.

[0166] The term "treating" (and variations such as "treat" or "treatment") refers to clinical intervention intended to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed during the course of clinical pathology. Desirable effects of treatment include preventing recurrence of the disease, alleviating symptoms, alleviating any direct or indirect pathological consequences of the disease, slowing the rate of disease progression, ameliorating or palliating the disease state, and alleviating or improving prognosis.

[0167] The term "sufficient amount" means an amount sufficient to produce a desired effect, such as an amount sufficient to modulate an immune response in a subject.

[0168] As used herein, the term "subject" or "individual" refers to a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cattle, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is human. In some embodiments, the subject suffers from a disease or condition treatable with an antibody provided herein. In some aspects, the disease or condition is cancer. In some aspects, the disease or condition is a viral infection.

[0169] The term "in vitro" refers to processes that occur in living cells grown separately from a living organism, for example, in tissue culture.

[0170] The term "in vivo" refers to processes that occur in a living organism.

[0171] The term "package insert" is used to refer to instructions customarily included in commercial packages of therapeutic or diagnostic products (e.g., kits), that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and / or warnings concerning the use of such therapeutic or diagnostic products.

[0172] The term "pharmaceutical composition" refers to a preparation that is in such form as to permit the biological activity of the active ingredient contained therein to be effective in treating a subject and that contains no additional components that are unacceptably toxic to the subject in the amounts provided in the pharmaceutical composition.

[0173] The terms "co-administration," "co-administer," and "combination" include the administration of two or more therapeutic agents simultaneously, concurrently, or sequentially within unspecified time limits. In one embodiment, the agents are present in a cell or subject simultaneously, or exert their biological or therapeutic effects simultaneously. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, the first agent may be administered prior to the administration of the second therapeutic agent.

[0174] The terms "modulate" and "modulation" refer to the decrease or inhibition, or the activation or increase of the recited variable.

[0175] The terms "increase" and "activate" refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold or more of the recited variable.

[0176] The terms "reduce" and "inhibit" refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold or more decrease in the recited variable.

[0177] The term "about" indicates and encompasses a specified value and a range above and below the value. In certain embodiments, the term "about" indicates a specified value ± 10%, ± 5%, or ± 1%. In certain embodiments, where applicable, the term "about" indicates a specified value ± one standard deviation of the value.

[0178] The term "agonism" refers to activation of receptor signaling to induce a biological response associated with activation of the receptor. An "agonist" is an entity that binds to and agonizes a receptor.

[0179] The term "antagonize" refers to the inhibition of receptor signaling to suppress the biological response associated with activation of the receptor. An "antagonist" is an entity that binds to and antagonizes a receptor.

[0180] For any of the structural and functional characteristics described herein, methods of determining these characteristics are known in the art.

[0181] The term "optionally" when used in sequence is meant to include one to all of the listed combinations and encompasses all sub-combinations.

[0182] The term "amino acid" refers to, for example, the twenty common naturally occurring amino acids. Naturally occurring amino acids include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine ​​(Cys; C), glutamic acid (Glu; E), glutamic acid (Gln; Q), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

[0183] The term "affinity" refers to the strength of the total non-covalent interaction between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or epitope). Unless otherwise indicated, as used herein, "affinity" refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen or epitope).

[0184] As used herein, the term "kd" (sec-1) refers to the dissociation rate constant for a specific antibody-antigen interaction. This value is also known as the koff value.

[0185] As used herein, the term "ka" (M-1 x sec-1) refers to the association rate constant for a specific antibody-antigen interaction. This value is also known as the kon value.

[0186] As used herein, the term "KD" (M) refers to the dissociation equilibrium constant for a particular antibody-antigen interaction. KD = kd / ka. In some embodiments, the affinity of an antibody is described in terms of the KD of the interaction between this antibody and its antigen. For clarity, as is known in the art, smaller KD values ​​indicate higher affinity interactions, while larger KD values ​​indicate lower affinity interactions.

[0187] As used herein, the term "KA" (M-1) refers to the association equilibrium constant for a specific antibody-antigen interaction. KA = ka / kd.

[0188] The term "antibody" is used herein in its broadest sense and includes certain types of immunoglobulin molecules that comprise one or more antigen binding domains that specifically bind to an antigen or epitope. Antibodies particularly include intact antibodies (e.g., intact immunoglobulins), antibody fragments, and multispecific antibodies.

[0189] An "anti-IL-4Rα antibody," "IL-4Rα antibody," or "IL-4Rα-specific antibody" is an antibody provided herein that specifically binds to the antigen IL-4Rα.

[0190] The term "epitope" means the portion of an antigen to which an antibody specifically binds.

[0191] As used herein, the term "hypervariable region" or "HVR" refers to the regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops ("hypervariable loops").

[0192] The term "antigen binding domain" means the portion of an antibody that is capable of specifically binding to an antigen or epitope.

[0193] The term "chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0194] The term "human antibody" or "fully human antibody" refers to an antibody having an amino acid sequence corresponding to an antibody produced by a human or human cell, or an amino acid sequence derived from a non-human source utilizing a human antibody repertoire or human antibody coding sequence (e.g., obtained from a human source or redesigned). Human antibodies particularly exclude humanized antibodies.

[0195] The term "humanized antibody" refers to a protein that has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions and / or additions, such that when administered to a human subject, the humanized antibody is less likely to induce an immune response and / or induces a less severe immune response than the non-human species antibody.

[0196] The term "multispecific antibody" refers to an antibody comprising two or more different antigen-binding domains that collectively specifically bind to two or more different epitopes.

[0197] A "monospecific antibody" is an antibody that contains one or more binding sites that specifically bind to a single epitope. An example of a monospecific antibody is a naturally occurring IgG molecule, which, although bivalent (i.e., having two antigen-binding domains), recognizes the same epitope at each of the two antigen-binding domains. Binding specificities can exist at any suitable valency.

[0198] The term "monoclonal antibody" refers to an antibody from a population of substantially homogeneous antibodies. The population of substantially homogeneous antibodies comprises antibodies that are substantially similar and that bind to the same epitope, except for variants that may typically occur during the production of monoclonal antibodies. Such variants are typically present only in small quantities. Monoclonal antibodies are typically obtained by methods including selecting a single antibody from a variety of antibodies. For example, a selection method can be used to select a unique clone from a variety of clones, such as a group of hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones. The selected antibody can be further modified to, for example, improve affinity for the target ("affinity maturation"), humanize the antibody, improve its yield in cell culture, and / or reduce its immunogenicity in a subject.

[0199] The term "single chain" refers to a molecule comprising amino acid monomers linearly connected via peptide bonds. In a specific embodiment of this invention, in a single-chain Fab molecule, the C-terminus of the Fab light chain is connected to the N-terminus of the Fab heavy chain. As described in more detail herein, scFv has a light chain (VL) variable domain connected to the N-terminus of the variable domain of the heavy chain (VH) from its C-terminus via a polypeptide chain. Alternatively, scFv comprises a polypeptide chain in which the C-terminus of VH is connected to the N-terminus of VL via a polypeptide chain.

[0200] A "Fab fragment" (also called fragment antigen binding) contains the constant domain (CL) of the light chain and the first constant domain (CH1) of the heavy chain, as well as the variable domains VL and VH on the light and heavy chains, respectively. The variable domains contain the complementarity determining loops (CDRs, also called hypervariable regions (HVRs)) involved in antigen binding. Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region.

[0201] "F(ab')2" fragments contain two Fab' fragments joined by a disulfide bond near the hinge region. F(ab')2 fragments can be produced, for example, by recombinant methods or by pepsin digestion of intact antibodies. F(ab') fragments can be dissociated, for example, by treatment with β-mercaptoethanol.

[0202] An "Fv" fragment comprises a non-covalently linked dimer of one heavy- and one light-chain variable domain.

[0203] "Single-chain Fv" or "sFv" or "scFv" includes the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. In one embodiment, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding. For a review of scFv, see Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, Rosenburg and Moore, eds., Springer-Verlag, New York, pp. 269-315 (1994). HER2 antibody scFv fragments are described in WO93 / 16185; U.S. Pat. No. 5,571,894; and U.S. Pat. No. 5,587,458.

[0204] "scFv-Fc" fragments comprise an scFv connected to an Fc domain. For example, the Fc domain can be connected to the C-terminus of the scFv. The Fc domain can be after the VH or VL, depending on the orientation of the variable domains in the scFv (i.e., VH-VL or VL-VH). Any suitable Fc domain known in the art or described herein can be used. In some cases, the Fc domain comprises an IgG4 Fc domain.

[0205] The term "single domain antibody" or "sdAb" refers to a molecule in which one variable domain of an antibody specifically binds to an antigen in the absence of other variable domains. Single domain antibodies and fragments thereof are described in Arabi Ghahroudi et al. (1998) FEBS Letters 414:521-526 and Muyldermans et al. (2001) Trends in Biochem. Sci. 26:230-245, each of which is incorporated by reference in its entirety. Single domain antibodies are also referred to as sdAbs or nanobodies. sdAbs are extremely stable and easily expressed with the Fc chain of an antibody as a fusion partner (Harmsen MM, De Haard HJ (2007) "Properties, production, and applications of camelid single-domain antibody fragments". Appl. Microbiol Biotechnol. 77 (1): 13-22).

[0206] The terms "full-length antibody," "intact antibody," and "complete antibody" are used interchangeably herein to refer to an antibody that has a structure substantially similar to a naturally occurring antibody structure and has heavy chains that include an Fc region. For example, when used to refer to an IgG molecule, a "full-length antibody" is an antibody that comprises two heavy chains and two light chains.

[0207] The term "antibody fragment" refers to an antibody comprising a portion of an intact antibody, such as the antigen-binding region or variable region of an intact antibody. Antibody fragments include, for example, Fv fragments, Fab fragments, F(ab')2 fragments, Fab' fragments, scFv (sFv) fragments, and scFv-Fc fragments.

[0208] The term "Fc domain" or "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions.

[0209] The term "substantially purified" refers to a construct described herein or variants thereof that are substantially or essentially free of components that normally accompany or interact with the protein as found in its naturally occurring environment (i.e., native cells; or, in the case of recombinantly produced antibodies, host cells, which, in certain embodiments, are essentially free of cellular material), and includes preparations of protein having less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% (by dry weight) of contaminating protein.

[0210] In the context of two or more nucleic acid or polypeptide sequences, the term "percent identity" refers to two or more sequences or subsequences that have a specified percentage of identical nucleotides or amino acid residues when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., using publicly available computer software such as BLAST, BLASTP, BLASTN, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software, or other algorithms available to those skilled in the art), or by visual inspection. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). One skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required to achieve maximum alignment over the full length of the sequences being compared. Depending on the application, the percentage "identity" can be present over a region of the sequences being compared, such as a functional domain, or over the full length of the two sequences being compared.

[0211] For sequence comparison, usually a sequence serves as a reference sequence compared to a test sequence. When using a sequence comparison algorithm, the test sequence and the reference sequence are input into a computer, subsequence coordinates are specified if necessary, and sequence algorithm program parameters are specified. The sequence comparison algorithm then calculates the sequence identity percentage of the test sequence relative to the reference sequence based on the specified program parameters.

[0212] Sequences for comparison can be optimally aligned, for example, by the local homology algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2:482; the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443; the search similarity method of Pearson and Lipman (1988) Proc. Nat'l. Acad. Sci. USA 85:2444; computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.); or by visual inspection (see generally, Ausubel et al., supra).

[0213] An example of an algorithm suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov / ).

[0214] 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50.

[0215] It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0216] Anti-IL-4Rα antibody

[0217] Antibody structure

[0218] The present application provides antibodies and compositions comprising antibodies that bind to IL-4 receptor alpha (IL-4Rα).

[0219] The immunoglobulin genes identified include kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes and a variety of immunoglobulin variable region genes. Light chains are classified as kappa or lambda. The "class" of an antibody or immunoglobulin refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG and IgM, and several of these antibodies can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are referred to as α, δ, epsilon, gamma and mu, respectively.

[0220] An exemplary immunoglobulin (antibody) structural unit is composed of two pairs of polypeptide chains, each pair having a "light" chain (about 25kD) and a "heavy" chain (about 50-70kD). The N-terminal domain of each chain defines a variable region of about 100 to 110 or more amino acids, which is primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chain domains, respectively. The IgG1 heavy chain comprises VH, CH1, CH2, and CH3 domains from N-terminus to C-terminus, respectively. The light chain comprises VL and CL domains from N-terminus to C-terminus. The IgG1 heavy chain comprises a hinge between the CH1 and CH2 domains. In certain embodiments, the immunoglobulin construct comprises at least one immunoglobulin domain from IgG, IgM, IgA, IgD, or IgE linked to a therapeutic polypeptide. In some embodiments, the immunoglobulin domains found in the antibodies provided herein are derived from or are derived from immunoglobulin-based constructs, such as bifunctional antibodies or nanobodies. In certain embodiments, the immunoglobulin constructs described herein comprise at least one immunoglobulin domain from a heavy chain antibody, such as a camelid antibody. In certain embodiments, the immunoglobulin constructs provided herein comprise at least one immunoglobulin domain from a mammalian antibody, such as a bovine antibody, a human antibody, a camel antibody, a mouse antibody, or any chimeric antibody.

[0221] In some embodiments, the antibodies provided herein comprise a heavy chain. In one embodiment, the heavy chain is IgA. In one embodiment, the heavy chain is IgD. In one embodiment, the heavy chain is IgE. In one embodiment, the heavy chain is IgG. In one embodiment, the heavy chain is IgM. In one embodiment, the heavy chain is IgG1. In one embodiment, the heavy chain is IgG2. In one embodiment, the heavy chain is IgG3. In one embodiment, the heavy chain is IgG4. In one embodiment, the heavy chain is IgA1. In one embodiment, the heavy chain is IgA2.

[0222] In some embodiments, the antibody is an IgG1 antibody. In some embodiments, the antibody is an IgG3 antibody. In some embodiments, the antibody is an IgG2 antibody. In some embodiments, the antibody is an IgG4 antibody.

[0223] Typically, a native four-chain antibody comprises six HVRs; three in VH (H1, H2, H3) and three in VL (L1, L2, L3). HVRs typically comprise amino acid residues from hypervariable loops and / or from complementary determining regions (CDRs), the latter having the highest sequence variability and / or involving antigen recognition. Except for CDR1 in VH, CDRs typically comprise amino acid residues forming hypervariable loops. Hypervariable regions (HVRs) are also referred to as "complementary determining regions" (CDRs), and these terms are used interchangeably herein with respect to the portion of the variable region that forms the antigen-binding region. This particular region has been described by Kabat et al., US Pat. of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and Chothia et al. (1987) J Mol Biol 196: 901-917, wherein each definition includes an overlap or subset of amino acid residues when compared to each other. However, the application of any definition of CDRs relating to antibodies or variants thereof is intended to be within the scope of the terms defined and used herein. The exact number of residues encompassing a particular CDR will vary depending on the sequence and size of the CDR. Given the variable region amino acid sequence of an antibody, one skilled in the art can routinely determine which residues comprise a particular CDR.

[0224] The amino acid sequence boundaries of a CDR can be determined by one skilled in the art using any of a variety of known numbering schemes, including those described by Kabat et al., supra ("Kabat" numbering scheme); Al-Lazikani et al. (1997) J. Mol. Biol. 273:927-948 ("Chothia" numbering scheme); MacCallum et al. (1996) J. Mol. Biol. 262:732-745 ("Contact" numbering scheme); Lefranc et al. (2003) Dev. Comp. Immunol. 27:55-77 ("IMGT" numbering scheme); and Honegge and Plückthun, (2001) J. Mol. Biol. 309:657-70 ("AHo" numbering scheme); each of which is incorporated herein by reference in its entirety.

[0225] Table 1 provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 identified by the Kabat and Chothia schemes. For CDR-H1, the residue numbering is provided using both the Kabat and Chothia numbering schemes.

[0226] CDRs can be assigned, for example, using antibody numbering software, such as Abnum available at www.bioinf.org.uk / abs / abnum / , and described in Abhinandan and Martin, (2008) Immunology 45:3832-3839, incorporated by reference in its entirety.

[0227] Table 1: Residues in CDRs according to Kabat and Chothia numbering schemes.

[0228]

[0229] *When numbering using the Kabat numbering convention, the C-terminus of CDR-H1 varies between H32 and H34, depending on the length of the CDR.

[0230] When referring to residues in the antibody heavy chain constant region, the "EU numbering scheme" is generally used (e.g., as reported in Kabat et al., supra). Unless otherwise stated, the EU numbering scheme is used to refer to residues in the antibody heavy chain constant region described herein.

[0231] One example of an antigen binding domain is an antigen binding domain formed by a VH-VL dimer of an antibody. Another example of an antigen binding domain is an antigen binding domain formed by diversification of certain loops from the tenth fibronectin type III domain of Adnectin. The antigen binding domain may include CDRs 1, 2, and 3 from the heavy chain, in the order described; and CDRs 1, 2, and 3 from the light chain, in the order described.

[0232] Epitopes often consist of surface-accessible amino acid residues and / or sugar side chains and may have specific three-dimensional structural features as well as charge-to-mass ratio characteristics. Conformational and non-conformational epitopes are distinguished in that binding to the former, but not the latter, may be lost in the presence of denaturing solvents. An epitope may contain amino acid residues that are directly involved in binding, and other amino acid residues that are not directly involved in binding. The epitope to which an antibody binds can be determined using known techniques for epitope determination, for example, by testing the binding of the antibody to IL-4Rɑ variants or chimeric IL-4Rɑ variants having different point mutations.

[0233] To screen for antibodies that bind to an epitope on the target antigen (e.g., IL-4Rα) to which the antibody of interest binds, a conventional cross-blocking assay, such as that described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, eds. Harlow and David Lane (1988), can be performed. Alternatively or additionally, epitope mapping can be performed by methods known in the art.

[0234] Chimeric antibodies are antibodies in which a portion of the heavy and / or light chain is derived from a particular source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0235] Human or fully human antibodies are antibodies having an amino acid sequence corresponding to an antibody produced by a human or human cell or an amino acid sequence derived from a non-human source utilizing a human antibody repertoire or human antibody coding sequence (e.g., obtained from a human source or redesigned). Human antibodies particularly exclude humanized antibodies.

[0236] Humanized antibodies have sequences that differ from those of antibodies derived from non-human species by one or more amino acid substitutions, deletions, and / or additions, such that when administered to human subjects, the humanized antibodies are less likely to induce an immune response and / or induce a less severe immune response than non-human species antibodies. In one embodiment, certain amino acids in the framework and constant domains of the heavy and / or light chains of non-human species antibodies are mutated to produce humanized antibodies. In another embodiment, the constant domains from human antibodies are fused to the variable domains of non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of non-human antibodies are altered to reduce the possible immunogenicity of the antibodies when the non-human antibodies are administered to human subjects, wherein the amino acid residues that are altered are not critical for the immunospecific binding of the antibody to its antigen, or the changes in the amino acid sequence performed are conservative changes such that the binding of the humanized antibody to the antigen is not significantly worse than that of the non-human antibody to the antigen. Examples of how to prepare humanized antibodies can be found in U.S. Patent Nos. 6,054,297; 5,886,152; and 5,877,293. For further details, see Jones et al. (1986) Nature 321:522-525; Riechmann et al. (1988) Nature 332:323-329; and Presta (1992) Curr. Op. Struct. Biol. 2:593-596, each of which is incorporated by reference in its entirety.

[0237] The two or more different epitopes can be on the same antigen (e.g., a single IL-4Rα) or on different antigens (e.g., different IL-4Rα molecules, or an IL-4Rα molecule and a non-IL-4Rα molecule). In some aspects, the multispecific antibody binds to two different epitopes (i.e., a "bispecific antibody"). In some aspects, the multispecific antibody binds to three different epitopes (i.e., a "trispecific antibody").

[0238] Anti-IL-4Rα antibodies may include antibodies described herein, such as clones shown in the figures and / or tables. In some embodiments, the antibodies comprise alternative scaffolds. In some embodiments, the antibodies consist of alternative scaffolds. In some embodiments, the antibodies consist essentially of alternative scaffolds. In some embodiments, the antibodies comprise antibody fragments. In some embodiments, the antibodies consist of antibody fragments. In some embodiments, the antibodies consist essentially of antibody fragments.

[0239] In some embodiments, the antibody is a monoclonal antibody.

[0240] In some embodiments, the antibody is a polyclonal antibody.

[0241] In some embodiments, the antibodies are produced by hybridomas. In other embodiments, the antibodies are produced by recombinant cells engineered to express the desired variable and constant domains.

[0242] In some embodiments, the antibody may be a single-chain antibody or other antibody derivative that retains antigen specificity and the lower hinge region or a variant thereof.

[0243] In some embodiments, the antibody may be a multifunctional antibody, a recombinant antibody, a fully human antibody, a humanized antibody, a fragment or a variant thereof. In a specific embodiment, the antibody fragment or derivative thereof is selected from the group consisting of a Fab fragment, a Fab'2 fragment, a CDR and a scFv.

[0244] Table 2. Sequences of IL-4Rα antibody constructs - vH, vL and associated CDRs

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254]

[0255]

[0256]

[0257]

[0258] V H domain

[0259] In some embodiments, the antibodies provided herein comprise a VH sequence selected from the group consisting of SEQ ID NOs: 113-145.

[0260] In some embodiments, the antibodies provided herein comprise a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the illustrative VH sequences provided in SEQ ID NOs: 113-145. H In some embodiments, the antibody of the present invention is a polypeptide or a sequence that has at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acid replacements. In some respects, the amino acid replacement is a conservative amino acid replacement. In some embodiments, the antibody described in this paragraph is referred to as " variant " in this article. In some embodiments, this type of variant is for example derived from the sequence provided herein by affinity maturation, site-directed mutagenesis, random mutagenesis or any other method known in the art or described herein. In some embodiments, this type of variant is not derived from the sequence provided herein, and can for example be separated again according to the method for obtaining antibody provided herein.

[0261] V L domain

[0262] In some embodiments, the antibodies provided herein comprise a VL sequence selected from the group consisting of SEQ ID NOs: 146-186.

[0263] In some embodiments, the antibodies provided herein comprise a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the illustrative VL sequences provided in SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the illustrative VL sequences provided in SEQ ID NOs: 146-186, having at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0264] VH-VL combination

[0265] In some embodiments, the antibodies provided herein comprise a VH sequence selected from the group consisting of SEQ ID NOs: 113-145; and a VL sequence selected from the group consisting of SEQ ID NOs: 146-186.

[0266] In certain aspects, any of SEQ ID NOs: 113-145 can be combined with any of SEQ ID NOs: 146-186.

[0267] In certain aspects, the antibodies provided herein comprise the VH and VL sequences of the constructs provided in Table 2.

[0268] In some embodiments, the antibodies provided herein comprise a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VH sequence provided in SEQ ID NOs: 113-145; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a VL sequence provided in SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise a VH sequence provided in SEQ ID NOs: 113-145 having at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid substitutions; and a VL sequence provided in SEQ ID NOs: 146-186 having at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0269] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 147. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 148. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 115 and the VL sequence set forth in SEQ ID NO: 149. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 150. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 116 and the VL sequence set forth in SEQ ID NO: 151. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 117 and the VL sequence set forth in SEQ ID NO: 152. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 118 and the VL sequence set forth in SEQ ID NO: 153. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 117 and the VL sequence set forth in SEQ ID NO: 154. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 119 and the VL sequence set forth in SEQ ID NO: 155. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 120 and the VL sequence set forth in SEQ ID NO: 156. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 120 and the VL sequence set forth in SEQ ID NO: 157. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 158. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 121 and the VL sequence set forth in SEQ ID NO: 158. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 122 and the VL sequence set forth in SEQ ID NO: 158. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 159. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 123 and the VL sequence set forth in SEQ ID NO: 159. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 124 and the VL sequence set forth in SEQ ID NO: 160.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 125 and the VL sequence set forth in SEQ ID NO: 161. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 126 and the VL sequence set forth in SEQ ID NO: 162. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 127 and the VL sequence set forth in SEQ ID NO: 163. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 128 and the VL sequence set forth in SEQ ID NO: 164. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 124 and the VL sequence set forth in SEQ ID NO: 158. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 129 and the VL sequence set forth in SEQ ID NO: 165. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 130 and the VL sequence set forth in SEQ ID NO: 166. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 131 and the VL sequence set forth in SEQ ID NO: 167. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 132 and the VL sequence set forth in SEQ ID NO: 159. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 133 and the VL sequence set forth in SEQ ID NO: 159. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 121 and the VL sequence set forth in SEQ ID NO: 168. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 134 and the VL sequence set forth in SEQ ID NO: 169. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 113 and the VL sequence set forth in SEQ ID NO: 170. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 134 and the VL sequence set forth in SEQ ID NO: 171. In certain embodiments, the antibody comprises a VH sequence as set forth in SEQ ID NO: 135 and a VL sequence as set forth in SEQ ID NO: 172. In certain embodiments, the antibody comprises a VH sequence as set forth in SEQ ID NO: 136 and a VL sequence as set forth in SEQ ID NO: 173. In certain embodiments, the antibody comprises a VH sequence as set forth in SEQ ID NO: 131 and a VL sequence as set forth in SEQ ID NO: 174.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 137 and the VL sequence set forth in SEQ ID NO: 175. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 176. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 138 and the VL sequence set forth in SEQ ID NO: 177. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 134 and the VL sequence set forth in SEQ ID NO: 178. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 139 and the VL sequence set forth in SEQ ID NO: 179. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 140 and the VL sequence set forth in SEQ ID NO: 180. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 141 and the VL sequence set forth in SEQ ID NO: 181. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 142 and the VL sequence set forth in SEQ ID NO: 182. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 143 and the VL sequence set forth in SEQ ID NO: 183. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 114 and the VL sequence set forth in SEQ ID NO: 184. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 144 and the VL sequence set forth in SEQ ID NO: 185. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 145 and the VL sequence set forth in SEQ ID NO: 186.

[0270] In certain embodiments of any of the antibodies described above, the antibody further comprises a heavy chain comprising a human IgG sequence selected from the sequences shown in SEQ ID NOs: 192-235 and 251-407.

[0271] In certain embodiments, the antibody comprises a VH sequence as set forth in SEQ ID NO: 113; a VL sequence as set forth in SEQ ID NO: 146; and the antibody further comprises a heavy chain comprising a human IgG sequence selected from the group consisting of SEQ ID NOs: 192-235 and 251-407. In certain embodiments of any of the antibodies described above, the antibody further comprises a constant light chain sequence comprising the sequence set forth in SEQ ID NO: 236.

[0272] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 134, the VL sequence set forth in SEQ ID NO: 178, a heavy chain constant region comprising a LALA / YTE substitution, and a human kappa light chain constant region. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 134, the VL sequence set forth in SEQ ID NO: 178, a heavy chain constant region comprising the sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321, and a light chain constant region comprising the sequence set forth in SEQ ID NO: 236. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 134; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 178; a heavy chain constant region comprising a LALA / YTE substitution; and a human kappa light chain constant region. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 134; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 178; a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 205 or SEQ ID NO: 321; and a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 236 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: NO:236 has a light chain constant region with a sequence at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical.

[0273] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 130, the VL sequence set forth in SEQ ID NO: 166, a heavy chain constant region comprising a LALA / YTE substitution, and a human kappa light chain constant region. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 130, the VL sequence set forth in SEQ ID NO: 166, a heavy chain constant region comprising the sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321, and a light chain constant region comprising the sequence set forth in SEQ ID NO: 236. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 130; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 166; a heavy chain constant region comprising a LALA / YTE substitution; and a human kappa light chain constant region. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 130; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 166; a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 205 or SEQ ID NO: 321; and a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 236 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: NO:236 has a light chain constant region with a sequence at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical.

[0274] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 121, the VL sequence set forth in SEQ ID NO: 158, a heavy chain constant region comprising a LALA / YTE substitution, and a human kappa light chain constant region. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 121, the VL sequence set forth in SEQ ID NO: 158, a heavy chain constant region comprising the sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321, and a light chain constant region comprising the sequence set forth in SEQ ID NO: 236. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 121; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 158; a heavy chain constant region comprising a LALA / YTE substitution; and a human kappa light chain constant region. In certain embodiments, the antibody comprises: a VH sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VH sequence provided in SEQ ID NO: 121; and a VL sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the VL sequence provided in SEQ ID NO: 158; a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 205 or SEQ ID NO: 321 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 205 or SEQ ID NO: 321; and a heavy chain constant region comprising a sequence set forth in SEQ ID NO: 236 or a sequence that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: NO:236 has a light chain constant region with a sequence at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical.

[0275] CDR

[0276] In some embodiments, disclosed herein is an antibody comprising 1, 2, 3, 4, 5, or 6 of the CDRs in Table 2. In some embodiments, disclosed herein is an antibody comprising 6 Kabat CDRs in Table 2, 6 Chothia CDRs in Table 2, or 6 IMGT CDRs in Table 2.

[0277] In some embodiments, the antibodies provided herein comprise one to three CDRs of a VH domain selected from SEQ ID NOs: 113-145. In some embodiments, the antibodies provided herein comprise two to three CDRs of a VH domain selected from SEQ ID NOs: 113-145. H In some embodiments, the antibodies provided herein comprise three CDRs of the VH domain selected from SEQ ID NOs: 113-145. In some aspects, the CDRs are exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are IMGT CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs.

[0278] In some embodiments, the CDR is a CDR that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to a CDR-H1, CDR-H2 or CDR-H3 of SEQ ID NOs: 1-112 and 187-191. In some embodiments, the CDR-H1 is a CDR-H1 of a VH domain selected from SEQ ID NOs: 113-145 having at most 1, 2, 3, 4 or 5 amino acid substitutions. In some embodiments, the CDR-H2 is a CDR-H2 of a VH domain of SEQ ID NOs: 113-145 having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some embodiments, the CDR-H3 is a CDR-H3 selected from the VH domains of SEQ ID NO: 113-145, having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants". In some embodiments, such variants are derived from sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis or any other method known in the art or described herein. In some embodiments, such variants are not derived from sequences provided herein, and can be, for example, re-isolated according to the method for obtaining antibodies provided herein.

[0279] In some embodiments, the antibodies provided herein comprise one to three CDRs of the VL domains of SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise two to three CDRs of the VL domains of SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise three CDRs of the VL domains of SEQ ID NOs: 146-186. In some aspects, the CDRs are exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are IMGT CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs.

[0280] In some embodiments, the CDR is a CDR having at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the CDR-L1, CDR-L2 or CDR-L3 of SEQ ID NOs: 26-65 and 100-112. In some embodiments, the CDR-L1 is the CDR-L1 of the VL domain of SEQ ID NOs: 146-186, which has at most 1, 2, 3, 4 or 5 amino acid substitutions. In some embodiments, the CDR-L2 is the CDR-L2 of the VL domain of SEQ ID NOs: 41-52 and 108-112, which has at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some embodiments, the CDR-L3 is a CDR-L3 in the VL domain of SEQ ID NO:146-186, which has at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibody described in this paragraph is referred to as "variant" in this article. In some embodiments, such variants are derived from sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis or any other method known in the art or described herein. In some embodiments, such variants are not derived from sequences provided herein, and can be, for example, isolated again according to the method for obtaining antibodies provided herein.

[0281] In some embodiments, the antibodies provided herein comprise one to three CDRs of the VH domain selected from SEQ ID NOs: 113-145 and one to three CDRs of the VL domain selected from SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise two to three CDRs of the VH domain selected from SEQ ID NOs: 113-145 and two to three CDRs of the VL domain selected from SEQ ID NOs: 146-186. In some embodiments, the antibodies provided herein comprise three CDRs of the VH domain selected from SEQ ID NOs: 113-145 and three CDRs of the VL domain selected from SEQ ID NOs: 146-186. In some aspects, the CDRs are exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are IMGT CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs.

[0282] In some embodiments, the antibodies provided herein comprise a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99. In some aspects, the CDR-H3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99. In some embodiments, the CDR-H3 is a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99 having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0283] In some embodiments, the antibodies provided herein comprise a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70, and 187-191. In some aspects, the CDR-H1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70, and 187-191. In some embodiments, the CDR-H1 is a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70, and 187-191 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from sequences provided herein, and can be isolated again, for example, according to the methods for obtaining antibodies provided herein.

[0284] In some embodiments, the antibodies provided herein comprise a CDR-H2 selected from SEQ ID NOs: 5-16 and 71-90. In some aspects, the CDR-H2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-H2 selected from SEQ ID NOs: 5-16 and 71-90. In some embodiments, the CDR-H2 is a CDR-H2 selected from SEQ ID NOs: 5-16 and 71-90 having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0285] In some embodiments, the antibodies provided herein comprise a CDR-L3 selected from SEQ ID NOs: 53-65. In some aspects, the CDR-L3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the CDR-L3 of SEQ ID NOs: 53-65. In some embodiments, the CDR-L3 is a CDR-L3 selected from SEQ ID NOs: 53-65 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated again, for example, according to the methods provided herein for obtaining antibodies.

[0286] In some embodiments, the antibodies provided herein comprise a CDR-L2 selected from SEQ ID NOs: 41-52 and 108-112. In some aspects, the CDR-L2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-L2 selected from SEQ ID NOs: 41-52 and 108-112. In some embodiments, the CDR-L2 is a CDR-L2 selected from SEQ ID NOs: 41-52 and 108-112 having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0287] In some embodiments, the antibodies provided herein comprise a CDR-L1 selected from SEQ ID NOs: 26-40 and 100-107. In some aspects, the CDR-L1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-L1 selected from SEQ ID NOs: 26-40 and 100-107. In some embodiments, the CDR-L1 is a CDR-L1 selected from SEQ ID NOs: 26-40 and 100-107 having at most 1, 2, 3, 4, 5, 6, 7 or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as "variants." In some embodiments, such variants are derived from the sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from the sequences provided herein and can be isolated de novo, for example, according to the methods provided herein for obtaining antibodies.

[0288] In some embodiments, the antibodies provided herein comprise: a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99; a CDR-H2 selected from SEQ ID NOs: 5-16 and 71-90; a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70 and 187-191; a CDR-L3 selected from SEQ ID NOs: 53-65; a CDR-L2 selected from SEQ ID NOs: 41-52 and 108-112; and a CDR-L1 selected from SEQ ID NOs: 26-40 and 100-107. In some embodiments, the CDR-H3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99, the CDR-H2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H2 of SEQ ID NOs: 5-16 and 71-90, the CDR-H1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70, and 187-191, wherein said CDR-L3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-L3 selected from the group consisting of SEQ ID NOs: 53-65, said CDR-L2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-L2 selected from the group consisting of SEQ ID NOs: 41-52 and 108-112, and said CDR-L1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to a CDR-L1 selected from the group consisting of SEQ ID NOs: 26-40 and 100-107.In some embodiments, the CDR-H3 is a CDR-H3 selected from SEQ ID NOs: 17-25 and 92-99 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NOs: 5-16 and 71-90 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 selected from SEQ ID NOs: 1-4, 66-70, and 187-191 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 selected from SEQ ID NOs: 53-65 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 selected from SEQ ID NOs: NO: 41-52 and 108-112 CDR-L2, which has up to 1, 2, 3 or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 selected from SEQ ID NO: 26-40 and 100-107, which has up to 1, 2, 3, 4, 5 or 6 amino acid substitutions.

[0289] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 67, or 187; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 26 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 41 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 53. In some embodiments, the CDR-H3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the CDR-H3 of SEQ ID NO: 17 or 91, the CDR-H2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the CDR-H2 of SEQ ID NO: 5, 71, or 79, the CDR-H1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the CDR-H1 of SEQ ID NO: 1, 67, or 187, and the CDR-L3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the CDR-L3 of SEQ ID NO: NO:53 has a CDR-L3 that is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical, the CDR-L2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDR-L2 of SEQ ID NO:41 or 108, and the CDR-L1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDR-L1 of SEQ ID NO:26 or 100.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 17 or 91 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 5, 71, or 79 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 1, 67, or 187 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 53 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 41 or 108 having at most 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is the CDR-L1 of SEQ ID NO: The CDR-L1 of NO: 26 or 100 having at most 1, 2, 3, 4, 5 or 6 amino acid substitutions.

[0290] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 54.

[0291] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 62, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0292] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0293] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0294] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 56.

[0295] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0296] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 46 or 109; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 58.

[0297] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 59.

[0298] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 68, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0299] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0300] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0301] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0302] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0303] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0304] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 21 or 96; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0305] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 62.

[0306] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0307] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 23, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0308] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 56.

[0309] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 23 or 97; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0310] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20, 18, or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0311] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0312] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 36 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0313] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 37 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0314] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 12, 74, or 86; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0315] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 13, 71, or 87; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0316] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0317] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0318] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 66, or 187; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 38 or 105; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 48 or 111; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 63.

[0319] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 41 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 64.

[0320] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 49 or 112; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 63.

[0321] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0322] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 58.

[0323] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 50 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0324] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0325] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 67, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 67, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 40 or 107; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 52 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0326] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0327] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22, 18, or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 60.

[0328] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 65.

[0329] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 62.

[0330] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 55.

[0331] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0332] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 61.

[0333] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 57.

[0334] In some embodiments, the antibodies provided herein comprise: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 25 or 99; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 53.

[0335] In some embodiments, the CDR-H3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H3 of Table 2; the CDR-H2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H2 of Table 2, and the CDR-H1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a CDR-H1 of Table 2. wherein the CDR-L3 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDR-L3 of Table 2, the CDR-L2 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDR-L2 of Table 2, and the CDR-L1 is at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the CDR-L1 of Table 2. In some embodiments, the CDR-H3 is a CDR-H3 of Table 2 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of Table 2 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of Table 2 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of Table 2 having at most 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of Table 2 having at most 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of Table 2 having at most 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0336] In certain embodiments of any of the antibodies described above, the antibody further comprises a heavy chain comprising a human IgG sequence selected from the group consisting of SEQ ID NOs: 192-235 and 251-407. In certain embodiments of any of the antibodies described above, the antibody further comprises a constant light chain sequence comprising the sequence set forth in SEQ ID NO: 236. In certain embodiments of any of the antibodies described above, the antibody further comprises a heavy chain comprising a human IgG sequence selected from the group consisting of SEQ ID NO: 205 and SEQ ID NO: 321 and a constant light chain sequence comprising the sequence set forth in SEQ ID NO: 236.

[0337] In some embodiments, the antibody comprises: a CDR-H1 comprising a sequence as set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising a sequence as set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising a sequence as set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising a sequence as set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising a sequence as set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising a sequence as set forth in SEQ ID NO: 61. In certain embodiments, the antibody further comprises a heavy chain constant region (e.g., an IgG1 constant region) comprising a LALA / YTE substitution. In certain embodiments, the antibody comprises a heavy chain comprising a heavy chain constant region selected from the group consisting of SEQ ID NOs: 205 and 321. In certain embodiments, the antibody comprises a human kappa light chain constant region. In some embodiments, the antibody comprises a constant light chain sequence comprising the sequence shown in SEQ ID NO:236.

[0338] In some embodiments, the antibody comprises: a CDR-H1 comprising a sequence as set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising a sequence as set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising a sequence as set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising a sequence as set forth in any one of SEQ ID NOs: 36 or 102; a CDR-L2 comprising a sequence as set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising a sequence as set forth in SEQ ID NO: 61. In certain embodiments, the antibody further comprises a heavy chain constant region (e.g., an IgG1 constant region) comprising a LALA / YTE substitution. In certain embodiments, the antibody comprises a heavy chain comprising a heavy chain constant region selected from the group consisting of SEQ ID NOs: 205 and 321. In certain embodiments, the antibody comprises a human kappa light chain constant region. In some embodiments, the antibody comprises a constant light chain sequence comprising the sequence shown in SEQ ID NO:236.

[0339] In some embodiments, the antibody comprises: a CDR-H1 comprising a sequence as set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising a sequence as set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising a sequence as set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising a sequence as set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising a sequence as set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising a sequence as set forth in SEQ ID NO: 61. In certain embodiments, the antibody further comprises a heavy chain constant region (e.g., an IgG1 constant region) comprising a LALA / YTE substitution. In certain embodiments, the antibody comprises a heavy chain comprising a heavy chain constant region selected from the group consisting of SEQ ID NOs: 205 and 321. In certain embodiments, the antibody comprises a human kappa light chain constant region. In some embodiments, the antibody comprises a constant light chain sequence comprising the sequence shown in SEQ ID NO:236.

[0340] In some aspects, the amino acid substitution is a conservative amino acid substitution. In some embodiments, the antibodies described in the present disclosure are referred to herein as "variants" or "clones". In some embodiments, such variants or clones are derived from sequences provided herein, for example, by affinity maturation, site-directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants or clones are not derived from sequences provided herein, and can be isolated again, for example, according to the method for obtaining antibodies provided herein.

[0341] Fc region

[0342] The structures of the Fc regions of different immunoglobulins and the glycosylation sites contained therein are known in the art. See Schroeder and Cavacini, J. (2010) Allergy Clin. Immunol. 125: S41-52, which is incorporated by reference in its entirety. The Fc region may be a naturally occurring Fc region or a modified Fc region described in the art or elsewhere in this disclosure.

[0343] Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991. As used herein, an "Fc polypeptide" of a dimeric Fc refers to one of the two polypeptides that form the dimeric Fc domain, i.e., a polypeptide comprising the C-terminal constant regions of immunoglobulin heavy chains that are capable of stable self-association. For example, the Fc polypeptide of a dimeric IgG Fc comprises IgG CH2 and IgG CH3 constant domain sequences. Fc can have classes IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0344] The terms "Fc receptor" and "FcR" are used to describe receptors that bind to the Fc region of an antibody. For example, an FcR may be a native sequence human FcR. Typically, FcRs are those that bind IgG antibodies (gamma receptors) and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and alternative splicing forms of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibiting receptor"), both of which have similar amino acid sequences that differ primarily in their cytoplasmic domains. Other isotypes of immunoglobulins may also be bound by certain FcRs (see, for example, Janeway et al., ImmunoBiology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999)). The activating receptor FcγRIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor FcγRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (reviewed in (1997) Annu. Rev. Immunol. 15: 203-234). FcRs are reviewed in Ravetch and Kinet, (1991) Annu. Rev. Immunol 9: 457-92; Capel et al. (1994) Immunomethods 4: 25-34; and de Haas et al. (1995) J. Lab. Clin. Med. 126: 330-41. Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein. The term also includes the neonatal receptor FcRn, which is responsible for the transport of maternal IgG to the fetus (Guyer et al. (1976) J. Immunol. 117: 587; and Kim et al. (1994) J. Immunol. 24: 249).

[0345] Modifications in the CH2 domain may affect the binding of FcRs to Fc. Various amino acid modifications in the Fc region are known in the art to selectively alter the affinity of Fc for different Fcγ receptors. In some aspects, the Fc comprises one or more modifications that promote selective binding of Fc-γ receptors.

[0346] Exemplary mutations that alter FcR binding to Fc are listed below:

[0347] ●S298A / E333A / K334A, S298A / E333A / K334A / K326A (Lu et al. (2011) J ImmunolMethods 365(1-2):132-41);

[0348] F243L / R292P / Y300L / V305I / P396L, F243L / R292P / Y300L / L235V / P396L (Stavenhagen et al. (2007) Cancer Res. 67(18):8882-90; Nordstrom et al. (2011) Breast Cancer Res. 13(6):R123);

[0349] F243L (Stewart et al. (2011) Protein Eng Des Sel. 24(9):671-8.), S298A / E333A / K334A (Shields et al. (2001) J Biol Chem. 276(9):6591-604);

[0350] ●S239D / I332E / A330L, S239D / I332E (Lazar et al. (2006) Proc Natl Acad Sci U SA. 103(11):4005-10);

[0351] ●S239D / S267E, S267E / L328F (Chu et al. (2008) Mol Immunol. 45(15):3926-33);

[0352] ●S239D / D265S / S298A / I332E, S239E / S298A / K326A / A327H, G237F / S298A / A330L / I332E, S239D / I332E / S298A, S239D / K326E / A330L / I332E / S298A, G236A / S239D / D270L / I332E, S239E / S267E / H268D, L234F / S267E / N325L, G237F / V266L / S267D and other mutations listed in WO2011 / 120134 and WO2011 / 120135, which are incorporated herein by reference. Therapeutic Antibody Engineering (William R. Strohl and Lila M. Strohl, Woodhead Publishing series in Biomedicine No. 11, ISBN 1 907568 37 9, October 2012) lists mutations on page 283.

[0353] In some embodiments, the antibodies described herein include modifications to improve their ability to mediate effector functions. Such modifications are known in the art and include afucosylation or Fc engineering of affinity for activating receptors, primarily FCGR3a for ADCC and C1q for CDC. Table 3 below summarizes various designs reported in the literature for effector function engineering.

[0354] Methods for generating antibodies with little or no fucose at Fc glycosylation sites (Asn 297 EU numbering) without altering the amino acid sequence are well known in the art. The technology (ProBioGen AG) is based on introducing genes for enzymes in a cellular pathway that is biased towards fucose biosynthesis into cells used for antibody production. This prevents antibody-producing cells from adding the sugar "fucose" to N-linked antibody carbohydrate moieties. (von Horsten et al. (2010) Glycobiology. 2010 Dec;20(12):1607-18. Another method for obtaining antibodies with reduced fucosylation levels can be found in U.S. Patent No. 8,409,572, which teaches selecting a cell line capable of producing lower levels of fucosylation on antibodies for use in antibody production. The antibody may be completely afucosylated (meaning it contains no detectable fucose) or it may be partially afucosylated, meaning the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15%, or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.

[0355] Thus, in one embodiment, the antibodies described herein may comprise a dimeric Fc comprising one or more amino acid modifications conferring improved effector function as indicated in Table 3. In another embodiment, the antibodies may be afucosylated to improve effector function.

[0356] Table 3: CH2 domain and effector function engineering

[0357]

[0358] Fc modifications that reduce FcgR and / or complement binding and / or effector function are known in the art. Recent publications describe strategies for engineering antibodies with reduced or silent effector activity (see Strohl, WR (2009), Curr Opin Biotech 20: 685-691, and Strohl, WR and Strohl LM, "Antibody Fc engineering for optimal antibody performance", Therapeutic Antibody Engineering, Cambridge: Woodhead Publishing (2012), p. 225 to p. 249). These strategies include using IgG2 / IgG4 scaffolds or introducing mutations in the hinge or CH2 region of Fc to reduce effector function via glycosylation modification. For example, U.S. Patent Publication No. 2011 / 0212087 (Strohl), International Patent Publication No. WO 2006 / 105338 (Xencor), U.S. Patent Publication No. 2012 / 0225058 (Xencor), U.S. Patent Publication No. 2012 / 0251531 (Genentech), and Strop et al. ((2012) J. Mol. Biol. 420:204-219) describe specific modifications for reducing binding of FcgR or complement to Fc.

[0359] Specific non-limiting examples of amino acid modifications known to reduce FcgR or complement binding to Fc include those identified below in Table 4:

[0360] Table 4: Modifications used to reduce FcgR or complement binding to Fc

[0361]

[0362]

[0363] Methods for generating antibodies with little or no fucose at Fc glycosylation sites (Asn 297 EU numbering) without altering the amino acid sequence are well known in the art. The technology (ProBioGen AG) is based on the introduction of genes for enzymes of the cellular pathways that are biased towards fucose biosynthesis into cells for antibody production. This prevents antibody-producing cells from adding the sugar "fucose" to the N-linked antibody carbohydrate moiety. (von Horsten et al. (2010) Glycobiology 20 (12): 1607-18). Examples of cell lines capable of producing defucosylated antibodies include CHO-DG44 (see von Horsten et al. (2010), supra) or Lec13 CHO cells with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxose-4-hexose reductase (RMD), which lack protein fucosylation (see Ripka et al. (1986) Arch.Biochem.Biophys.249: 533-545; U.S. Patent Publication No. 2003 / 0157108; WO 2003 / 0157108). 2004 / 056312; each of which is incorporated by reference in its entirety) and gene knockout cell lines, such as α-1,6-fucosyltransferase gene or FUT8 gene knockout CHO cells (see Yamane-Ohnuki et al. (2004) Biotech. Bioeng. 87:614-622; Kanda et al. (2006) Biotechnol. Bioeng. 94:680-688; and WO 2003 / 085107; each of which is incorporated by reference in its entirety). Another approach to obtaining antibodies with reduced fucosylation levels can be found in U.S. Patent No. 8,409,572, which teaches the selection of cell lines capable of producing lower fucosylation levels on antibodies for antibody production.

[0364] Examples of cell lines capable of producing defucosylated antibodies include CHO-DG44 (see von Horsten et al. (2010), supra) or Lec13 CHO cells with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxose-4-hexose reductase (RMD), which lack protein fucosylation (see Ripka et al. (1986), supra; U.S. Patent Publication No. 2003 / 0157108; WO 2004 / 056312; each of which is incorporated by reference in its entirety) and gene knockout cell lines, such as α-1,6-fucosyltransferase gene or FUT8 gene knockout CHO cells (see Yamane-Ohnuki et al. (2004), supra; Kanda et al. (2006), supra; and WO 2003 / 085107; each of which is incorporated by reference in its entirety).

[0365] The antibody may be completely afucosylated, meaning that it contains no detectable fucose, or it may be partially afucosylated, meaning that the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15%, or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.

[0366] In some aspects, the antibodies provided herein comprise an IgG1 domain having reduced fucose content at position Asn 297 compared to naturally occurring IgG1 domains. Such Fc domains are known to have improved ADCC. See Shields et al. (2002) J. Biol. Chem. 277: 26733-26740, which is incorporated by reference in its entirety. In some aspects, such antibodies do not comprise any fucose at position Asn 297. The amount of fucose can be determined using any suitable method, such as that described in WO 2008 / 077546, which is incorporated by reference in its entirety.

[0367] In certain embodiments, the antibodies provided herein comprise an Fc region having one or more amino acid substitutions that improve ADCC, e.g., substitutions at one or more of positions 298, 333, and 334 of the Fc region. In some embodiments, the antibodies provided herein comprise an Fc region having one or more amino acid substitutions at positions 239, 332, and 330, as described in Lazar et al. (2006), supra, which is incorporated by reference in its entirety.

[0368] Other illustrative glycosylation variants that can be incorporated into the antibodies provided herein are described, for example, in U.S. Patent Publication Nos. 2003 / 0157108, 2004 / 0093621, 2003 / 0157108, 2003 / 0115614, 2002 / 0164328, 2004 / 0093621, 2004 / 0132140, 2004 / 0110704, 2004 / 0110282, 2004 / 0109865; International Patent Publication No. 200 No. 0 / 61739, No. 2001 / 29246, No. 2003 / 085119, No. 2003 / 084570, No. 2005 / 035586, No. 2005 / 035778, No. 2005 / 053742, No. 2002 / 031140; Okazaki et al. (2004) J. Mol. Biol., 2004, 336:1239-1249; and Yamane-Ohnuki et al. (2004), supra; each of which is incorporated by reference in its entirety.

[0369] In some embodiments, the antibodies provided herein comprise at least one galactose residue in the oligosaccharide attached to the Fc region of the Fc region. Such antibody variants may have improved CDC function. Examples of such antibody variants are described in, for example, WO 1997 / 30087, WO 1998 / 58964, and WO 1999 / 22764; each of which is incorporated by reference in its entirety.

[0370] In some embodiments, the antibodies provided herein comprise one or more alterations that improve or reduce CIq binding and / or CDC. See U.S. Patent No. 6,194,551; WO 99 / 51642; and Idusogie et al. (2000) J. Immunol., 164:4178-4184; each of which is incorporated by reference in its entirety.

[0371] In certain embodiments, the heavy chain comprises a constant heavy chain sequence selected from the sequence shown in SEQ ID NO: 192-235 and 251-407. In certain embodiments, the constant heavy chain sequence (e.g., a constant heavy chain sequence selected from SEQ ID NO: 192-235 and 251-306) further comprises a C-terminal lysine (SEQ ID NO: 307-407). Although the C-terminal lysine may be present in the corresponding coding sequence of the constant heavy chain region, it tends to be cleaved during manufacture or after administration. Therefore, provided herein are heavy chain constant region sequences with and without a C-terminal lysine.

[0372] In certain embodiments, the antibodies provided herein comprise the VH sequences and VL sequences provided in Table 2, and a heavy chain constant region selected from the group consisting of sequences shown in SEQ ID NOs: 192-235 and 251-407. H Sequence and V L The sequence is shown in SEQ ID NO: 146, and the heavy chain constant region and light chain constant region are shown in SEQ ID NO: 147.

[0373] In certain embodiments, the antibody comprises a V sequence as shown in SEQ ID NO: 113. H Sequence and V shown in SEQ ID NO: 146 L sequence; and wherein the constant heavy chain comprises a human IgG sequence selected from the group consisting of sequences shown in SEQ ID NOs: 192-235 and 251-407.

[0374] In certain embodiments, the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions increase one or more of antibody half-life, ADCC activity, ADCP activity, or CDC activity compared to an Fc region without the one or more substitutions. In certain embodiments, the one or more amino acid substitutions increase antibody half-life at pH 6.0 compared to an antibody comprising a wild-type Fc region. In certain embodiments, the increased half-life of the antibody is about 10,000-fold, 1,000-fold, 500-fold, 100-fold, 50-fold, 20-fold, 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4.5-fold, 4-fold, 3.5-fold, 3-fold, 2.5-fold, 2-fold, 1.95-fold, 1.9-fold, 1.85-fold, 1.8-fold, 1.75-fold, 1.7-fold, 1.65-fold, 1.6-fold, 1.55-fold, 1.50-fold, 1.45-fold, 1.4-fold, 1.35-fold, 1.3-fold, 1.25-fold, 1.2-fold, 1.15-fold, 1.1-fold, or 1.05-fold longer than that of an antibody comprising a wild-type Fc region. In certain embodiments, the increased half-life of the antibody is about 10,000-fold, 1,000-fold, 500-fold, 100-fold, 50-fold, 20-fold, 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4.5-fold, 4-fold, 3.5-fold, 3-fold, 2.5-fold, 2-fold, 1.95-fold, 1.9-fold, 1.85-fold, 1.8-fold, 1.75-fold, 1.7-fold, 1.65-fold, 1.6-fold, 1.55-fold, 1.50-fold, 1.45-fold, 1.4-fold, 1.35-fold, 1.3-fold, 1.25-fold, 1.2-fold, 1.15-fold, 1.1-fold, or 1.05-fold longer than dupilumab. In certain embodiments, the increased half-life of the antibody is about 2.5-fold, 2.4-fold, 2.3-fold, 2.2-fold, 2.1-fold, 2.0-fold, 1.9-fold, or 1.8-fold longer than dupilumab.

[0375] In certain embodiments, the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in reduced one or more of ADCC activity, ADCP activity, or CDC activity compared to an Fc region without the one or more substitutions.

[0376] In certain embodiments, the one or more amino acid substitutions are selected from the group consisting of S228P(SP), M252Y, S254T, T256E, T256D, T250Q, H285D, T307A, T307Q, T307R, T307W, L309D, Q411H, Q311V, A378V, E380A, M428L, N434A, N434S, N297A, D265A, L234A, L235A, and N434W. In certain embodiments, the one or more amino acid substitutions comprise a specific combination of amino acid substitutions selected from the group consisting of: M428L / N434S (LS); M252Y / S254T / T256E (YTE); T250Q / M428L; T307A / E380A / N434A; T256D / T307Q (DQ); T256D / T307W (DW); M252Y / T256D (YD); T307Q / Q311V / A378V (QVV); T256D / H285D / T307R / Q311V / A378V (DDRVV); L309D / Q311H / N434S (DHS); S228P / L235E (SP LE); L234A / L235A(LA), M428L / N434A, L234A / G237A(LAGA), L234A / L235A / G237A(LALAGA), L234A / L235A / P329G, D265A / YT E. LALA / YTE, LAGA / YTE, LALAGA / YTE, LALAPG / YTE, N297A / LS; D265A / LS; LALA / LS; LALAGA / LS; LALAPG / LS; N297A / DHS; D265A / DHS; LALA / DHS; LAGA / DHS; LALAGA / DHS ; LALAPG / DHS; SP / YTE; SPLE / YTE; SP / LS; SPLE / LS, SP / DHS; SPLE / DHS; N297A / LA; D265A / LA, LALA / LA, LAGA / LA, LALAGA / LA, LALAPG / LA, N297A / N434A; D265A / N434A;LALA / N434A, LAGA / N434A, LALAGA / N434A, LALAPG / N434A, N297A / N434W, D265A / N434W, LALA / N434W, LAGA / N434W, LALAGA / N434W, LAL APG / N434W, N297A / DQ, D265A / DQ, LALA / DQ, LAGA / DQ, LALAGA / DQ, LALAPG / DQ, N297A / DW, D265A / DW, LALA / DW, LAGA / DW, LALAGA / DW, LAL APG / DWN297A / YD, D265A / YD, LALA / YD, LAGA / YD, LALAGA / YD, LALAPG / YD, T307Q / Q311V / A378V(QVV), N297A / QVV, D265A / QVV, LALA / QV V, LAGA / QVV, LALAGA / QVV, LALAPG / QVV, DDRVV, N297A / DDRVV, D265A / DDRVV, LALA / DDRVV, LAGA / DDRVV, LALAGA / DDRVV and LALAPG / DDRVV. ;

[0377] In certain embodiments, the Fc region binds an Fcγ receptor selected from the group consisting of: FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb. In certain embodiments, the Fc region binds an Fcγ receptor with a higher affinity at pH 6.0 than an antibody comprising a wild-type Fc region.

[0378] Combine

[0379] The affinity of molecule X for its partner Y can be determined by the dissociation equilibrium constant (K D ) is represented. The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. The kinetic components that contribute to the dissociation equilibrium constant can be determined by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) techniques (e.g. ) or biolayer interferometry (e.g. ) to measure affinity.

[0380] With respect to the binding of an antibody to a target molecule, the terms "bind," "specifically binds," "binds specifically to," "has specificity for," "selectively binds," and "is selective for" a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from non-specific or non-selective interactions (e.g., with a non-target molecule). Specific binding can be measured, for example, by measuring binding to a target molecule (i.e., IL-4Rα) and comparing it to binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule. In such cases, specific binding is indicated if binding of the antibody to the target molecule is competitively inhibited by the control molecule. In some embodiments, the affinity of the anti-IL-4Rα antibody for the non-target molecule is less than about 50% of the affinity for IL-4Rα. In some embodiments, the affinity of the anti-IL-4Rα antibody for the non-target molecule is less than about 40% of the affinity for IL-4Rα. In some embodiments, the anti-IL-4Rα antibody has an affinity for a non-target molecule that is less than about 30% of its affinity for IL-4Rα. In some embodiments, the anti-IL-4Rα antibody has an affinity for a non-target molecule that is less than about 20% of its affinity for IL-4Rα. In some embodiments, the anti-IL-4Rα antibody has an affinity for a non-target molecule that is less than about 10% of its affinity for IL-4Rα. In some embodiments, the anti-IL-4Rα antibody has an affinity for a non-target molecule that is less than about 1% of its affinity for IL-4Rα. In some embodiments, the anti-IL-4Rα antibody has an affinity for a non-target molecule that is less than about 0.1% of its affinity for IL-4Rα.

[0381] When used herein in the context of two or more antibodies, the term "compete with" or "cross-compete with" indicates that the two or more antibodies compete for binding to an antigen (e.g., IL-4Rα). In one exemplary assay, IL-4Rα is coated on a surface and contacted with a first anti-IL-4Rα antibody, after which a second anti-IL-4Rα antibody is added. In another exemplary assay, a first anti-IL-4Rα antibody is coated on a surface and contacted with IL-4Rα, and then a second anti-IL-4Rα antibody is added. If the presence of the first anti-IL-4Rα antibody reduces the binding of the second anti-IL-4Rα antibody in either assay, the antibodies compete with each other. The term "compete with" also includes antibody combinations in which one antibody reduces the binding of another antibody, but in which no competition is observed when the antibodies are added in the opposite order. However, in some embodiments, the first antibody and the second antibody inhibit binding to each other, regardless of the order in which they are added. In some embodiments, an antibody reduces the binding of another antibody to its antigen by at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99%, as measured in a competitive binding assay. One skilled in the art can select the concentration of an antibody to use in a competition assay based on the antibody's affinity for IL-4 and the antibody's valency. The assays described in this definition are illustrative, and one skilled in the art can utilize any suitable assay to determine whether antibodies compete with each other. Suitable assays are described, for example, in Cox et al. "Immunoassay Methods", Assay Guidance Manual [Internet], updated December 24, 2014 (ncbi.nlm.nih.gov / books / NBK92434 / ; accessed September 29, 2015); Silman et al. (2001) Cytometry 44:30-37; and Finco et al. (2011) J. Pharm. Biomed. Anal. 54:351-358; each of which is incorporated by reference in its entirety.

[0382] A test antibody competes with a reference antibody if an excess of the test antibody (e.g., at least 2x, 5x, 10x, 20x, or 100x) inhibits or blocks, for example, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the binding of the reference antibody as measured in a competitive binding assay. Antibodies identified by competition assays (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to an adjacent epitope sufficiently close to the epitope bound by the reference antibody to cause steric hindrance. For example, a second competing antibody can be identified that competes with a first antibody described herein for binding to IL-4Rα. In some cases, the second antibody can block or inhibit, for example, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the binding of the first antibody as measured in a competitive binding assay. In certain cases, the second antibody may displace the first antibody by greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%.

[0383] In certain embodiments, the antibody binds to the IL-4Rα sequence shown in SEQ ID NOs: 237-240.

[0384] In certain embodiments, the antibody has a specific activity at less than or equal to about 1 x 10 -9 M, 2x 10 -9 M, 3x 10 -9 M, 4x 10 -9 M, 5x 10 -9 M, 6x 10 -9 M, 7x 10 -9 M, 8x 10 -9 M, 9x 10 - 9 M's K D Binds to the IL-4Rα sequence shown in SEQ ID NO: 237-240. In certain embodiments, the antibody has a specific activity at a concentration of less than or equal to about 1×10 -10 M's K D Binds to the IL-4Rα sequence shown in SEQ ID NO: 237-240. In certain embodiments, the antibody has a specific activity at a concentration of less than or equal to about 1×10 -9 M's K D Binds to human IL-4Rα.

[0385] In some embodiments, the antibodies provided herein have a specific affinity for the antibody at a level of less than or equal to about 0.01 x 10-8 M, 0.02x10 -8 M, 0.03x10 -8 M, 0.04x10 -8 M, 0.05x10 -8 M, 0.06x10 -8 M, 0.07x10 -8 M, 0.08x10 -8 M, 0.09x10 -8 M, 0.1x10 -8 M, 0.2x10 -8 M, 0.3x10 -8 M, 0.4x10 -8 M, 0.5x10 -8 M, 0.6x10 -8 M, 0.7x10 -8 M, 0.8x10 -8 M, 0.9x10 -8 M, 1x10 -8 M, 1.1x10 -8 M, 1.2x10 -8 M, 1.3x10 -8 M, 1.4x10 -8 M, 1.5x10 -8 M, 1.6x10 -8 M, 1.7x10 -8 M, 1.8x10 -8 M, 1.9x10 -8 M, 1.95x10 -8 M, 2x10 -8 M, 2.5x10 -8 M, 3x10 -8 M, 3.5x10 -8 M, 4x10 -8 M, 4.5x10 -8 M, 5x10 -8 M, 6x10 -8 M, 7x10 -8 M, 8x10 -8 M, 9x10 -8 M or 10x10 -8 M's K D In some embodiments, the antibodies provided herein bind to IL-4Rα at a level of less than or equal to about 0.01 x 10 -9 M, 0.02x10 -9 M, 0.03x10 -9 M, 0.04x10-9 M, 0.05x10 -9 M, 0.06x10 -9 M, 0.07x10 -9 M, 0.08x10 -9 M, 0.09x10 -9 M, 0.1x10 -9 M, 0.2x10 -9 M, 0.3x10 -9 M, 0.4x10 -9 M, 0.5x10 -9 M, 0.6x10 -9 M, 0.7x10 -9 M, 0.8x10 -9 M, 0.9x10 -9 M, 1x10 -9 M, 1.1x10 -9 M, 1.2x10 -9 M, 1.3x10 -9 M, 1.4x10 -9 M, 1.5x10 -9 M, 1.6x10 -9 M, 1.7x10 -9 M, 1.8x10 -9 M, 1.9x10 -9 M, 1.95x10 -9 M, 2x10 -9 M, 2.5x10 -9 M, 3x10 -9 M, 3.5x10 -9 M, 4x10 -9 M, 4.5x10 -9 M, 5x10 -9 M, 6x10 -9 M, 7x10 -9 M, 8x10 -9 M, 9x10 -9 M or 10x10 -9 M's K D Binds to IL-4Rα.

[0386] In some embodiments, the antibodies provided herein bind to the K of IL-4Rα. D In about 0.001-0.01x10 -8 M, 0.01-0.1x10 -8 M, 0.01-0.05x10 -8 M, 0.05-0.1x10 -8 M, 0.1-0.5x10 -8M, 0.5-1x10 -8 M, 0.25-0.75x10 -8 M, 0.25-0.5x10 -8 M, 0.5-0.75x10 -8 M, 0.75-1x10 -8 M, 0.75-2x10 -8 M, 1.1-1.2x10 -8 M, 1.2-1.3x10 -8 M, 1.3-1.4x10 -8 M, 1.4-1.5x 10 -8 M, 1.5-1.6x 10 -8 M, 1.6-1.7x 10 -8 M, 1.7-1.8x 10 -8 M, 1.8-1.9x 10 -8 M, 1.9-2x 10 -8 M, 1-2x 10 -8 M, 1-5x 10 -8 M, 2-7x 10 -8 M, 3-8x 10 - 8 M, 3-5x 10 -8 M, 4-6x 10 -8 M, 5-7x 10 -8 M, 6-8x 10 -8 M, 7-9x 10 -8 M, 7-10x 10 -8 M or 5-10x 10 -8 In some embodiments, the antibodies provided herein have a specific expression level of less than or equal to about 1 x 10 M, as measured by ELISA or any other suitable method known in the art. -8 M, or less than or equal to higher than 1x 10 -9 M's K D Binds to IL-4Rα.

[0387] In some embodiments, the antibodies provided herein have a specific activity at a concentration of less than or equal to about 10 x 10 -8 M, 9x 10 -8 M, 8x 10 -8 M, 7x 10 -8 M, 6x 10 -8 M, 5x 10 - 8M、4.5x 10 -8 M、4x 10 -8 M、3.5x 10 -8 M、3x 10 -8 M、2.5x 10 -8 M、2x 10 -8 M、1.98x 10 -8 M、1.95x10 -8 M、1.9x 10 -8 M、1.85x 10 -8 M、1.8x 10 -8 M、1.75x 10 -8 M、1.7x 10 -8 M、1.65x10 -8 M、1.6x 10 -8 M、1.55x 10 -8 M、1.50x 10 -8 M、1.45x 10 -8 M、1.4x 10 -8 M、1.3x 10 -8 M、1.2x 10 -8 M、1.1x 10 - 8 M、1x 10 -8 M、0.9x 10 -8 M、0.85x 10 -8 M、0.8x 10 -8 M、0.75x 10 -8 M、0.7x 10 -8 M、0.65x 10 -8 M、0.6x 10 -8 M、0.55x 10 -8 M、0.5x 10 -8 M、0.45x 10 -8 M、0.4x 10 -8 M、0.35x 10 -8 M、0.3x 10 -8 M、0.25x 10 -8 M、0.2x 10 -8 M、0.15x 10 -8 M、0.1x10 -8 M、0.05x 10 -8 M、0.01x 10 -8 M、0.005x 10 -8 M、0.001x10 -8M, 0.0005x 10 -8 M or 0.0001x 10 -8 M or smaller K D In some embodiments, the antibodies provided herein bind to IL-4Rα at a concentration between 5-3×10 -8 M, 4-2x 10 -8 M, 3-1x 10 -8 M, 1.9-1.8x 10 -8 M, 1.8-1.7x 10 -8 M, 1.7-1.6x 10 -8 M, 1.6-1.5x10 -8 M, 1.9-1.5x 10 -8 M, 1.5-1x 10 -8 M, 1-0.8x 10 -8 M, 1-0.5x 10 -8 M, 0.9-0.6x 10 -8 M, 0.7-0.4x 10 -8 M, 0.6-0.2x 10 -8 M, 0.5-0.3x 10 -8 M, 0.3-0.2x 10 -8 M, 0.2-0.1x 10 -8 M, 0.1-0.01x10 -8 M, 0.01-0.001x 10 -8 M or 0.001-0.0001x 10 -8 K between M D Binds to IL-4Rα.

[0388] In some embodiments, the antibodies provided herein bind to FcRn at pH 7.4 as compared to pH 6.0 with an affinity of about 10,000, 1,000, 500, 100, 50, 20, 10, 9, 8, 7, 6, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.95, 1.9, 1.85, 1.8, 1.75, 1.7, 1.65, 1.6, 1.55, 1.50, 1.45, 1.4, 1.3, 1.2, 1.1 or 1.05 (pH 7.4 / pH 6.0) as measured by ELISA or any other suitable method known in the art. In some embodiments, the antibodies provided herein bind to FcRn at pH 6.0 as compared to pH 7.4 with an affinity of about 1-0.8 x 10 -8M, 1-0.5x 10 -8 M, 0.9-0.6x 10 -8 M, 0.7-0.4x 10 -8 M, 0.6-0.2x 10 - 8 M, 0.5-0.3x 10 -8 M, 0.3-0.2x 10 -8 M, 0.2-0.1x 10 -8 M, 0.1-0.01x 10 -8 M, 0.01-0.001x 10 -8 M or 0.001-0.0001x 10 -8 M ratio (pH 6.0 / pH 7.4) binds to FcRn with an affinity.

[0389] Function

[0390] "Effect function" refers to a biological activity mediated by the Fc region of an antibody, which may vary depending on the antibody isotype. Examples of antibody effector functions include receptor ligand blocking, agonism, or antagonism, C1q binding to activate complement-dependent cytotoxicity (CDC), Fc receptor binding to activate antibody-dependent cellular cytotoxicity (ADCC), and antibody-dependent cellular phagocytosis (ADCP). In some embodiments, the effector function of the anti-IL-4Rα antibodies described herein is antagonism and blocks IL-4Rα from binding to IL-4 and / or IL-13.

[0391] Pharmaceutical composition

[0392] The present application provides compositions comprising antibodies, including pharmaceutical compositions comprising any one or more of the antibodies described herein and one or more pharmaceutically acceptable excipients. In some embodiments, the composition is sterile. The pharmaceutical composition generally comprises an effective amount of the antibody.

[0393] In addition to one or more of the antibodies disclosed herein, these compositions may contain pharmaceutically acceptable excipients, carriers, buffers, stabilizers or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material may depend on the route of administration, such as oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes.

[0394] Pharmaceutical compositions for oral administration may be in tablet, capsule, powder or liquid form. Tablets may include solid carriers such as gelatin or adjuvants. Liquid pharmaceutical compositions typically include liquid carriers such as water, petroleum, animal or vegetable oils, mineral oils or synthetic oils. They may include physiological saline solutions, dextrose or other sugar solutions or glycols such as ethylene glycol, propylene glycol or polyethylene glycol.

[0395] For intravenous, skin or subcutaneous injection or injection at the site of ailment, active ingredient will be in the form of an acceptable aqueous solution for parenteral administration, which does not contain pyrogens and has suitable pH, isotonicity and stability. Those skilled in the art can well use, for example, isotonic vehicles (such as sodium chloride injection, Ringer's injection (Ringer's Injection), lactated Ringer's injection) to prepare suitable solutions. Preservatives, stabilizers, buffers, antioxidants and / or other additives may be included as needed.

[0396] The anti-IL-4Rα antibody to be administered to an individual is preferably administered in a "therapeutically effective amount" or a "prophylactically effective amount" (as appropriate, but prophylaxis may be considered therapy), which is sufficient to show benefit to the individual. The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of the protein aggregation disease being treated. Treatment prescriptions, such as decisions on dosage, are within the purview of general practitioners and other physicians, and generally take into account the condition to be treated, the individual patient's condition, the site of delivery, the method of administration, and other factors known to the physician. Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (ed.), 1980.

[0397] Depending on the condition to be treated, the compositions may be administered alone or in combination with other therapies, either simultaneously or sequentially.

[0398] method

[0399] Preparation method

[0400] The antibodies described herein can be produced using recombinant methods and compositions, for example as described in U.S. Patent No. 4,816,567. In one embodiment, an isolated nucleic acid encoding an antibody described herein is provided. This nucleic acid may encode an amino acid sequence comprising the VL of the antibody and / or an amino acid sequence comprising the VH of the antibody (e.g., the light chain and / or heavy chain of the antibody) or an amino acid sequence comprising the VHH of a single domain antibody. In another embodiment, one or more vectors (e.g., expression vectors) comprising this nucleic acid are provided. In one embodiment, the nucleic acid is provided in a polycistronic vector. In another embodiment, a host cell comprising this nucleic acid is provided. In one embodiment, the host cell comprises (e.g., transformed with): (1) a vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antigen-binding polypeptide construct; or (2) a first vector comprising a nucleic acid encoding an amino acid sequence comprising the VL of the antigen-binding polypeptide construct, and a second vector comprising a nucleic acid encoding an amino acid sequence comprising the VH of the antigen-binding polypeptide construct. In one embodiment, the host cell is a eukaryotic cell, such as a Chinese hamster ovary (CHO) cell or a human embryonic kidney (HEK) cell or a lymphocyte (e.g., a Y0, NS0, Sp20 cell). In one embodiment, a method for manufacturing an antibody is provided, wherein the method includes culturing a host cell comprising a nucleic acid encoding an antibody as provided above under conditions suitable for expressing the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0401] For recombinant production of antibodies, nucleic acids encoding antibodies, such as those described above, are isolated and inserted into one or more vectors for further cloning and / or expression in a host cell. Such nucleic acids can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that specifically bind to the genes encoding the heavy and light chains of the antibody).

[0402] When the antibody or variant thereof is recombinantly produced by a host cell, in certain embodiments, the protein is present at about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% or less of the dry weight of the cells. When the antibody or variant thereof is recombinantly produced by a host cell, in certain embodiments, the protein is present in the culture medium at about 5 g / L, about 4 g / L, about 3 g / L, about 2 g / L, about 1 g / L, about 750 mg / L, about 500 mg / L, about 250 mg / L, about 100 mg / L, about 50 mg / L, about 10 mg / L, or about 1 mg / L or less of the dry weight of the cells. In certain embodiments, "substantially purified" antibodies produced by the methods described herein have a purity level of at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, specifically, a purity level of at least about 75%, 80%, 85%, and more specifically, a purity level of at least about 90%, at least about 95%, at least about 99% or greater, as determined by appropriate methods such as SDS / PAGE analysis, RP-HPLC, SEC, and capillary electrophoresis.

[0403] Suitable host cells for cloning or expressing antibody-encoding vectors include prokaryotic or eukaryotic cells described herein.

[0404] Recombinant host cells or host cells are cells that include exogenous polynucleotides, regardless of the method used for insertion (e.g., direct uptake, transduction, f-coupling, or other methods known in the art for producing recombinant host cells). The exogenous polynucleotides can be maintained as non-integrating vectors (e.g., plasmids) or can be integrated into the host genome. Host cells can include CHO, CHO derivatives, NSO, Sp2O, CV-1, VERO-76, HeLa, HepG2, Per.C6, or BHK.

[0405] For example, antibodies can be produced in bacteria, especially when glycosylation and Fc effector functions are not required. For expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Patents 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (BKCLo ed., Humana Press, Totowa, NJ, 2003), pp. 245-254, which describes expression of antibody fragments in E. coli. After expression, the antibodies can be separated from the bacterial cell paste in a soluble fraction and can be further purified.

[0406] In addition to prokaryotes, eukaryotic microorganisms such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungal and yeast strains whose glycosylation pathways have been "humanized" so that the antibodies produced have a partially or fully human glycosylation pattern. See Gerngross (2004) Nat. Biotech. 22: 1409-1414 and Li et al. (2006) Nat. Biotech. 24: 210-215.

[0407] Suitable host cells for expressing glycosylated antibodies also come from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Several baculovirus strains have been identified that can be used in conjunction with insect cells, particularly for transfecting Spodoptera frugiperda cells.

[0408] Plant cell cultures can also be used as hosts. See, for example, U.S. Patent Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing plant cell cultures for producing antibodies in transgenic plants). TM technology).

[0409] Vertebrate cells can also be used as hosts. For example, mammalian cell lines adapted to growth in suspension can be used. Other examples of suitable mammalian host cell lines are monkey kidney CV1 strain transformed by SV40 (COS-7); human embryonic kidney strain (293 or 293 cells, as described, for example, in Graham et al. (1977) J. Gen Virol. 36:59); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells, as described, for example, in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK); buffalo rat liver cells (BRL 3A); human lung cells (W138); human hepatocytes (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, for example, in Mather et al. Annals of . Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other suitable mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al. (1980) Proc. Natl. Acad. Sci. USA 77:4216); and myeloma cell lines, such as Y0, NS0, and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKCLo ed., Humana Press, Totowa, NJ), pp. 255-268 (2003).

[0410] In one embodiment, the antibodies described herein are produced in stable mammalian cells by a method comprising: transfecting at least one stable mammalian cell with nucleic acids encoding the antibodies at a predetermined ratio; and expressing the nucleic acids in the at least one mammalian cell. In some embodiments, the predetermined ratio of nucleic acids is determined in a transient transfection experiment to determine the relative ratio of input nucleic acids that produces the highest percentage of antibody in the expression product.

[0411] In some embodiments are methods of producing an antibody in a stable mammalian cell as described herein, wherein at least one expression product of the stable mammalian cell comprises a greater percentage of the desired glycosylated antibody compared to a monomeric heavy or light chain polypeptide or other antibody.

[0412] In some embodiments, a method for producing a glycosylated antibody in a stable mammalian cell as described herein comprises identifying and purifying the desired glycosylated antibody. In some embodiments, the identification is by one or both of liquid chromatography and mass spectrometry.

[0413] If necessary, the antibody can be purified or isolated after expression. Proteins can be isolated or purified in a variety of ways known to those skilled in the art. Standard purification methods include chromatographic techniques, including ion exchange, hydrophobic interaction, affinity, size determination or gel filtration and reverse phase, which are performed at atmospheric pressure or under high pressure using systems such as FPLC and HPLC. Purification methods also include electrophoresis, immunoassays, precipitation, dialysis and chromatofocusing techniques. Ultrafiltration and diafiltration techniques combined with protein concentration are also used. As is well known in the art, a variety of natural proteins bind to Fc and antibodies, and these proteins can be used in the present invention to purify antibodies. For example, bacterial proteins A and G bind to the Fc region. Similarly, bacterial protein L binds to the Fab region of some antibodies. Purification can often be achieved by specific fusion partners. For example, glutathione resins (if GST fusions are used), Ni +2 The antibody is purified by affinity chromatography (if a His tag is used) or by immobilizing an anti-flag antibody (if a flag tag is used). For general guidance on suitable purification techniques, see, for example, Protein Purification: Principles and Practice, 3rd Edition, Scopes, Springer-Verlag, NY, 1994, which is incorporated by reference in its entirety. The degree of purification required will vary depending on the use of the antibody. In some cases, no purification is required.

[0414] In certain embodiments, antibodies are purified using anion exchange chromatography, including but not limited to chromatography on Q-Sepharose, DEAE Sepharose, Poros HQ, Poros DEAF, Toyopearl Q, Toyopearl QAE, Toyopearl DEAE, Resource / Source Q and DEAE, Fractogel Q and DEAE columns.

[0415] In certain embodiments, the proteins described herein are purified using cation exchange chromatography, including but not limited to SP-Sepharose, CM Sepharose, Poros HS, Poros CM, Toyopearl SP, Toyopearl CM, Resource / Source S and CM, Fractogel S and CM columns, and equivalents and analogs thereof.

[0416] In addition, the antibodies described herein can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, WH Freeman & Co., NY and Hunkapiller et al. (1984) Nature, 310: 105-111). For example, polypeptides corresponding to polypeptide fragments can be synthesized using a peptide synthesizer. In addition, if necessary, non-classical amino acids or chemical amino acid analogs can be introduced as substitutions or additions to the polypeptide sequence. In general, non-classical amino acids include, but are not limited to, D-isomers of common amino acids, 2,4-diaminobutyric acid, α-aminoisobutyric acid, 4-aminobutyric acid, Abu, 2-aminobutyric acid, g-Abu, e-Ahx, 6-aminohexanoic acid, Aib, 2-aminoisobutyric acid, 3-aminopropionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteine, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, alanine, fluoroamino acids, designer amino acids such as methyl amino acids, C-methyl amino acids, N-methyl amino acids, and amino acid analogs. In addition, the amino acids can be D (dextrorotatory) or L (levorotatory).

[0417] How to use

[0418] In one aspect, the application provides methods of contacting IL-4Rα with an anti-IL-4Rα antibody (eg, a human or humanized antibody) that results in inhibition of IL-4 binding to IL-4 receptor α expressed on a cell.

[0419] In one aspect, the present application provides a method of using the isolated anti-IL-4Rα antibodies described herein to treat a condition or disease in a subject. In certain aspects, described herein is a method of using an anti-IL-4Rα antibody to treat a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount of an anti-IL-4Rα antibody or a pharmaceutical composition comprising an anti-IL-4Rα antibody described herein. In certain embodiments, the present application provides a method of treating a condition or disease associated with elevated IL-4 and / or IgE levels in a subject.

[0420] In certain aspects, described herein are methods for treating a disorder associated with IL-4, IL-13, and / or IL-4Rα activity, comprising administering to a mammalian subject a therapeutically effective amount of an isolated anti-IL-4Rα antibody or a pharmaceutical composition comprising an isolated anti-IL-4Rα antibody described herein.

[0421] In certain aspects, the antibodies and antibody fragments disclosed herein are useful for treating diseases and conditions that are improved, inhibited, or ameliorated by reducing the activity of IL-4, IL-13, and / or IL-4Rα. These conditions include conditions characterized by abnormal or overexpression of IL-4 and / or IL-13, or abnormal host responses to IL-4 and / or IL-13. IL-4 and IL-13-related conditions treated by the antibodies or antibody fragments disclosed herein include inflammatory conditions or diseases. Non-limiting examples of disorders that can be treated by the antibodies or antibody fragments of the present disclosure include atopic dermatitis (AD), asthma (mild, moderate or severe), chronic sinusitis with nasal polyps, chronic sinusitis without nasal polyps (CRSsNP), eosinophilic esophagitis (EoE), an eosinophilic gastrointestinal disorder or disease (ENID) selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC) and eosinophilic gastroenteritis (EGE); Chargar-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA), prurigo nodularis (PN), chronic spontaneous urticaria (CSU), chronic pruritus of unknown origin (CPUO), bullous pemphigoid (BP), cold-induced urticaria (ColdU), allergic fungal sinusitis (AFRS), allergic bronchopulmonary aspergillosis (ABPA), chronic obstructive pulmonary disease (COPD), inflammatory bowel disease such as Crohn's disease or ulcerative colitis, lupus, and rheumatoid arthritis.

[0422] In certain aspects, described herein are methods for treating an inflammatory condition or disease in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein. In certain embodiments, the inflammatory condition or disease is atopic dermatitis. In certain embodiments, the inflammatory condition or disease is asthma. In certain embodiments, the inflammatory condition or disease is nasal polyps.

[0423] In certain aspects, described herein are methods for treating a pathology associated with elevated levels of IL-4 and / or IL-13 in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0424] In certain aspects, described herein are methods of reducing the biological activity of IL-4, IL13, and / or IL-4Rα in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0425] In certain aspects, described herein are methods of preventing an inflammatory disorder or disease in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0426] Application method

[0427] In some embodiments, the methods provided herein are suitable for treating a disease or condition in a subject. In one embodiment, the subject is a human and the antibody is an anti-IL-4Rα antibody described herein.

[0428] In some embodiments, the antibody is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implant, by inhalation, intrathecally, intracerebroventricularly, or intranasally. An effective amount of the anti-IL-4Rα antibody can be administered for the treatment of a disease or condition. The appropriate dose of the anti-IL-4Rα antibody can be determined based on the type of disease or condition being treated, the type of anti-IL-4Rα antibody, the severity and course of the disease or condition, the individual's clinical condition, the individual's clinical history and response to treatment, and the judgment of the attending physician.

[0429] In certain embodiments, the anti-IL-4Rα antibody is administered every six weeks or every two months.

[0430] In some embodiments, the antibodies provided herein are administered together with at least one additional therapeutic agent. Any suitable additional therapeutic agent or immunotherapeutic agent can be administered together with the antibodies provided herein. Additional therapeutic agents include agents for treating or preventing diseases or conditions associated with elevated IL-4, IL-13, and / or IgE levels, such as, but not limited to, inflammatory diseases or conditions.

[0431] The additional therapeutic agent can be administered by any suitable means. In some embodiments, the antibody provided herein and the additional therapeutic agent are included in the same pharmaceutical composition. In some embodiments, the antibody provided herein and the additional therapeutic agent are included in different pharmaceutical compositions.

[0432] In the embodiment that the antibody that this paper provided and additional therapeutic agent are included in different pharmaceutical compositions, can before, simultaneously and / or after using additional therapeutic agent, carry out antibody administration.In some embodiments, the antibody that this paper provided and the use of additional therapeutic agent are carried out within about one month of each other.In some embodiments, the antibody that this paper provided and the use of additional therapeutic agent are carried out within about one week of each other.In some embodiments, the antibody that this paper provided and the use of additional therapeutic agent are carried out within about one day of each other.In some embodiments, the antibody that this paper provided and the use of additional therapeutic agent are carried out within about twelve hours of each other.In some embodiments, the antibody that this paper provided and the use of additional therapeutic agent are carried out within about one hour of each other.

[0433] Medicine boxes and products

[0434] The present application provides a kit comprising any one or more of the antibody compositions described herein and instructions for use. In some embodiments, the kit further comprises a component selected from any one of the following: a secondary antibody, a reagent for immunohistochemical analysis, a pharmaceutically acceptable excipient, a drug insert and an instruction manual, and any combination thereof. In a specific embodiment, the kit comprises a pharmaceutical composition comprising any one or more of the antibody compositions described herein and one or more pharmaceutically acceptable excipients.

[0435] The present application also provides an article of manufacture comprising any of the antibody compositions or kits described herein. Examples of articles of manufacture include vials (including sealed vials).

[0436] Example

[0437] Below are examples for carrying out specific embodiments of the present invention. The examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way. Every effort has been made to ensure the accuracy of the numerical values ​​(e.g., amounts, temperatures, etc.) used, but of course some experimental errors and deviations should be allowed.

[0438] Unless otherwise indicated, the practice of the present invention will employ conventional methods of protein chemistry, biochemistry, recombinant DNA technology, and pharmacology within the skill of the art. Such techniques are fully explained in the literature. See, for example, T. C. Reighton, Proteins: Structures and Molecular Properties (W. H. Freeman and Company, 1993); A. L. Lehninger, Biochemistry (Worth Publishers, Inc., current edition); Sambrook et al., Molecular Cloning: A Laboratory Manual (2nd edition, 1989); Methods In Enzymology (S. Colowick and N. Kaplan, eds., Academic Press, Inc.); Remington's Pharmaceutical Sciences, 18th edition (Easton, Pennsylvania: Mack Publishing Company, 1990); Carey and Sundberg Advanced Organic Chemistry, 3rd edition (Plenum Press) Volumes A and B (1992).

[0439] method

[0440] Gene synthesis and plasmid construction

[0441] The coding sequences of the antibody HC and LC were generated by DNA synthesis and PCR, and then subcloned into a pTT5-based plasmid for protein expression in mammalian cell systems. The gene sequence in the expression vector was confirmed by DNA sequencing.

[0442] Purification of antibody constructs

[0443] Protein purification was performed by affinity chromatography and ion exchange chromatography using an AKTA pure instrument (GE Lifesciences). Conditioned culture medium expressing the target antibody was harvested by centrifugation at 4000 rpm for 50 min and filtered with a 0.22 μm filter. The harvested supernatant was loaded onto a Mabselect TM SuRe TM(GE Healthcare) column. After washing the column with buffer A (PBS, pH 7.4), the protein was eluted with buffer B (1 M glycine, pH 2.7) and immediately neutralized with 1 / 10 volume of buffer D (1 M sodium citrate, pH 6.0). The affinity purified antibody buffer was then exchanged into 20 mM sodium acetate pH 5.5.

[0444] Measuring Antibody IL-4 Binding Kinetics Using Surface Plasmon Resonance

[0445] The protein G sensor chip (Cytiva, catalog number 29179315) was used. 8KSPR system (GE Healthcare) was used to determine binding kinetic rates and affinity constants at 25°C in HBS-EP+ (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% surfactant P20). After a stabilization period in the electrophoresis buffer, anti-IL13 mAb constructs (diluted to 1 μg / mL) were captured onto flow cell 2 (active) for 60 seconds at a flow rate of 10 μL / min. Recombinant human IL-4 protein, His tag (Acro, catalog number IL3-H52H4) was prepared at concentrations of 0, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, and 0 nM and injected onto flow cell 1 (reference) and flow cell 2 (active) for 180 seconds at a flow rate of 30 μL / min. Recombinant cynomolgus macaque IL-4 protein His tag (SINO BIOLOGICAL, catalog number 11057-C07H) is prepared with a concentration of 0, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25 and 0nM and injected into flow cell 1 (reference) and flow cell 2 (activity) for 180 seconds at a flow rate of 30 μL / min. The capture surface is regenerated by injecting glycine pH 1.5 for 30 seconds at a flow rate of 30 μL / min and the sample is injected into the freshly captured mAb in a multi-cycle manner. Data were processed and analyzed using InsightEvaluation software, version 2.0.15.12933 (GE Healthcare). The response in flow cell 1 (reference) was subtracted from the response in flow cell 2 (active). The responses of two buffer blank injections were then subtracted from the reference-subtracted data (2-1), yielding doubly referenced data, which were fitted to a 1:1 binding model to determine the apparent association (ka) and dissociation rate constants (kd). The ratio of these provides the apparent equilibrium dissociation constant or affinity constant (KD = kd / ka).

[0446] Example 1: Affinity maturation of anti-IL-4Rα antibodies

[0447] Dupilumab was used as the parental antibody for further CDR diversification to identify clones with improved potency, manufacturability, and pharmacokinetics.

[0448] To diversify the CDRs while at least maintaining the potency of dupilumab, various mutations were made in all CDRs of the heavy and light chains. Site-directed PCR mutagenesis was used to generate different libraries for each CDR in the heavy and light chains, resulting in six unique libraries with an average of 2-3 amino acid substitutions per chain. Individual mutants were displayed as Fabs in a phage display system, and within each library, mutants were selected through two rounds of selection. The first round of selection consisted of 1 nM biotinylated hIL-4Rα and the output of that round was then divided into four different secondary rounds of parallel selection consisting of: A) 1 nM biotinylated hIL-4Rα while also washing in buffer containing 100 nM unlabeled hIL-4Rα for 4 hours at room temperature, B) 0.1 nM biotinylated hIL-4Rα while also washing in buffer containing 10 nM unlabeled hIL-4Rα for 4 hours at room temperature, C) 50 nM biotinylated cyIL-4Rα without washing and D) 5 nM biotinylated cyIL-4Rα without washing.

[0449] The output mutant clones from each set of secondary selection were analyzed as a proxy for binding affinity using periplasmic extracts of each mutant clone, using surface plasmon resonance (SPR) and comparison with dupilumab, and using bioinformatic analysis of the sequences to identify enrichment patterns. off Mutants that exhibited no loss or improved binding relative to dupilumab and that were enriched across the selected population were pooled into a single library containing all mutants in both the heavy and light chains. Fabs containing random combinatorial mixtures of these individual CDR mutants were screened again in a phage display system with multiple combinations of selection strategies summarized in Table 5.

[0450] Table 5

[0451]

[0452] *Output from round 2 selection in this strategy was selected in a third round consisting of 5 nM biotinylated cyIL-4Rα with a 16 h RT wash and 500 nM unlabeled cyIL-4Rα.

[0453] Example 2: Determination of antibody affinity for IL-4Rα

[0454] The binding affinity (KD) of the antibodies to human and cynomolgus macaque IL-4Rα was determined using Carterra LSA via surface plasmon resonance (SPR). Briefly, a HC30M sensor chip was previously functionalized with a polyclonal mixture of goat anti-human Fc antibodies and used to capture purified antibodies at levels between 100-1000 RLU. Subsequently, antigens at concentrations ranging from 200 nM to 0.13 nM were injected onto the surface at a rate of 2 mL / min. The chip was regenerated between different concentrations of antigen using 200 mM phosphoric acid and the antibodies were captured again as previously described. The KD was then derived using a 1:1 Langmuir binding model fitted to the 1:1 Langmuir binding model. D The association and dissociation rate constants were determined using Carterra's kinetics software. The results are summarized in Table 6.

[0455] Most antibodies bound to human IL-4Rα with low picomolar affinities comparable to dupilumab. In addition, several antibodies exhibited improved cross-reactivity with cynomolgus macaque IL-4Rα, some with low nanomolar affinities.

[0456] Table 6

[0457]

[0458]

[0459]

[0460] WS weak signal, below Carterra sensitivity.

[0461] For the relative value of dupilumab, <1.00 indicates tighter affinity.

[0462] Example 3: Improving the affinity of antibodies for FcRn at pH 6.0

[0463] The binding affinity (K) of the antibodies to human and cynomolgus macaque FcRn at pH 6.0 D ) is a reliable correlate of in vivo half-life. 8K determination of the K of purified antibodies to FcRn DBriefly, purified antibodies normalized to 0.5 mg / mL were captured using a protein G functionalized SPR chip with a flow rate of 10 μL / min for 60 s. A paired channel with buffer only was used as a reference. Subsequently, human or cynomolgus macaque FcRn at concentrations ranging from 25 nM to 0.39 nM was injected over the surface with the captured purified antibodies as well as the reference channel. Regeneration of the chip between different concentrations of IL-13 was performed with 10 mM glycine HCl, pH 1.5, and antibodies were captured again as previously described. Affinity measurements as described were performed at both pH 7.4 and pH 6.0. K was then derived using a 1:1 Langmuir binding model. D Value Insight Evaluation software was used to determine association and dissociation rate constants. All of the modified antibodies were expected to show enhanced binding to FcRn at pH 6.0 relative to dupilumab, a strong indicator that these antibodies will have increased in vivo half-lives. This was further confirmed in pharmacokinetic studies in cynomolgus monkeys.

[0464] Example 4: Inhibition of IL-13 and IL-4 binding to hIL-13Rα / hIL-4Rα

[0465] The functional blocking of antibodies against this binding interaction was evaluated using the combination of IL-13 and IL-4 with cells overexpressing hIL-13Rα / hIL-4Rα. Briefly, HEK293 cells previously transduced to stably express both hIL-13Rα and hIL-4Rα were cultured and harvested. Cells were seeded with 200,000 cells (100 μL) per well. The cells were washed and the supernatant was discarded. 100 μL of a mixture of biotinylated hIL-13 or biotinylated hIL-4 and purified antibodies (1: 1, by volume) previously prepared and incubated for 1 hour was added to resuspend the cells, resulting in a final concentration of 0.05 μg / mL hIL-13 or 0.04 μg / mL hIL-4 and 0-100 nM purified antibodies. The cells were stained in this mixture for 1 hour at 4°C. The cells were then washed and stained with 100 μL of Alexa Fluor 488-conjugated streptavidin diluted 1:1000 to detect the binding of biotinylated hIL-13 or biotinylated hIL-4 on the cell surface. The cells were incubated at 4°C for 1 hour, protected from light. The cells were then washed and the MFI of the cells in each well was recorded by FACS using a BD FACSCanto II. The data were then analyzed using GraphPad Prism. The IC50 value was determined as the antibody concentration required to inhibit 50% of the maximum MFI detected on the surface of biotinylated hIL-13 or biotinylated hIL-4 in the presence of 0.05 μg / mL hIL-13 or 0.04 μg / mL hIL-4 alone. The results are summarized in Tables 7, 8, Figure 1A and Figure 1B middle.

[0466] Table 7

[0467]

[0468]

[0469]

[0470] All values ​​are relative to dupilumab. <1.000 is more potent.

[0471] Certain constructs shown in Table 8 were modified to replace the IgG4 YTE heavy chain constant region with the IgG1 YTE LALA heavy chain constant region (SEQ ID NO: 321). These antibodies were tested for their ability to inhibit IL-13 and IL-4 binding to hIL-13Rα / hIL-4Rα compared to dupilumab. The results are summarized in Table 8. Values ​​reflect absolute IC 50 Measurement.

[0472] Table 8

[0473]

[0474]

[0475] Example 5: Inhibition of IL-13-induced and IL-4-induced STAT6 phosphorylation in HT-29 cells

[0476] The functional activity of the biological activity of antibody blocking IL-13 induction and IL-4 induction is evaluated using the inhibition of STAT6 phosphorylation in HT-29 cells. In short, HT-29 cells are starved overnight in RMPI 1640+0.1% FBS. Cells are collected and inoculated with 50,000 cells / well (100 μL). Meanwhile, a mixture of 100 μL hIL-13 or hIL-4 and purified antibodies (1: 1, by volume) is added to the same well to produce a final concentration of 10 ng / mL hIL-13 or 5 ng / mL hIL-4 and 0-50 nM purified antibodies. Cells are incubated at 37°C for 1 hour, and subsequently fixed, infiltrated, and stained with the anti-pSTAT6 antibody conjugated to PE. By FACS, the MFI of cells in each well is recorded using BD FACSCanto II, and subsequent data are analyzed using GraphPad Prism. IC50 values ​​were determined as the antibody concentration required to inhibit 50% of the maximum MFI of pSTAT6 detected in the presence of 10 ng / mL hIL-13 or 5 ng / mL hIL-4 alone. Figure 2A and Figure 2B middle.

[0477] As shown in Table 9, clones were identified that exhibited up to approximately 2-fold more potent inhibition of IL-13 and / or IL-4-induced STAT6 phosphorylation compared to dupilumab (e.g., Construct 15, Construct 18, Construct 21, Construct 23, Construct 24, Construct 27, and Construct 28).

[0478] Table 9

[0479]

[0480]

[0481]

[0482] All values ​​are relative to dupilumab. <1.000 is more potent.

[0483] The constructs shown in Table 10 were modified to replace the IgG4YTE heavy chain constant region with the IgG1YTE LALA constant region (SEQ ID NO: 321). These constructs were tested for their ability to inhibit IL-13 and / or IL-4-induced STAT6 phosphorylation compared to dupilumab. The results are summarized in Table 10. Values ​​reflect absolute IC 50 Measurement.

[0484] Table 10

[0485]

[0486] Example 6: Inhibition of IL-13-induced and IL-4-induced TARC release from A549 cells

[0487] The functional activity of the biological activity that antibody blocks IL-13 induction and IL-4 induction is evaluated using the inhibition of A549 cell secretion TARC.In brief, A549 cells are seeded in DMEM+10% FBS with 20,000 cells (100 μ L) and cultured overnight at 37 DEG C.On the second day, cell culture medium is discarded, and cells are washed gently with fresh culture medium.A mixture of 150 μ L hIL-13, purified antibodies and hTNFa (1:1:1, by volume) is added into the wells to produce the ultimate concentration of 20ng / mL hIL-13, 0-100nM purified antibodies and 200ng / mL hTNFa, or 1.5ng / mL hIL-4, 0-100nM purified antibodies and 50ng / mLTNFa.Cells are incubated in this mixture for 20-24 hours at 37 DEG C. After incubation, culture supernatants were collected and analyzed for the presence of TARC using a commercially available TARC ELISA kit (R&D Systems) according to the manufacturer's instructions. The TARC concentration determined in each well was analyzed using GraphPad Prism. The IC50 value was determined as the antibody concentration required to inhibit 50% of the maximum TARC concentration detected in the presence of 20 ng / mL hIL-13 alone and 200 ng / mL hTNFα or 1.5 ng / mL hIL-4 and 50 ng / mL hTNFα. The results are summarized in Tables 11, 12, and 13. Figure 3A and Figure 3B middle.

[0488] As shown in Table 11, clones were identified that exhibited up to approximately 3-fold more potent inhibition of IL-13 and / or IL-4-induced TARC compared to dupilumab (eg, Construct 2 and Construct 15).

[0489] Table 11

[0490]

[0491]

[0492] All values ​​are relative to dupilumab. <1.000 is more potent.

[0493] The constructs shown in Table 12 were modified to replace the IgG4 YTE heavy chain constant region with the IgG1 YTE LALA heavy chain constant region (SEQ ID NO: 321). These constructs were tested for their ability to inhibit IL-13 and / or IL-4-induced TARC compared to dupilumab. The results are summarized in Table 12. Values ​​reflect absolute IC 50 Measurement.

[0494] Table 12

[0495]

[0496] Example 7: Inhibition of IL-13-induced and IL-4-induced proliferation of TF-1 cells

[0497] The functional activity of the antibody blocking IL-13 induction and IL-4 induction biological activity was evaluated using the proliferation of TF-1 cells or its inhibition. In short, TF-1 cells were harvested and starved for 4 hours in RPMI1640+10% FBS without additional cytokines. During this period, a mixture of hIL-13 or hIL-4 and purified antibodies was prepared (1:1, by volume), and 50 μL was added to each well. After starvation, TF-1 cells were harvested again and inoculated with 15,000 cells (50 μL) per well to produce a final concentration of 4 ng / mL hIL-13 or 0.5 ng / mL hIL-4 and 0-5 nM purified antibodies. The cells were then incubated at 37°C for 72 hours, and the proliferation of cells was quantified using CellTiter-Glo (Promega) according to the manufacturer's instructions. Luminescence was recorded by a SpectraMaxM5 multi-mode plate reader, and data were analyzed using GraphPad Prism. IC50 values ​​were determined as the antibody concentration required to elicit 50% of the maximum luminescence detected when TF-1 cells were incubated and cultured with 4 ng / mL hIL-13 or 0.5 ng / mL hIL-4 alone. The results are summarized in Tables 13, 14, Figure 4A and Figure 4B middle.

[0498] Table 13

[0499]

[0500]

[0501]

[0502] All values ​​are relative to dupilumab. <1.000 is more potent.

[0503] The constructs shown in Table 14 were modified to replace the IgG4 YTE heavy chain constant region with the IgG1 YTE LALA heavy chain constant region (SEQ ID NO: 321). These constructs were tested for their ability to inhibit IL-13-induced and / or IL-4-induced TF-1 proliferation compared to dupilumab. The results are summarized in Table 14. Values ​​reflect absolute IC 50 Measurement.

[0504] Table 14

[0505]

[0506] Example 8: Improving the developability of anti-IL-4Rα antibodies

[0507] When the antibody itself has improved biophysical characteristics, the manufacture and clinical performance of the antibody are generally improved. In order to assess some of these features, the hydrophobic properties and nonspecificity of the purified antibodies are analyzed. In brief, for hydrophobic properties, 10 μg of purified antibodies are injected into hydrophobic interaction chromatography (HIC) Butyl 1.7 μm posts in the mobile phase of sodium phosphate, so that the antibody is captured to the post, then the ammonium sulfate retention time elution antibody is used as a direct proportional measurement of hydrophobicity, that is, the hydrophobicity feature of the shorter indication antibody of retention time is lower.

[0508] ELISA is used to assess nonspecificity. In brief, ELISA plates were coated with 0.15% baculovirus particle (BVP) suspension and incubated overnight at 4°C. Plates were blocked with 1% BSA at 37°C for 2 hours. 200nM purified antibodies were added and incubated at 37°C for 1 hour. Goat anti-human IgG Fc antibodies conjugated to HRP were used to detect the binding of the purified antibodies, which were incubated at 37°C for 0.5 hours. Between all steps, the plates were washed multiple times in PBS-T. The plates were developed using substrate TMB over a 15-minute time course, with the reaction stopped by adding 1N HCl. The absorbance at 450nm was recorded by a SpectraMax Plus 384 microplate reader and the data were analyzed using GraphPad Prism. The absorbance at 450nm for each antibody was used as a nonspecific proportional measurement, i.e., the smaller the absorbance, the lower the nonspecific binding expected for the antibody. The results are summarized in Table 15.

[0509] As shown in Table 15 below, almost all antibodies showed lower HIC and BVP scores than dupilumab.

[0510] Table 15

[0511]

[0512]

[0513]

[0514] NT: Not tested

[0515] Example 9. Improving the solubility and viscosity of anti-IL-4Rα antibodies

[0516] The purified antibodies were analyzed for maximum solubility and corresponding viscosity. Briefly, for maximum solubility, the purified antibodies were formulated into 20 mM histidine, 4% sucrose, 0.04% PS80, and buffers at pH X, Y, and Z, and concentrated. The concentration of the antibody at this point was evaluated and determined as the maximum solubility of the antibody at that pH. Subsequently, the viscosity of the antibody at that concentration and pH was determined. Several antibodies exhibited improved solubility greater than 150 mg / mL and greater viscosity relative to dupilumab, which allowed these antibodies to be delivered at higher doses per administration and reduced the frequency of injections to patients.

[0517] Example 10. Determination of antibody affinity for IL-4Rα

[0518] The binding affinity (KD) of the exemplary antibodies to human IL-4Rα was determined using a kinetic exclusion assay (KinExA). Briefly, antibodies and recombinant human IL-4Rα were equilibrated in solution. For each antibody, equilibrium samples were performed with fixed concentrations ranging from 2.27 pM to 100 pM of antibody and titration of recombinant human IL-4Rα produced at 2-fold dilutions starting from a starting concentration ranging from 9.78 pM to 1.00 nM, for a total of 11 to 13 concentrations. The samples were incubated for a period ranging from 3.5 hours to 340 hours to fully reach equilibrium. Once the samples reached equilibrium, azlactone beads previously coated with recombinant human IL-4Rα were used to capture any free antibody from the equilibrium solution. The capture antibody was then detected with goat anti-human IgG labeled with Alexa Fluor 647. A KinExA 4000 instrument was used to capture the fluorescent signal and convert it into a voltage signal that is proportional to the amount of free antibody in all equilibrium samples, thereby allowing the apparent affinity of the antibody to be determined. The results are summarized in Table 16.

[0519] Table 16

[0520]

[0521] Most of the exemplary antibodies bound to human IL-4Rα with femtomolar affinity, compared to dupilumab, which bound with low picomolar affinity.

[0522] Example 11. Pharmacokinetic Analysis of Anti-IL-4Rα Antibodies

[0523] In vivo pharmacokinetic (PK) studies were performed to evaluate the half-life extension of the antibodies relative to dupilumab. Cynomolgus macaques (Macaca fascicularis) were used for the study, with all matched SQ / IV groups being male and weighing between 2.52 and 3.86 kg. On day 0, animals were administered dupilumab, exemplary antibody construct 13 (mAb422), or exemplary antibody construct 38 (mAb471), where the IgG4 YTE constant heavy chain region of constructs 13 and 38 was replaced with the IgG1 YTELALA constant region (SEQ ID NO: 321), at a dose of 1, 5, or 25 mg / kg (as shown in Table 17), by intravenous (IV) bolus or subcutaneous (SQ) injection, and serum samples were obtained regularly throughout the study.

[0524] Table 17

[0525]

[0526] All SC group values ​​were adjusted for bioavailability

[0527] PK parameters were determined for serum samples of cynomolgus macaques up to day 91. PK analysis showed that the half-lives of construct 13 and construct 38 were 17.62 days and 25.60 days, respectively, compared to 10.88 days for dupilumab (25 mg / kg, IV). The results are summarized in Table 17. PK analysis showed that the exemplary antibodies construct 13 (mAb422) and construct 38 (mAb471) had improved half-lives compared to dupilumab across different administration routes (Tables 17, Figure 5A and Figure 5B When administered at 5 and 1 mg / kg IV, construct 38 maintained a half-life superior to dupilumab at 25 mg / kg IV. The increased half-life of the exemplary antibodies may allow for less frequent dosing compared to currently available treatment regimens, thereby reducing the injection burden and increasing compliance for patients with COPD and other diseases.

[0528] Where bioavailability (F) was determined, the antibodies disclosed herein were shown to have bioavailability equivalent to that of dupilumab. The results for exemplary antibodies and dupilumab are summarized in Table 18. Construct 38 (mAb471) showed bioavailability comparable to that of dupilumab.

[0529] Table 18

[0530]

[0531]

[0532] Example 12. Grouping experiment using anti-IL-4Rα antibody (dupirocin) gExperiment)

[0533] Epitope grouping is a technique used to cluster different mAbs based on the specific region of the antigen (in this case IL-4Rα) recognized by the antibodies. In the group study using immobilized dupilumab, no responses were observed for constructs mAb410, mAb411, mAb412, mAb413, mAb414, mAb415, mAb416, mAb417, mAb418, mAb419, mAb420, mAb421, mAb422, mAb422B, mAb423, mAb424, mAb425, mAb426, mAb427, mAb428, mAb429, mAb431, mAb432, mAb433, mAb434, mAb435, mAb436, mAb437, mAb439, mAb443, mAb450, mAb470, mAb471, and mAb476. This indicates that the described mAbs are grouped together with dupilumab and provides evidence that the mAbs likely bind to similar or identical epitopes on IL-4Rα and therefore they are more likely to have the same biological effect.

[0534] Example 13. Half-life estimation

[0535] The half-life of an antibody is a product of degradation or elimination via three pathways: pinocytosis, target-mediated drug disposition (TMDD), and receptor-mediated endocytosis.

[0536] Pinocytosis: Pinocytosis is a nonspecific process in which extracellular fluids and substances enter the cell, creating an internalized vesicle. This internal vesicle then fuses with a lysosome. All antibodies undergo this elimination pathway.

[0537] TMDD: This is a receptor-mediated endocytosis process, meaning that the interaction of an antibody with its receptor leads to the internalization of the antibody-receptor complex and its subsequent degradation via lysosomes, specialized organelles, or intracellular compartments that degrade molecules and other biological materials. TMDD is commonly observed with mAbs with receptor targets, such as dupilumab, which targets IL-4Rα.

[0538] Receptor-mediated endocytosis: Binding of antibodies to Fc-γ receptors, present on many immune cells, can also trigger an elimination process similar to TMDD. However, third-party preclinical studies have shown that this degradation pathway plays only a minor role in antibody elimination, if any at all.

[0539] To estimate the half-life of the disclosed antibodies grouped with dupilumab, the VL and VH domains of dupilumab were combined with an IgG1-YTE constant region ("IL-4Rα tool compound") and the half-life in non-human primates (NHPs) was determined. The IL-4Rα tool compound incorporating the YTE amino acid substitution exhibited a half-life of 18.5 days, a 76% increase compared to 10.5 days for dupilumab. Therefore, given the improved half-life of the IL-4Rα tool compound, it is believed that the antibodies disclosed herein will have significantly improved dosing compared to prior art antibodies (e.g., dupilumab).

[0540] As described above, antibody recycling via increased affinity for FcRn influences degradation via pinocytosis rather than elimination via target-mediated drug disposition (TMDD). Therefore, the half-life extension provided by the YTE mutation in antibodies with membrane-bound targets was used in the following analysis.

[0541] As an example, CDX-0159 is an antibody targeting KIT (c-KIT / CD117) receptor tyrosine kinase with a YTE amino acid substitution for half-life extension in current clinical development. In NHPs, CDX-0159 showed a half-life of 22 days compared to 4.8 days for CDX-0158 (a non-half-life extension antibody directed at the same target). Clinically, CDX-0159 showed a half-life of 32 days, indicating an increase of approximately 1.5 times compared to NHP data. In addition, CDX-0159 has demonstrated a human half-life that is approximately five times longer than CDX-0158 (a non-half-life extension antibody directed at the same target) (the half-life of CDX-0159 is 32 days, compared to 6 days for CDX-0158).

[0542] As another example, VRDN-002 is an antibody targeting the anti-IGF-1 receptor with a recycling-based FC modification (i.e., YTE or LS or similar amino acid substitutions) currently in clinical development for half-life extension. In NHPs, VRDN-002 demonstrated a half-life of 14 days, compared to 6.4 days for teprotumumab (a non-half-life-extending antibody directed at the same target). Clinically, VRDN-002 demonstrated a half-life of approximately 30 to 40 days in an interim analysis, representing an approximately two- to three-fold increase compared to NHP data. Furthermore, VRDN-002 has demonstrated a human half-life that is approximately three to four times longer than teprotumumab (a non-half-life-extending antibody directed at the same target) (VRDN-002 half-life of approximately 30 to 40 days, compared to approximately 10 to 11 days for teprotumumab).

[0543] Based on these studies (and depending on the approach taken), the human half-lives of the antibodies disclosed herein are expected to be approximately 30 to 60 days based on a factor of 1.5 to 3 fold going from NHPs into humans, or approximately 45 to 70 days based on a factor of three to five fold going from a non-half-life extended antibody into a half-life extended antibody directed at the same receptor target.

[0544] To further estimate the half-life of the antibodies disclosed herein, a two-compartment model with first-order absorption and parallel linear and Michaelis-Menten elimination (the latter corresponding to the TMDD effect associated with targeting membrane-bound IL4Ra) was constructed to predict the concentrations of both pilumab and the antibodies disclosed herein over time (drug levels). Parameters included an elimination rate (ke) of 0.0447 days-1, a core volume (Vc) of 2.74 L, an absorption rate (ka) of 0.306 days-1, and a bioavailability of 64.2%.

[0545] It is believed that the efficacy in inflammatory diseases such as COPD is due to the 谷 Therefore, based on the model described above, the C of the disclosed antibodies targeted 谷 Equivalent to 300 mg SC every two weeks for maintenance dupilumab C 谷 , which is approximately 75 mg / L. Given the overlapping epitopes of dupilumab and the antibodies grouped with dupilumab disclosed herein and the similarity of potency in multiple in vitro assays, the necessary exposure for potential clinical activity of the antibodies disclosed herein can be predicted. 消除 Based on half-life modeling to maintain antibody concentrations above about 75 mg / L, a half-life of at least 42 days would be required to maintain dosing of the antibodies disclosed herein every six weeks, and a half-life of at least 59 days would be required to maintain dosing of the antibodies disclosed herein every two months.

[0546] Thus, it is contemplated that the antibodies disclosed herein can be administered every six weeks if they exhibit a half-life of at least 42 days, and every two months if they exhibit a half-life of at least 59 days.

[0547] Incorporated by Reference

[0548] The entire disclosure of each of the patent and scientific literature cited herein is incorporated by reference for all purposes.

[0549] equivalent

[0550] The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The foregoing embodiments are therefore to be considered in all respects as illustrative rather than restrictive of the invention described herein. The scope of the invention is therefore indicated by the appended claims rather than the foregoing description, and all changes that come within the meaning and range of equivalents of the claims are intended to be embraced therein.

[0551] Sequence Listing

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Claims

1. An isolated antibody that binds to interleukin (IL)-4 receptor alpha (IL-4Rα), the antibody comprising: a) a variable heavy (VH) chain sequence having three heavy chain CDR sequences: CDR-H1, CDR-H2, and CDR-H3; and b) a variable light (VL) chain sequence having three light chain CDR sequences: CDR-L1, CDR-L2, and CDR-L3; wherein: a. CDR-H1 comprises a sequence selected from the group consisting of SEQ ID NOs: 1-4, 66-70, and 187-191; b. CDR-H2 comprises a sequence selected from the group consisting of SEQ ID NOs: 5-16 and 71-90; c. CDR-H3 comprises a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 17-25 and 92-99, d. CDR-L1 comprises a sequence selected from the group consisting of SEQ ID NOs: 26-40 and 100-107, e. CDR-L2 comprises a sequence selected from the group consisting of SEQ ID NOs: 41-52 and 108-112; and f. CDR-L3 comprises a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 53-65.

2. The isolated antibody of claim 1, wherein the antibody comprises: a. CDR-H1 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 1-4; b. CDR-H2 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 5-16; c. CDR-H3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 17-25, d. CDR-L1 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 26-40, e. CDR-L2 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 41-52; and f. CDR-L3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 53-65.

3. The isolated antibody of claim 1, wherein the antibody comprises: a. CDR-H1 comprising a sequence selected from the group consisting of SEQ ID NOs: 66-70; b. CDR-H2 comprising a sequence selected from the group consisting of SEQ ID NOs: 71-78; c. CDR-H3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 17-25, d. CDR-L1 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 26-40, e. CDR-L2 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 41-52; and f. CDR-L3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 53-65.

4. The isolated antibody of claim 1, wherein the antibody comprises: a. CDR-H1 comprising a sequence selected from the group consisting of SEQ ID NOs: 187-191; b. CDR-H2 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 79-90; c. CDR-H3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 92-99, d. CDR-L1 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 100-107, e. CDR-L2 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 108-112; and f. CDR-L3 comprising a sequence selected from the group consisting of sequences shown in SEQ ID NOs: 53-65.

5. The isolated antibody of any one of the above claims, wherein the antibody does not comprise: a. CDR-H1 as shown in SEQ ID NO: 1; CDR-H2 as shown in SEQ ID NO: 5; CDR-H3 as shown in SEQ ID NO: 17; CDR-L1 as shown in SEQ ID NO: 26; CDR-L2 as shown in SEQ ID NO: 41; and CDR-L3 as shown in SEQ ID NO: 53; or b. CDR-H1 shown in SEQ ID NO: 67; CDR-H2 shown in SEQ ID NO: 71; CDR-H3 shown in SEQ ID NO: 17; CDR-L1 shown in SEQ ID NO: 26; CDR-L2 shown in SEQ ID NO: 41; and CDR-L3 shown in SEQ ID NO: 53; or c. CDR-H1 shown in SEQ ID NO: 187; CDR-H2 shown in SEQ ID NO: 79; CDR-H3 shown in SEQ ID NO: 91; CDR-L1 shown in SEQ ID NO: 100; CDR-L2 shown in SEQ ID NO: 108; and CDR-L3 shown in SEQ ID NO:

53.

6. The isolated antibody of claim 5, wherein the antibody does not comprise any combination of: a. CDR-H1 shown in any one of SEQ ID NO: 1, 67 or 187; b. CDR-H2 as shown in any one of SEQ ID NO: 5, 71 or 79; c. CDR-H3 shown in either SEQ ID NO: 17 or 91; d. CDR-L1 shown in any one of SEQ ID NO: 26 or 100; e. CDR-L2 as shown in either SEQ ID NO: 41 or 108; and f. CDR-L3 shown in SEQ ID NO:

53.

7. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

54.

8. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 62, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

55.

9. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

55.

10. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

55.

11. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

56.

12. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

57.

13. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 46 or 109; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

58.

14. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

59.

15. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 68, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

60.

16. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 29 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

60.

17. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

57.

18. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

19. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

20. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

21. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 21 or 96; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

22. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

62.

23. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

24. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 23, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

60.

25. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22 or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

56.

26. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 23 or 97; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

27. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 72, or 81; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20, 18, or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

28. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 7, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

29. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 36 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

30. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 37 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

31. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 12, 74, or 86; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

32. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 189; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 13, 71, or 87; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

33. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 10, 71, or 84; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

57.

34. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

35. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 66, or 187; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 38 or 105; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 48 or 111; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

63.

36. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 41 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

64.

37. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 39 or 106; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 49 or 112; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

63.

38. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

60.

39. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 8, 73, or 82; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 31 or 103; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

58.

40. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 33 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 50 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

41. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

42. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 67, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 67, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 40 or 107; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 52 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

43. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 20 or 95; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 47 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

44. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 11, 73, or 85; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 22, 18, or 93; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 30 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

60.

45. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 14, 75, or 88; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 27 or 101; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

65.

46. ​​The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

62.

47. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 3, 69, or 190; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 9, 77, or 83; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

55.

48. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 15, 76, or 89; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 24 or 98; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 42 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

49. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 6, 71, or 80; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 18 or 92; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 32 or 102; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 51 or 110; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

61.

50. The isolated antibody of any one of claims 1-6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 2, 67, or 188; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 19 or 94; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 28 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 45 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

57.

51. The isolated antibody of any one of claims 1 to 6, wherein the antibody comprises: a CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 4, 70, or 191; a CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 16, 78, or 90; a CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 25 or 99; a CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 34 or 100; a CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: 43 or 108; and a CDR-L3 comprising the sequence set forth in SEQ ID NO:

53.

52. The isolated antibody of any one of the above claims, wherein the antibody comprises a heavy chain variable domain (VH) sequence selected from the group consisting of sequences shown in SEQ ID NOs: 113-145.

53. The isolated antibody of any one of the above claims, wherein the antibody comprises a light chain variable domain (VL) sequence selected from the group consisting of sequences shown in SEQ ID NOs: 146-186.

54. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence selected from the group consisting of sequences shown in SEQ ID NOs: 113-145, and a VL sequence selected from the group consisting of sequences shown in SEQ ID NOs: 146-186.

55. An isolated antibody that binds to IL-4Rα, wherein the antibody comprises a VH sequence selected from the group consisting of sequences shown in SEQ ID NOs: 113-145, and a VL sequence selected from the group consisting of sequences shown in SEQ ID NOs: 146-186.

56. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

147.

57. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

148.

58. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 115 and a VL sequence as shown in SEQ ID NO:

149.

59. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

150.

60. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 116 and a VL sequence as shown in SEQ ID NO:

151.

61. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 117 and a VL sequence as shown in SEQ ID NO:

152.

62. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 118 and a VL sequence as shown in SEQ ID NO:

153.

63. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 117 and a VL sequence as shown in SEQ ID NO:

154.

64. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 119 and a VL sequence as shown in SEQ ID NO:

155.

65. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 120 and a VL sequence as shown in SEQ ID NO:

156.

66. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 120 and a VL sequence as shown in SEQ ID NO:

157.

67. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

158.

68. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 121 and a VL sequence as shown in SEQ ID NO:

158.

69. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 122 and a VL sequence as shown in SEQ ID NO:

158.

70. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

159.

71. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 123 and a VL sequence as shown in SEQ ID NO:

159.

72. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 124 and a VL sequence as shown in SEQ ID NO:

160.

73. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 125 and a VL sequence as shown in SEQ ID NO:

161.

74. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 126 and a VL sequence as shown in SEQ ID NO:

162.

75. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 127 and a VL sequence as shown in SEQ ID NO:

163.

76. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 128 and a VL sequence as shown in SEQ ID NO:

164.

77. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 124 and a VL sequence as shown in SEQ ID NO:

158.

78. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 129 and a VL sequence as shown in SEQ ID NO:

165.

79. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 130 and a VL sequence as shown in SEQ ID NO:

166.

80. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 131 and a VL sequence as shown in SEQ ID NO:

167.

81. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 132 and a VL sequence as shown in SEQ ID NO:

159.

82. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 133 and a VL sequence as shown in SEQ ID NO:

159.

83. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 121 and a VL sequence as shown in SEQ ID NO:

168.

84. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 134 and a VL sequence as shown in SEQ ID NO:

169.

85. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 113 and a VL sequence as shown in SEQ ID NO:

170.

86. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 134 and a VL sequence as shown in SEQ ID NO:

171.

87. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 135 and a VL sequence as shown in SEQ ID NO:

172.

88. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 136 and a VL sequence as shown in SEQ ID NO:

173.

89. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 131 and a VL sequence as shown in SEQ ID NO:

174.

90. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 137 and a VL sequence as shown in SEQ ID NO:

175.

91. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

176.

92. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 138 and a VL sequence as shown in SEQ ID NO:

177.

93. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 134 and a VL sequence as shown in SEQ ID NO:

178.

94. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 139 and a VL sequence as shown in SEQ ID NO:

179.

95. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 140 and a VL sequence as shown in SEQ ID NO:

180.

96. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 141 and a VL sequence as shown in SEQ ID NO:

181.

97. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 142 and a VL sequence as shown in SEQ ID NO:

182.

98. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 143 and a VL sequence as shown in SEQ ID NO:

183.

99. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 114 and a VL sequence as shown in SEQ ID NO:

184.

100. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 144 and a VL sequence as shown in SEQ ID NO:

185.

101. The isolated antibody of claim 54 or 55, wherein the antibody comprises a VH sequence as shown in SEQ ID NO: 145 and a VL sequence as shown in SEQ ID NO:

186.

102. The isolated antibody of any one of the above claims, wherein the antibody is a humanized, fully human, or chimeric antibody.

103. The isolated antibody of any one of the above claims, wherein the antibody is a fully human antibody.

104. The isolated antibody of any one of the above claims, wherein the antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM.

105. The isolated antibody of any of the above claims, wherein the human Fc region comprises a human heavy chain constant region of class IgG and a subclass selected from IgGl, IgG2, IgG3, and IgG4.

106. The isolated antibody of claim 105, wherein the human Fc region comprises a human IgG1 Fc.

107. The isolated antibody of claim 105, wherein the human Fc region comprises a human IgG4 Fc.

108. The isolated antibody of claim 105, wherein the human Fc region comprises a human IgG2 Fc.

109. The isolated antibody of any one of the above claims, wherein the heavy chain comprises a constant heavy chain sequence selected from the group consisting of sequences shown in SEQ ID NOs: 192-235 and 251-407.

110. An isolated antibody that binds to interleukin (IL)-4 receptor alpha (IL-4Rα), the antibody comprising: a) a variable heavy (VH) chain sequence having three heavy chain CDR sequences: CDR-H1, CDR-H2, and CDR-H3; and b) a variable light (VL) chain sequence having three light chain CDR sequences: CDR-L1, CDR-L2, and CDR-L3; wherein the antibody comprises: CDR-H1 comprising the sequence set forth in any one of SEQ ID NOs: 1, 66, or 187; CDR-H2 comprising the sequence set forth in any one of SEQ ID NOs: 5, 71, or 79; CDR-H3 comprising the sequence set forth in any one of SEQ ID NOs: 17 or 91; CDR-L1 comprising the sequence set forth in any one of SEQ ID NOs: 26 or 100; CDR-L2 comprising the sequence set forth in any one of SEQ ID NOs: NO:41 or 108; and CDR-L3 comprising the sequence shown in SEQ ID NO:53; and wherein the heavy chain comprises a constant heavy chain sequence selected from the group consisting of the sequences shown in SEQ ID NO:192-235 and 251-407.

111. The isolated antibody of claim 110, wherein the antibody comprises the VH sequence shown in SEQ ID NO:

113.

112. The isolated antibody of claim 110 or 111, wherein the antibody comprises the VL sequence shown in SEQ ID NO:

146.

113. The isolated antibody of any one of the above claims, wherein the light chain comprises a constant light chain sequence comprising the sequence shown in SEQ ID NO:

236.

114. The isolated antibody of any one of the above claims, wherein the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions increase one or more of antibody half-life, ADCC activity, ADCP activity, or CDC activity compared to an Fc without the one or more substitutions.

115. The isolated antibody of any one of claims 1-113, wherein the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in a decrease in one or more of ADCC activity, ADCP activity, or CDC activity compared to an antibody comprising a wild-type Fc region.

116. The isolated antibody of claim 114, wherein the one or more amino acid substitutions are selected from the group consisting of: S228P, M252Y, S254T, T256E, T256D, T250Q, H285D, T307A, T307Q, T307R, T307W, L309D, Q411H, Q311V, A378V, E380A, M428L, N434A, N434S; optionally, wherein the one or more amino acid substitutions comprise a plurality of amino acid substitutions selected from the group consisting of: i) M428 L / N434S; ii) M252Y / S254T / T256E; iii) T250Q / M428L; iv) T307A / E380A / N434A; v) T256D / T307Q; vi) T256D / T307W; vii) M2 52Y / T256D; viii) T307Q / Q311V / A378V; ix) T256D / H285D / T307R / Q311V / A378V; x) L309D / Q311H / N434S; and xi) S228P / L235E.

117. The isolated antibody of any one of the above claims, wherein the Fc region binds to the neonatal Fc receptor (FcRn).

118. The isolated antibody of claim 117, wherein the Fc region binds FcRn with a higher affinity at pH 6.0 than an antibody comprising a wild-type Fc region.

119. The isolated antibody of claim 117 or 118, wherein the Fc region is expressed at a pH of <1 x 10 -7 M's K D Binds to FcRn.

120. The isolated antibody of any one of the above claims, wherein the antibody is a monoclonal antibody.

121. The antibody of any of the above claims, wherein the antibody binds to the IL-4Rα sequence shown in SEQ ID NOs: 237-240, 245-247, and 258.

122. The isolated antibody of any one of the above claims, wherein the antibody has a specific activity at a concentration of less than or equal to about 1 x 10 -9 M, 2x 10 -9 M, 3x 10 -9 M, 4x 10 -9 M, 5x 10 -9 M, 6x 10 -9 M, 7x 10 -9 M, 8x 10 -9 M, 9x 10 -9 M's K D Binds to the IL-4Rα sequences shown in SEQ ID NOs: 237-240, 245-247, and 258.

123. The isolated antibody of claim 122, wherein the antibody has a specific activity at a concentration of less than or equal to about 1 x 10 -10 M's K D Binds to the IL-4Rα sequences shown in SEQ ID NOs: 237-240, 245-247, and 258.

124. The isolated antibody of claim 121, wherein the antibody has a specific activity at a concentration of less than or equal to about 1 x 10 -9 M's K D Binds to human IL-4Rα.

125. The isolated antibody of any one of the above claims, wherein the antibody exhibits a melting temperature greater than 68°C as measured by differential scanning fluorimetry (DSF).

126. The isolated antibody of claim 125, wherein the antibody exhibits a melting temperature greater than 75°C as measured by differential scanning fluorimetry (DSF).

127. The isolated antibody of any one of the above claims, wherein the antibody exhibits an aggregation temperature equal to or greater than 71.2°C as measured by differential scanning fluorimetry (DSF).

128. The isolated antibody of any one of the above claims, wherein the antibody has a retention time of 15.2 minutes or less as measured by hydrophobic interaction chromatography.

129. The isolated antibody of any one of claims 1-110 and 112-128, wherein the antibody does not have the heavy chain sequence shown in SEQ ID NO.

113.

130. The isolated antibody of any one of claims 1-111 and 113-129, wherein the antibody does not have the heavy chain sequence shown in SEQ ID NO.

146.

131. The isolated antibody of any of the above claims for use in treating an inflammatory disorder or disease.

132. The isolated antibody of claim 131 , for use in treating atopic dermatitis (AD).

133. The isolated antibody of claim 132, wherein the treatment reduces disease severity in the patient, and wherein disease severity is assessed by the Atopic Dermatitis Disease Severity Outcome Index.

134. The isolated antibody of claim 131 , for use in treating asthma.

135. The isolated antibody of claim 132, for use in treating chronic sinusitis with nasal polyps.

136. The isolated antibody of claim 131, for use in treating chronic rhinosinusitis without nasal polyps (CRSsNP).

137. The isolated antibody of claim 131, for use in treating eosinophilic esophagitis (EoE).

138. The isolated antibody of claim 131 , for use in treating an eosinophilic gastrointestinal disorder or disease (ENID) selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC), and eosinophilic gastroenteritis (EGE).

139. The isolated antibody of claim 131, for use in treating Chargar-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA).

140. The isolated antibody of claim 131, for use in treating prurigo nodularis (PN).

141. The isolated antibody of claim 131, for use in treating chronic spontaneous urticaria (CSU).

142. The isolated antibody of claim 131, for use in treating chronic pruritus of unknown origin (CPUO).

143. The isolated antibody of claim 131, for use in treating bullous pemphigoid (BP).

144. The isolated antibody of claim 131 , for use in treating cold-induced urticaria (ColdU).

145. The isolated antibody of claim 131, for use in treating allergic fungal sinusitis (AFRS).

146. The isolated antibody of claim 131, for use in treating allergic bronchopulmonary aspergillosis.

147. The isolated antibody of claim 131, for use in treating chronic obstructive pulmonary disease (COPD).

148. An isolated polynucleotide or group of polynucleotides, which encodes the antibody, its VH, its VL, its light chain, its heavy chain or its antigen binding portion according to any of the above claims, and optionally, wherein the polynucleotide or group of polynucleotides comprises cDNA.

149. A vector or vector set comprising the polynucleotide or polynucleotide set of claim 148.

150. A host cell comprising the polynucleotide or set of polynucleotides according to claim 148 or the vector or set of vectors according to claim 149.

151. A method of producing an antibody, the method comprising expressing the antibody using the host cell of claim 150 and isolating the expressed antibody.

152. A pharmaceutical composition comprising the antibody of any one of claims 1-147 and a pharmaceutically acceptable excipient.

153. A kit comprising the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152 and instructions for use.

154. A method for treating an inflammatory condition or disease in a mammalian subject in need thereof, comprising administering to the mammalian subject a therapeutically effective amount of the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152.

155. The method of claim 154, wherein the inflammatory disorder or disease is atopic dermatitis.

156. The method of claim 154, wherein the inflammatory disorder or disease is asthma.

157. The method of claim 154, wherein the inflammatory condition or disease is chronic sinusitis with nasal polyps.

158. The method of claim 154, wherein the inflammatory condition or disease is chronic rhinosinusitis without nasal polyps (CRSsNP).

159. The method of claim 154, wherein the inflammatory disorder or disease is eosinophilic esophagitis (EoE).

160. The method of claim 154, wherein the inflammatory disorder or disease is an eosinophilic gastrointestinal disorder or disease (ENID) selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC), and eosinophilic gastroenteritis (EGE).

161. The method of claim 154, wherein the inflammatory disorder or disease is Chargar-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA).

162. The method of claim 154, wherein the inflammatory disorder or disease is prurigo nodularis (PN).

163. The method of claim 154, wherein the inflammatory disorder or disease is chronic spontaneous urticaria (CSU).

164. The method of claim 154, wherein the inflammatory disorder or disease is chronic pruritus of unexplained origin (CPUO).

165. The method of claim 154, wherein the inflammatory disorder or disease is bullous pemphigoid (BP).

166. The method of claim 154, wherein the inflammatory disorder or disease is cold-induced urticaria (ColdU).

167. The method of claim 154, wherein the inflammatory condition or disease is allergic fungal sinusitis (AFRS).

168. The method of claim 154, wherein the inflammatory disorder or disease is allergic bronchopulmonary aspergillosis (ABPA).

169. The method of claim 154, wherein the inflammatory disorder or disease is chronic obstructive pulmonary disease (COPD).

170. A method for treating a pathology associated with elevated levels of IL-4 and / or IL-13 in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152.

171. A method of reducing the biological activity of IL-4, IL-13 and / or IL-4Rα in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152.

172. A method of inhibiting a TH2-type allergic response in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152.

173. A method of preventing an inflammatory condition or disease in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of the antibody of any one of claims 1-147 or the pharmaceutical composition of claim 152.

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