Antibodies that bind to interleukin 13 and methods of use

Abstract

This specification describes novel and improved antibodies that bind to interleukin 13 (IL-13) and methods of using the same. In certain embodiments, this specification describes methods of inhibiting the biological activity of IL-13. In certain embodiments, this specification describes pharmaceutical compositions comprising anti-IL-13 antibodies. In certain embodiments, the antibodies and methods described herein are used for the treatment of inflammatory diseases or disorders associated with elevated levels of IL-13 and / or IgE.

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit and priority of U.S. Provisional Application No. 63 / 353,367, filed on June 17, 2022, U.S. Provisional Application No. 63 / 462,822, filed on April 28, 2023, and U.S. Provisional Application No. 63 / 469,167, filed on May 26, 2023. The entire disclosure of the provisional applications is hereby incorporated by reference in its entirety for all purposes.

Background Art

[0002] Interleukin (IL)-13 is a T helper cell subclass 2 (Th2) cytokine that belongs to the type I cytokine family and exhibits pleiotropic effects across multiple cellular pathways. IL-13 is involved in the differentiation of naive T cells into Th2 cells. IL-13 promotes B cell proliferation and induces isotype class switching of immunoglobulins to IgG4 and IgE when co-stimulated with CD40 / CD40L. It also upregulates FcεRI and thus aids in IgE priming of mast cells. In monocytes / macrophages, IL-13 upregulates the expression of CD23 and MHC class I and class II antigens, downregulates the expression of CD14, inhibits antibody-dependent cell cytotoxicity, and promotes eosinophil survival, activation, and recruitment. IL-13 also exhibits important functions in non-hematopoietic cells, such as smooth muscle cells, epithelial cells, endothelial cells, and fibroblasts. IL-13 enhances smooth muscle proliferation and cholinergic-induced contraction. In epithelial cells, IL-13 is a potent inducer of chemokine production, alters mucociliary differentiation, decreases the ciliary beat frequency of ciliated epithelial cells, and results in goblet cell metaplasia. In endothelial cells, IL-13 is a potent inducer of vascular cell adhesion molecule 1 (VCAM-1), which is important for eosinophil recruitment. In epithelial keratinocytes, IL-13 decreases the expression of barrier integrity molecules, such as filaggrin and loricrin, while stimulating the secretion of CCL26 and CCL2, which are involved in the recruitment of some inflammatory cells of the myeloid cell lineage. In human skin fibroblasts, IL-13 induces type I collagen synthesis in human skin fibroblasts.

[0003] Inhibition of IL-13 can be used to treat or prevent diseases and conditions associated with elevated IgE levels, including, but not limited to, inflammatory diseases and conditions such as asthma, allergic rhinitis, urticaria, and allergic or atopic dermatitis. Therefore, the development of potent and specific inhibitors of IL-13, such as inhibitors that maintain activity for a longer period when administered to a subject, is needed for the prevention and / or treatment of IL-13- and IgE-mediated diseases or conditions. SUMMARY OF THE INVENTION

[0004] In certain embodiments, described herein is an isolated antibody that binds to IL-13 and includes i) a variable heavy (VH) chain sequence comprising three heavy chain CDR sequences, namely CDR-H1, CDR-H2, and CDR-H3, and ii) a variable light (VL) chain sequence comprising three light chain CDR sequences, namely CDR-L1, CDR-L2, and CDR-L3, wherein a) CDR-H1 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 58-99 and 121; b) CDR-H2 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 100-111; c) CDR-H3 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 112-120 and 130-140; d) CDR-L1 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 141-144 and 149-152; e) CDR-L2 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 153-158 and the amino acid sequence LAS; and f) CDR-L3 comprises a sequence selected from the sequences set forth in SEQ ID NOs: 165-172.

[0005] In certain embodiments, the isolated antibody includes a) CDR-H1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 58-66; b) CDR-H2 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 100-103; c) CDR-H3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 112-120; d) CDR-L1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 141-144; e) CDR-L2 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 153-158; and f) CDR-L3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 165-172.

[0006] In certain embodiments, the isolated antibody comprises: a) CDR-H1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 67-83; b) CDR-H2 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 104-107; c) CDR-H3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 112-120; d) CDR-L1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 141-144; e) CDR-L2 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 153-158; and f) CDR-L3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 165-172.

[0007] In certain embodiments, the isolated antibody comprises: a) CDR-H1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 84-99 and 121; b) CDR-H2 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 108-111; c) CDR-H3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 130-140; d) CDR-L1 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 149-152; e) CDR-L2 comprising the amino acid sequence LAS; and f) CDR-L3 comprising a sequence selected from the sequences set forth in SEQ ID NOs: 165-172.

[0008] In certain embodiments, the isolated antibody does not comprise: a) CDR-H1 set forth in SEQ ID NO: 58, CDR-H2 set forth in SEQ ID NO: 100, CDR-H3 set forth in SEQ ID NO: 112, CDR-L1 set forth in SEQ ID NO: 141, CDR-L2 set forth in SEQ ID NO: 153, and CDR-L3 set forth in SEQ ID NO: 165; or b) CDR-H1 set forth in SEQ ID NO: 67, CDR-H2 set forth in SEQ ID NO: 104, CDR-H3 set forth in SEQ ID NO: 112, CDR-L1 set forth in SEQ ID NO: 141, CDR-L2 set forth in SEQ ID NO: 153, and CDR-L3 set forth in SEQ ID NO: 165; or c) CDR-H1 set forth in SEQ ID NO: 84, CDR-H2 set forth in SEQ ID NO: 108, CDR-H3 set forth in SEQ ID NO: 130, CDR-L1 set forth in SEQ ID NO: 149, CDR-L2 represented by the amino acid sequence LAS, and CDR-L3 set forth in SEQ ID NO: 165.

[0009] In certain embodiments, the antibody does not include any combination of a) CDR-H1 as set forth in any of SEQ ID NO: 58, 67, or 84, b) CDR-H2 as set forth in any of SEQ ID NO: 100, 104, or 108, c) CDR-H3 as set forth in either SEQ ID NO: 112 or 130, d) CDR-L1 as set forth in either SEQ ID NO: 141 or 149, e) CDR-L2 as set forth in either SEQ ID NO: 153 or 154, and f) CDR-L3 as set forth in SEQ ID NO: 165.

[0010] In certain embodiments, the isolated antibody comprises a CDR-H1 comprising the sequence set forth in any of SEQ ID NO: 58, 67, or 68, a CDR-H2 comprising the sequence set forth in any of SEQ ID NO: 100 or 104, a CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, a CDR-L1 comprising the sequence set forth in either SEQ ID NO: 141 or 149, a CDR-L2 comprising the sequence set forth in either SEQ ID NO: 153 or the amino acid sequence LAS, and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0011] In certain embodiments, the isolated antibody comprises a CDR-H1 comprising the sequence set forth in SEQ ID NO: 58, a CDR-H2 comprising the sequence set forth in SEQ ID NO: 100, a CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, a CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, a CDR-L2 comprising the sequence set forth in SEQ ID NO: 153, and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0012] In certain embodiments, the isolated antibody comprises a CDR-H1 comprising the sequence set forth in SEQ ID NO: 67, a CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, a CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, a CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, a CDR-L2 comprising the sequence set forth in SEQ ID NO: 153, and a CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0013] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 68, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, CDR-L2 comprising the sequence set forth in SEQ ID NO: 153, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0014] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 67, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0015] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 68, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0016] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in any of SEQ ID NOs: 58, 67, 68, 84, or 85, CDR-H2 comprising the sequence set forth in any of SEQ ID NOs: 100, 104, or 108, CDR-H3 comprising the sequence set forth in either of SEQ ID NOs: 112 or 130, CDR-L1 comprising the sequence set forth in either of SEQ ID NOs: 141 or 149, CDR-L2 comprising the sequence set forth in either of SEQ ID NO: 153 or the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0017] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 68, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, CDR-L2 comprising the sequence set forth in SEQ ID NO: 153, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0018] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 84, CDR-H2 comprising the sequence set forth in SEQ ID NO: 108, CDR-H3 comprising the sequence set forth in SEQ ID NO: 130, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0019] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 85, CDR-H2 comprising the sequence set forth in SEQ ID NO: 108, CDR-H3 comprising the sequence set forth in SEQ ID NO: 130, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0020] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in any of SEQ ID NO: 58, 67, 68, 84, or 85, CDR-H2 comprising the sequence set forth in any of SEQ ID NO: 100, 104, or 108, CDR-H3 comprising the sequence set forth in either SEQ ID NO: 112 or 130, CDR-L1 comprising the sequence set forth in either SEQ ID NO: 141 or 149, CDR-L2 comprising the sequence set forth in either SEQ ID NO: 157 or the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0021] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 58, CDR-H2 comprising the sequence set forth in SEQ ID NO: 100, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, CDR-L2 comprising the sequence set forth in SEQ ID NO: 157, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0022] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 68, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, CDR-L2 comprising the sequence set forth in SEQ ID NO: 157, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0023] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in any of SEQ ID NO: 58, 67, 68, 84, or 85, CDR-H2 comprising the sequence set forth in any of SEQ ID NO: 100, 104, or 108, CDR-H3 comprising the sequence set forth in either SEQ ID NO: 112 or 130, CDR-L1 comprising the sequence set forth in either SEQ ID NO: 141 or 149, CDR-L2 comprising the sequence set forth in either SEQ ID NO: 157 or the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0024] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 68, CDR-H2 comprising the sequence set forth in SEQ ID NO: 104, CDR-H3 comprising the sequence set forth in SEQ ID NO: 112, CDR-L1 comprising the sequence set forth in SEQ ID NO: 141, CDR-L2 comprising the sequence set forth in SEQ ID NO: 157, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0025] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 84, CDR-H2 comprising the sequence set forth in SEQ ID NO: 108, CDR-H3 comprising the sequence set forth in SEQ ID NO: 130, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0026] In certain embodiments, the isolated antibody comprises CDR-H1 comprising the sequence set forth in SEQ ID NO: 85, CDR-H2 comprising the sequence set forth in SEQ ID NO: 108, CDR-H3 comprising the sequence set forth in SEQ ID NO: 130, CDR-L1 comprising the sequence set forth in SEQ ID NO: 149, CDR-L2 comprising the amino acid sequence LAS, and CDR-L3 comprising the sequence set forth in SEQ ID NO: 165.

[0027] In certain embodiments, the isolated antibody comprises a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470.

[0028] In certain embodiments, the isolated antibody comprises a VL sequence selected from the sequences set forth in SEQ ID NOs: 33-57 and 471.

[0029] In certain embodiments, the isolated antibody comprises a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence selected from the sequences set forth in SEQ ID NOs: 33-57 and 471.

[0030] In certain embodiments, the isolated antibody comprises a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence comprising the sequence set forth in SEQ ID NO: 49.

[0031] In certain embodiments, the isolated antibody comprises a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence comprising the sequence set forth in SEQ ID NO: 51.

[0032] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 1 and the VL sequence set forth in SEQ ID NO: 33.

[0033] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 2 and the VL sequence set forth in SEQ ID NO: 33.

[0034] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 35.

[0035] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 35.

[0036] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 35.

[0037] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 35.

[0038] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 35.

[0039] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 36.

[0040] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 36.

[0041] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 36.

[0042] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 36.

[0043] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 36.

[0044] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39.

[0045] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 39.

[0046] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 39.

[0047] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 39.

[0048] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39.

[0049] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 40.

[0050] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 40.

[0051] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 40.

[0052] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 40.

[0053] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 40.

[0054] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 42.

[0055] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 9 and the VL sequence set forth in SEQ ID NO: 43.

[0056] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39.

[0057] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 44.

[0058] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 45.

[0059] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 46.

[0060] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 47.

[0061] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 48.

[0062] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 49.

[0063] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 50.

[0064] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51.

[0065] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51.

[0066] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 52.

[0067] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 53.

[0068] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 54.

[0069] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 55.

[0070] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 56.

[0071] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 57.

[0072] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 10 and the VL sequence set forth in SEQ ID NO: 39.

[0073] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 11 and the VL sequence set forth in SEQ ID NO: 39.

[0074] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 12 and the VL sequence set forth in SEQ ID NO: 39.

[0075] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 13 and the VL sequence set forth in SEQ ID NO: 39.

[0076] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 14 and the VL sequence set forth in SEQ ID NO: 39.

[0077] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39.

[0078] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 16 and the VL sequence set forth in SEQ ID NO: 39.

[0079] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 17 and the VL sequence set forth in SEQ ID NO: 39.

[0080] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 18 and the VL sequence set forth in SEQ ID NO: 39.

[0081] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 19 and the VL sequence set forth in SEQ ID NO: 39.

[0082] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 20 and the VL sequence set forth in SEQ ID NO: 39.

[0083] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 21 and the VL sequence set forth in SEQ ID NO: 39.

[0084] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 22 and the VL sequence set forth in SEQ ID NO: 39.

[0085] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 23 and the VL sequence set forth in SEQ ID NO: 39.

[0086] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 24 and the VL sequence set forth in SEQ ID NO: 39.

[0087] The isolated antibody according to claim 21, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 25 and the VL sequence set forth in SEQ ID NO: 39.

[0088] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 26 and the VL sequence set forth in SEQ ID NO: 39.

[0089] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 27 and the VL sequence set forth in SEQ ID NO: 39.

[0090] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39.

[0091] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39.

[0092] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 29 and the VL sequence set forth in SEQ ID NO: 39.

[0093] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 30 and the VL sequence set forth in SEQ ID NO: 39.

[0094] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 31 and the VL sequence set forth in SEQ ID NO: 39.

[0095] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 32 and the VL sequence set forth in SEQ ID NO: 39.

[0096] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39.

[0097] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51.

[0098] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471.

[0099] In certain embodiments, the isolated antibody is a humanized, human, or chimeric antibody. In certain embodiments, the isolated antibody is a humanized antibody. In certain embodiments, the isolated antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM. In certain embodiments, the human Fc region comprises a human heavy chain constant region of a subclass selected from class IgG and IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the human Fc region comprises the Fc of human IgG1. In certain embodiments, the human Fc region comprises the Fc of human IgG4. In certain embodiments, the human Fc region comprises the Fc of human IgG2.

[0100] In certain embodiments of the antibodies described herein, the heavy chain comprises a constant heavy chain sequence selected from the sequences set forth in SEQ ID NOs: 425-468 and 484-539.

[0101] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 1 and the VL sequence set forth in SEQ ID NO: 33, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0102] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 2 and the VL sequence set forth in SEQ ID NO: 33, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0103] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0104] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0105] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0106] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0107] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0108] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0109] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0110] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0111] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0112] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 439, 440, 446, 457, and 460.

[0113] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0114] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0115] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0116] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0117] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0118] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0119] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0120] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0121] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0122] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0123] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 42, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0124] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 9 and the VL sequence set forth in SEQ ID NO: 43, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0125] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0126] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 44, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0127] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 45, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0128] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 46, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0129] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 47, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0130] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 48, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0131] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 49, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0132] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 50, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0133] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0134] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0135] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 52, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0136] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 53, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0137] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 54, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0138] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 55, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0139] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:7 and the VL sequence set forth in SEQ ID NO:56, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0140] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:7 and the VL sequence set forth in SEQ ID NO:57, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0141] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:10 and the VL sequence set forth in SEQ ID NO:39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0142] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:11 and the VL sequence set forth in SEQ ID NO:39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0143] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:12 and the VL sequence set forth in SEQ ID NO:39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0144] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO:13 and the VL sequence set forth in SEQ ID NO:39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0145] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 14 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0146] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0147] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 16 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0148] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 17 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0149] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 18 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0150] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 19 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0151] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 20 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0152] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 21 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0153] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 22 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0154] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 23 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0155] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 24 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0156] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 25 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0157] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 26 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0158] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 27 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0159] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0160] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NOs: 436-468 and 484-539.

[0161] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 29 and a VL sequence selected from the sequences set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NOs: 436-468 and 484-539.

[0162] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 30 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0163] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 31 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0164] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 32 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOS: 436-468 and 484-539.

[0165] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOS: 436-468 and 484-539.

[0166] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOS: 436-468 and 484-539. In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOS: 436-468 and 484-539.

[0167] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439.

[0168] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446.

[0169] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457.

[0170] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 460.

[0171] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 439.

[0172] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 446.

[0173] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 457.

[0174] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 460.

[0175] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 439.

[0176] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 446.

[0177] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 457.

[0178] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 460.

[0179] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 439.

[0180] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 446.

[0181] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 457.

[0182] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 460.

[0183] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 439.

[0184] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446.

[0185] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457.

[0186] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460.

[0187] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439.

[0188] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457.

[0189] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460.

[0190] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446.

[0191] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439.

[0192] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457.

[0193] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460.

[0194] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446.

[0195] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439.

[0196] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457.

[0197] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460.

[0198] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446.

[0199] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439.

[0200] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457.

[0201] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460.

[0202] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446.

[0203] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439.

[0204] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457.

[0205] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460.

[0206] In certain embodiments, the isolated antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446. In certain embodiments, the light chain of the isolated antibody comprises the constant light chain sequence represented by SEQ ID NO: 469.

[0207] In certain embodiments, the Fc region of the isolated antibody comprises one or more amino acid substitutions, which result in a change (e.g., an increase or decrease) in the half-life, ADCC activity, ADCP activity, or CDC activity of the antibody as compared to an otherwise equivalent antibody that does not contain the one or more substitutions. In certain embodiments, the change is an extension of the half-life of the antibody, an increase or decrease in ADCC activity, an increase in ADCP activity, or an increase in CDC activity as compared to an otherwise equivalent antibody that does not contain the one or more substitutions. In certain embodiments, the one or more amino acid substitutions result in an extension of the half-life of the antibody as compared to an antibody comprising a wild-type Fc region. In certain embodiments, the isolated antibody comprising an Fc region having the one or more amino acid substitutions has a half-life of about 80 to 110 days in humans.

[0208] In certain embodiments, the change is an increase or decrease in the half-life of the antibody, an increase or decrease in ADCC activity, an increase or decrease in ADCP activity, or an increase or decrease in CDC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in the half-life of the antibody as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is a decrease in the half-life of the antibody as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in ADCC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is a decrease in ADCC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in ADCP activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is a decrease in ADCP activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in CDC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is a decrease in CDC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in the half-life of the antibody, an increase in ADCC activity, an increase in ADCP activity, and an increase in CDC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions. In certain embodiments, the change is an increase in the half-life of the antibody, a decrease in ADCC activity, an increase in ADCP activity, and an increase in CDC activity as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions.

[0209] In certain embodiments, the change is an increase in the half-life of the antibody as compared to an equivalent antibody except that the equivalent antibody contains an Fc that does not contain the one or more substitutions.

[0210] In certain embodiments, the change is an increase in ADCC activity as compared to an equivalent antibody except that it comprises an Fc that does not contain the one or more substitutions.

[0211] In certain embodiments, the change is a decrease in ADCC activity as compared to an equivalent antibody except that it comprises an Fc that does not contain the one or more substitutions.

[0212] In certain embodiments, the Fc region binds to the neonatal Fc receptor (FcRn). In certain embodiments, the Fc region binds to FcRn with a higher affinity as compared to an antibody comprising a wild-type Fc region at pH 6.0. In certain embodiments, the Fc region binds to FcRn at pH 6.0 with a K D <1x10 -7 M.

[0213] In certain embodiments, the isolated antibody is a monoclonal antibody.

[0214] In certain embodiments, the antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475.

[0215] In certain embodiments, the isolated antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475 with a K as measured by surface plasmon resonance (SPR) of D about 1, 2, 3, 4, 5, 6, 7, 8, 9x10 -9 M or less. In certain embodiments, the isolated antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475 with a K as measured by SPR of D about 1x10 -10 M or less. In certain embodiments, the antibody binds to human IL-13 with a K as measured by SPR of D about 1x10 -9 M or less.

[0216] In certain embodiments, the isolated antibody exhibits a melting temperature of greater than 68 °C as measured by differential scanning fluorimetry (DSF). In certain embodiments, the antibody exhibits a melting temperature of greater than 75 °C as measured by DSF. In certain embodiments, the antibody exhibits an aggregation temperature of 71.2 °C or higher as measured by DSF.

[0217] In certain embodiments, the isolated antibody has a retention time of 15.2 minutes or less as measured by hydrophobic interaction chromatography.

[0218] In certain embodiments, the isolated antibody does not have the heavy chain variable region sequence set forth in SEQ ID NO: 470.

[0219] In certain embodiments, the isolated antibody is used for the treatment of an inflammatory disorder or disease. In certain embodiments, the isolated antibody is used for the treatment of atopic dermatitis. In certain embodiments, the treatment reduces the severity of the disease in a subject, where the severity of the disease is evaluated by the Atopic Dermatitis Disease Severity Outcome Measure. In certain embodiments, the isolated antibody is used for the treatment of asthma. In certain embodiments, the isolated antibody is used for the treatment of idiopathic pulmonary fibrosis. In certain embodiments, the isolated antibody is used for the treatment of alopecia areata. In certain embodiments, the isolated antibody is used for the treatment of chronic rhinosinusitis with nasal polyps. In certain embodiments, the isolated antibody is used for the treatment of chronic rhinosinusitis without nasal polyps (CRSsNP). In certain embodiments, the isolated antibody is used for the treatment of eosinophilic esophagitis (EoE). In certain embodiments, the isolated antibody is used for the treatment of 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 isolated antibody is used for the treatment of Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA). In certain embodiments, the isolated antibody is used for the treatment of prurigo nodularis (PN). In certain embodiments, the isolated antibody is used for the treatment of chronic spontaneous urticaria (CSU). In certain embodiments, the isolated antibody is used for the treatment of chronic pruritus of unknown origin (CPUO). In certain embodiments, the isolated antibody is used for the treatment of bullous pemphigoid (BP). In certain embodiments, the isolated antibody is used for the treatment of cold urticaria (ColdU). In certain embodiments, the isolated antibody is used for the treatment of allergic fungal rhinosinusitis (AFRS). In certain embodiments, the isolated antibody is used for the treatment of allergic bronchopulmonary aspergillosis (ABPA). In certain embodiments, the isolated antibody is used for the treatment of chronic obstructive pulmonary disease (COPD).In certain embodiments, the isolated antibody is used for the treatment of inflammatory bowel diseases such as Crohn's disease or ulcerative colitis. In certain embodiments, the isolated antibody is used for the treatment of psoriasis. In certain embodiments, the isolated antibody is used for the treatment of lupus. In certain embodiments, the isolated antibody is used for the treatment of rheumatoid arthritis.

[0220] In certain aspects, described herein is an isolated polynucleotide or set of polynucleotides encoding the antibody described herein, its VH, its VL, its light chain, its heavy chain, or its antigen-binding portion, optionally wherein the polynucleotide or set of polynucleotides comprises cDNA. In certain aspects, described herein is a vector or set of vectors comprising the polynucleotide or set of polynucleotides. In certain aspects, described herein is a host cell comprising the polynucleotide or set of polynucleotides or the vector or set of vectors.

[0221] In certain aspects, described herein is a method for producing an antibody, the method comprising expressing the antibody using a host cell described herein and isolating the expressed antibody.

[0222] In certain aspects, described herein is a pharmaceutical composition comprising the antibody described herein and a pharmaceutically acceptable excipient.

[0223] In certain aspects, described herein is a kit comprising the antibody described herein or the pharmaceutical composition described herein and instructions for use.

[0224] In certain embodiments, described herein is a method of treating a mammalian subject in need of treatment for an inflammatory disorder or disease, 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 disorder or disease is atopic dermatitis. In certain embodiments, the inflammatory disorder or disease is asthma. In certain embodiments, the inflammatory disorder or disease is idiopathic pulmonary fibrosis. In certain embodiments, the inflammatory disorder or disease is alopecia areata. In certain embodiments, the inflammatory disorder or disease is chronic rhinosinusitis with nasal polyps. In certain embodiments, the inflammatory disorder or disease is chronic rhinosinusitis without nasal polyps (CRSsNP). In certain embodiments, the inflammatory disorder or disease is eosinophilic esophagitis (EoE). In certain embodiments, 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). In certain embodiments, the inflammatory disorder or disease is Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA). In certain embodiments, the inflammatory disorder or disease is prurigo nodularis (PN). In certain embodiments, the inflammatory disorder or disease is chronic spontaneous urticaria (CSU). In certain embodiments, the inflammatory disorder or disease is chronic pruritus of unknown origin (CPUO). In certain embodiments, the inflammatory disorder or disease is bullous pemphigoid (BP). In certain embodiments, the inflammatory disorder or disease is cold urticaria (ColdU). In certain embodiments, the inflammatory disorder or disease is allergic fungal rhinosinusitis (AFRS). In certain embodiments, the inflammatory disorder or disease is allergic bronchopulmonary aspergillosis (ABPA). In certain embodiments, the inflammatory disorder or disease is chronic obstructive pulmonary disease (COPD).In certain embodiments, the inflammatory disorder or disease is an inflammatory bowel disease, such as Crohn's disease or ulcerative colitis. In certain embodiments, the inflammatory disorder or disease is psoriasis. In certain embodiments, the inflammatory disorder or disease is lupus. In certain embodiments, the inflammatory disorder or disease is rheumatoid arthritis.

[0225] In certain aspects, described herein is a method of treating a mammalian subject in need of treatment of a medical condition associated with an elevated level of IL-13, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody described herein or a pharmaceutical composition described herein.

[0226] In certain aspects, described herein is a method of reducing the bioactivity of IL-13 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.

[0227] In certain aspects, described herein is a method of inhibiting a type 2 helper T (Th2)-mediated allergic reaction 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.

[0228] In certain aspects, described herein is a method of decreasing the level of thymus and activation-regulated chemokine (TARC) / CCL17 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.

[0229] In certain aspects, described herein is a method of preventing an inflammatory disorder 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.

[0230] These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description and the accompanying drawings.

Brief Description of the Drawings

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DETAILED DESCRIPTION OF THE INVENTION

[0246] IL-13 signal transduction begins with the binding of IL-13 to IL-13Rα1, forming an inactive complex, which then binds to IL-4Rα to form a fully active receptor heterodimer. This active receptor heterodimer is involved in the pathogenesis of atopic dermatitis. The present disclosure relates, in part, to anti-IL-13 antibodies that prevent the formation of this heterodimer.

[0247] As shown in Figure 1, in the 3D rendering of human IL-13, the gray color of "1" highlights the epitope of lebrikizumab, which overlaps with the epitope of a specific antibody disclosed herein. Importantly, these epitopes also overlap with the IL-4Rα epitope on IL-13. Without wishing to be bound by theory, it is believed that antibodies that bind to this region are likely to prevent the formation of the IL-13Rα1-IL-4Rα heterodimer and limit the inflammatory signaling leading to atopic dermatitis. In contrast, the epitope of tralokinumab-ldrm (Adbry™) highlighted in gray of "3" does not overlap with the IL-4Rα epitope on IL-13 and thus may have further limited ability to prevent heterodimerization.

[0248] Definitions Unless otherwise defined, all technical terms, notations, and other scientific terms used herein shall have the meanings commonly understood by those of ordinary skill in the art. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a difference beyond what is generally understood in the art. The techniques and procedures described or referred to herein are generally well understood and are commonly utilized by those of ordinary skill in the art using conventional methods, such as the widely used molecular cloning methods described in Sambrook et al., Molecular Cloning: A Laboratory Manual 4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Procedures involving the use of commercially available kits and reagents are generally, unless otherwise specified, carried out according to the protocols and conditions defined by the manufacturer.

[0249] As used herein, the singular forms "a", "an", and "the" include plural references unless otherwise indicated.

[0250] It is understood that the aspects and embodiments of the invention described herein include those “comprising,” “consisting of,” and “consisting essentially of” the aspects and embodiments.

[0251] For all compositions described herein and all methods of using the compositions described herein, the compositions may, in some cases, include the recited ingredients or steps, or may “consist essentially of” the recited ingredients or steps. When a composition is described as “consisting essentially of” the recited ingredients, the composition includes the recited ingredients and may include other ingredients that do not substantially affect the condition being treated, but does not include any other ingredients that substantially affect the condition being treated other than those explicitly recited, or, if the composition includes additional ingredients that substantially affect the condition being treated other than those recited, the composition does not include the additional ingredients in a concentration or amount sufficient to substantially affect the condition being treated. When a method is described as “consisting essentially of” the recited steps, the method includes the recited steps and may include other steps that do not substantially affect the condition being treated, but the method does not include any other steps that substantially affect the condition being treated other than those explicitly recited. By way of non-limiting specific example, when a composition is described as “consisting essentially of” a certain ingredient, the composition may further include any amount of a pharmaceutically acceptable carrier, medium, or diluent and other ingredients that do not substantially affect the condition being treated.

[0252] As used herein, the term “vector” refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is ligated. The term includes vectors as self-replicating nucleic acid structures and vectors that are integrated into the genome of a host cell into which they are introduced. A particular vector is capable of expressing a nucleic acid to which it is operably linked. Such vectors are referred to herein as “expression vectors.”

[0253] The terms "host cell", "host cell line", and "host cell culture" are used interchangeably and refer to a cell into which an exogenous nucleic acid has been introduced and the progeny of such a cell. A host cell includes "transformants" (or "transformed cells") and "transfectants" (or "transfected cells"), each of which includes the primary transformed or transfected cell and progeny derived therefrom. Such progeny may not be identical, in nucleic acid content, to the parental cell and may include mutations. A "recombinant host cell" or "host cell" refers to a cell containing an exogenous polynucleotide, regardless of the method used for insertion, e.g., direct uptake, transduction, f mating, or other methods known in the art for creating recombinant host cells.

[0254] As used herein, the term "eukaryote" refers to an organism belonging to the phylogenetic domain Eucarya, such as, but not limited to, animals (including, but not limited to, mammals, insects, reptiles, birds, etc.), ciliates, plants (including, but not limited to, monocots, dicots, algae, etc.), fungi, yeast, flagellates, microsporidia, protists, etc.

[0255] As used herein, the term "prokaryote" refers to a prokaryotic organism. For example, a prokaryote may belong to the eubacteria phylogenetic domain (including, but not limited to, Escherichia coli, Thermus thermophilus, Bacillus stearothermophilus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, etc.) or the archaebacteria phylogenetic domain (including, but not limited to, Methanococcus jannaschii, Methanobacterium thermoautotrophicum, Halobacterium, such as Haloferax volcanii and Halobacterium species NRC-1, Archaeoglobus fulgidus, Pyrococcus furiosus, Pyrococcus horikoshii, Aeuropyrum pernix, etc.).

[0256] As used herein, "effective amount" or "therapeutically effective amount" refers to the amount of a therapeutic compound, e.g., an anti-IL-13 antibody, that is effective, either alone or in combination with another therapy, in bringing about or contributing to a desired therapeutic effect when administered to an individual, either as a single dose or as part of a series of doses. Examples of desired therapeutic effects include enhancing an immune response, slowing or delaying tumorigenesis, stabilizing a disease, and improving one or more symptoms. An effective amount may be administered in one or more dosages.

[0257] The term "treating" (and its variations such as "treat" or "treatment") refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject who requires such treatment. Treatment may be performed in the course of clinical pathology. Desirable effects of treatment include preventing recurrence of a disease, alleviating symptoms, reducing any direct or indirect pathological consequences of a disease, preventing metastasis, reducing the rate of disease progression, improving or alleviating the medical condition, and improving remission or prognosis.

[0258] The term "sufficient amount" means an amount sufficient to achieve the desired effect, e.g., an amount sufficient to modulate the immune response in a subject.

[0259] As used herein, the terms "subject" or "individual" mean a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. In some embodiments, the subject has a disease or condition that can be treated with an antibody provided herein. In some embodiments, the disease or disorder is cancer. In some embodiments, the disease or condition is a viral infection.

[0260] The term "in vitro" refers to processes occurring in living cells grown separately from a living organism, e.g., growth in tissue culture.

[0261] The term "in vivo" refers to processes occurring in a living organism.

[0262] The term "package insert" is used to refer to the instructions for use, which customarily include information about indications, uses, dosages, administrations, combination therapies, contraindications, and / or warnings regarding the use of such therapeutic or diagnostic agents, included in the commercial package (e.g., kit) of a therapeutic or diagnostic agent.

[0263] The term "pharmaceutical composition" refers to a preparation in which the biological activity of the active ingredient contained therein is in a form effective for treatment of a subject and which does not contain additional ingredients that are unacceptably harmful to the subject in the amounts provided in the pharmaceutical composition.

[0264] The terms "co-administer", "co-administering", and "in combination with" include administering two or more therapeutic agents simultaneously, concurrently, or sequentially, without a specific time limit. In one embodiment, the agents are present intracellularly or in the subject's body simultaneously, or exert biological or therapeutic effects simultaneously. In one embodiment, the therapeutic agents are included in the same composition or unit dosage form. In other embodiments, the therapeutic agents are included in separate compositions or unit dosage forms. In certain embodiments, the first agent can be administered prior to administration of the second therapeutic agent.

[0265] The terms "modulate" and "modulation" refer to decreasing or suppressing the recited variable element, or alternatively, activating or increasing it.

[0266] The terms "increase" and "activate" refer to an increase in the recited variable element 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.

[0267] The terms "decrease" and "suppress" refer to a decrease in the recited variable element of 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.

[0268] The term "about" indicates and encompasses the indicated value and the range above and below that value. In certain embodiments, the term "about" indicates the specified value ±10%, ±5%, or ±1%. In certain embodiments, where applicable, the term "about" indicates the specified value(s) ± one standard deviation of that value(s).

[0269] The term "stimulate" refers to the activation of receptor signaling to induce a biological response associated with the activation of a receptor. An "agonist" is an entity that binds to a receptor and stimulates it.

[0270] The term "antagonize" refers to the inhibition of receptor signaling to suppress a biological response associated with the activation of a receptor. An "antagonist" is an entity that binds to a receptor and antagonizes it.

[0271] For any of the structural and functional characteristics described herein, methods for measuring these characteristics are known in the art.

[0272] The term "optionally", when used consecutively, means including any one to all of the recited combinations and contemplates all sub - combinations.

[0273] The term "amino acid" refers to the 20 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), glutamine (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).

[0274] The term "affinity" refers to the total strength of 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 the intrinsic binding affinity, which reflects a 1:1 interaction between the members of the binding pair (e.g., an antibody and an antigen or epitope).

[0275] As used herein, the term "kd" (min⁻¹) refers to the dissociation rate constant of a particular antibody-antigen interaction. This value is also referred to as the koff value.

[0276] As used herein, the term "ka" (M⁻¹×min⁻¹) refers to the association rate constant of a particular antibody-antigen interaction. This value is also referred to as the kon value.

[0277] As used herein, the term "KD" (M) refers to the dissociation equilibrium constant of a particular antibody-antigen interaction. KD = kd / ka. In some embodiments, the affinity of an antibody is expressed in terms of KD for the interaction between such antibody and its antigen. For clarity, as is known in the art, a lower KD value indicates a stronger affinity interaction, and a higher KD value indicates a weaker affinity interaction.

[0278] As used herein, the term "KA" (M⁻¹) refers to the association equilibrium constant of a particular antibody-antigen interaction. KA = ka / kd.

[0279] The term "antibody" is used herein in its broadest sense and includes certain types of immunoglobulin molecules that contain one or more antigen-binding domains that specifically bind to an antigen or epitope. Antibodies include, in particular, intact antibodies (e.g., intact immunoglobulins), antibody fragments, and multispecific antibodies.

[0280] An "anti-IL-13 antibody", "IL-13 antibody", or "IL-13-specific antibody" is an antibody provided herein that specifically binds to the antigen IL-13.

[0281] The term "epitope" means the portion of an antigen that specifically binds to an antibody.

[0282] As used herein, the terms "hypervariable region" or "HVR" each refer to regions of an antibody variable domain that are hypervariable within an array and / or form structurally defined loops ("hypervariable loops").

[0283] The term "antigen-binding domain" means the part of an antibody that is capable of specifically binding to an antigen or epitope.

[0284] The term "chimeric antibody" refers to an antibody in which portions of the heavy and / or light chains are derived from a particular origin or species, while the remaining portions of the heavy and / or light chains are derived from a different origin or species.

[0285] The term "human antibody" refers to an antibody produced by a human or human cell, or an antibody having an amino acid sequence corresponding to that of an antibody of non-human origin that uses a human antibody repertoire or a sequence encoding a human antibody (e.g., obtained from or newly designed from a human origin). Human antibodies strictly exclude humanized antibodies.

[0286] The term "humanized antibody" refers to a protein having a sequence different from that of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and / or additions such that, when the humanized antibody is administered to a human subject, it is less likely to induce an immune response and / or induces a less severe immune response compared to an antibody of a non-human species.

[0287] The term "bispecific antibody" means an antibody that contains two or more different antigen-binding domains that collectively specifically bind to two or more different epitopes.

[0288] A "monospecific antibody" is an antibody that contains one or more binding sites that specifically bind to a single epitope. Examples of monospecific antibodies are naturally occurring IgG molecules that are bivalent (i.e., have two antigen-binding domains) and recognize the same epitope with each of the two antigen-binding domains. The binding specificity can exist in any suitable valence.

[0289] The term "monoclonal antibody" refers to an antibody from a population of substantially identical antibodies. A population of substantially identical antibodies is substantially similar, excluding variants that may normally arise during the production of monoclonal antibodies, and includes antibodies that bind to the same epitope(s). Such variants generally exist only in small amounts. Monoclonal antibodies are typically obtained by a process that involves the selection of a single antibody from a plurality of antibodies. For example, the selection process can be the selection of a unique clone from a pool of a plurality of clones, such as hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones. The selected antibody can be further modified, for example, to improve its affinity for the target ("affinity maturation"), to humanize the antibody, to improve its production in cell culture, and / or to reduce its immunogenicity in a subject.

[0290] The term "single-chain" refers to a molecule that includes amino acid monomers linearly linked by peptide bonds. In certain such embodiments, the C-terminus of the Fab light chain is connected to the N-terminus of the Fab heavy chain in a single-chain Fab molecule. As described in more detail herein, an scFv has a variable domain of the light chain (VL) connected by a polypeptide chain to the N-terminus of the variable domain of the heavy chain (VH) from its C-terminus. Alternatively, the scFv includes a polypeptide chain, and the C-terminus of the VH is connected by the polypeptide chain to the N-terminus of the VL.

[0291] A "Fab fragment" (also called an antigen-binding fragment) includes the variable domains VL and VH of the light and heavy chains, respectively, together with the constant domain of the light chain (CL) and the first constant domain of the heavy chain (CH1). The variable domains include complementarity-determining loops (CDRs, also called hypervariable regions) that are involved in antigen binding. A Fab' fragment differs from a Fab fragment in that it has the addition of several residues at the carboxy terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region.

[0292] The "F(ab’)2" fragment contains two Fab’ fragments linked by disulfide bonds near the hinge region. The F(ab’)2 fragment can be generated, for example, by recombinant methods or by pepsin digestion of intact antibodies. The F(ab’) fragment can be dissociated, for example, by treatment with β-mercaptoethanol.

[0293] The "Fv" fragment contains a dimer of one heavy chain variable domain and one light chain variable domain non-covalently linked.

[0294] "Single-chain Fv" or "sFv" or "scFv" contains the VH and VL domains of an antibody, and these domains are present in a single polypeptide chain. In one embodiment, the Fv polypeptide further contains a polypeptide linker between the VH and VL domains, whereby the scFv can form a structure desirable for antigen binding. For a review of scFv, see, for example, Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenberg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994). The scFv fragments of HER2 antibodies are described in WO93 / 16185, U.S. Patent No. 5,571,894, and U.S. Patent No. 5,587,458.

[0295] The "scFv-Fc" fragment contains an scFv bound to an Fc domain. For example, the Fc domain can be bound to the C-terminus of the scFv. The Fc domain can follow VH or VL depending on the orientation of the variable domains of 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 contains the Fc domain of IgG4.

[0296] 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 the other variable domain. Single domain antibodies, and fragments thereof, are described in Arabi Ghahroudi et al., FEBS Letters, 1998, 414:521-526 and Muyldermans et al., Trends in Biochem. Sci., 2001, 26:230-245, each of which is incorporated herein by reference in its entirety. Single domain antibodies are also known as sdAbs or nanobodies. SdAbs are quite stable and are easily expressed as fusion partners with the Fc chain of an antibody (Harmsen MM, De Haard HJ (2007). “Properties, production, and applications of camelid single-domain antibody fragments”. Appl. Microbiol Biotechnol. 77(1):13-22).

[0297] The terms "full-length antibody", "intact antibody", and "whole antibody" are used interchangeably herein and refer to an antibody having a structure with a heavy chain that includes an Fc region that is substantially similar to a naturally occurring antibody structure. For example, when used to refer to an IgG molecule, a "full-length antibody" is an antibody that includes two heavy chains and two light chains.

[0298] The term "antibody fragment" refers to a portion of an intact antibody, e.g., an antibody that includes an antigen-binding region or variable region of an intact antibody. Examples of antibody fragments include, for example, Fv fragments, Fab fragments, F(ab’)2 fragments, Fab’ fragments, scFv (sFv) fragments, and scFv-Fc fragments.

[0299] The term "Fc domain" or "Fc region" as used herein is used to define the C-terminal region of an immunoglobulin heavy chain that includes at least a portion of the constant region. The term includes the Fc region of a native sequence and variant Fc regions.

[0300] The term "substantially purified" means that the constructs described herein, or variants thereof, may substantially or essentially lack the proteins normally associated with or interacting with them in their natural environment, i.e., in the case of a recombinantly produced antibody substantially free of cellular material, the host cell has 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% (dry weight) of contaminating protein.

[0301] The term "percent identity" in the context of two or more nucleic acid or polypeptide sequences refers to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., publicly available computer software such as BLAST, BLASTP, BLASTN, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software or algorithms available to those of skill in the art) or by visual inspection. Software for performing BLAST analyses is generally available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). Those of skill in the art can determine appropriate parameters for aligning sequences, including any algorithms required to obtain the maximum alignment over the full length of the sequences being compared. Depending on the application, the "percent identity" may exist over a region of the sequences being compared, e.g., over a functional domain, or alternatively, over the full length of the two sequences being compared.

[0302] For array comparison, usually, one array functions as a reference array against which a test array is compared. When using an array comparison algorithm, the test array and the reference array are input into a computer, and if necessary, partial array coordinates are specified and array algorithm program parameters are specified. Thereafter, the array comparison algorithm calculates the percentage of array identity of the test array(s) relative to the reference array based on the specified program parameters.

[0303] Optimal alignment of the arrays for comparison can be performed by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the similarity search method of Pearson & Lipman, Proc. Nat’l. Acad. Sci. USA 85:2444 (1988), computer implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA, Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or visual inspection (generally Ausubel et al., see below).

[0304] It is understood that the ranges recited herein are shorthand for all values within the ranges including the recited endpoints. For example, the range of 1 to 50 is understood to include any number, combination of numbers, or subrange from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50.

[0305] It should be noted that, as used in this specification and the appended claims, the singular forms "a", "an", and "the" include the plural referents unless the context clearly dictates otherwise.

[0306] Anti-IL-13 antibody Structure of the antibody This application provides antibodies that bind to IL-13 and compositions comprising the antibodies.

[0307] Well-known immunoglobulin genes include kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as numerous immunoglobulin variable region genes. Light chains are classified into either kappa or lambda. The "class" of an antibody or immunoglobulin refers to the type of constant domain or constant region that its heavy chain has. There are five major classes of antibodies, namely, IgA, IgD, IgE, IgG, and IgM, and some of these can be further classified into subclasses (isotypes), for example, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0308] The structural unit of an exemplary immunoglobulin (antibody) is composed of two pairs of polypeptide chains, each pair having one "light" chain (about 25 kD) and one "heavy" chain (about 50 - 70 kD). The N-terminal domain of each chain defines a variable region of about 100 - 110 or more amino acids that is mainly involved in antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light chain domains and heavy chain domains, respectively. The heavy chain of IgG1 contains VH, CH1, CH2, and CH3 domains, respectively, from the N-terminus to the C-terminus. The light chain contains the VL domain and the CL domain from the N-terminus to the C-terminus. The heavy chain of IgG1 contains a hinge between the CH1 domain and the CH2 domain. In certain embodiments, the immunoglobulin construct comprises at least one immunoglobulin domain from IgG, IgM, IgA, IgD, or IgE connected to a therapeutic polypeptide. In some embodiments, the immunoglobulin domains found in the antibodies provided herein are from, or derived from, immunoglobulin-based constructs such as diabodies or nanobodies. In certain embodiments, the immunoglobulin constructs described herein comprise at least one immunoglobulin domain from heavy chain antibodies such as camelid antibodies. In certain embodiments, the immunoglobulin constructs provided herein comprise at least one immunoglobulin domain from mammalian antibodies such as bovine antibodies, human antibodies, camelid antibodies, mouse antibodies, or any chimeric antibody.

[0309] 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.

[0310] 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.

[0311] Generally, a native four-chain antibody contains six hypervariable regions (HVRs), i.e., three in VH (H1, H2, and H3) as well as three in VL (L1, L2, and L3). HVRs generally contain amino acid residues from hypervariable loops and / or from complementarity-determining regions (CDRs), the latter being those where sequence variability is highest and / or which are involved in antigen recognition. Except for CDR1 in VH, CDRs generally contain the amino acid residues that form the hypervariable loops. HVRs are also called CDRs, and these terms are used interchangeably herein to refer to the portions of the variable regions that form the antigen-binding domain. This particular region has been described by Kabat et al., U.S. Dept. of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and Chothia et al., J Mol Biol 196:901-917 (1987), and their definitions, when compared to each other, include overlaps or subsets of amino acid residues. Nevertheless, the application of any definition for referring to the CDRs of an antibody or its variant is intended to be within the scope of the terms defined and used herein. The exact residue numbers encompassing a particular CDR vary depending on the sequence and size of that CDR. Given the amino acid sequence of the variable region of an antibody, one of ordinary skill in the art can routinely determine which residues are included in a particular CDR.

[0312] The boundaries of the CDR amino acid sequences can be determined by one of several known numbering schemes, including those described in Kabat et al., supra (the "Kabat" numbering scheme), Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (the "Chothia" numbering scheme), MacCallum et al., 1996, J. Mol. Biol. 262:732-745 (the "Contact" numbering scheme), Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 (the "IMGT" numbering scheme), and Honegge and Pluckthun, J. Mol. Biol., 2001, 309:657-70 (the "AHo" numbering scheme), each of which is incorporated by reference in its entirety.

[0313] Table 1 provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes. For CDR-H1, the residues are numbered using both the Kabat and Chothia numbering schemes.

[0314] The CDRs may be assigned, for example, using antibody numbering software, such as Abnum described in Abhinandan and Martin, Immunology, 2008, 45:3832-3839, which is incorporated by reference in its entirety and is available at, for example, www.bioinf.org.uk / abs / abnum / . [Table 1]

[0315] The "EU numbering scheme" is generally used when referring to the residues of the antibody heavy chain constant region (e.g., as reported by Kabat et al., supra). Unless otherwise indicated, the EU numbering scheme is used to refer to the residues of the antibody heavy chain constant region described herein.

[0316] One example of an antigen-binding domain is the antigen-binding domain formed by the VH-VL dimer of an antibody. Another example of an antigen-binding domain is the antigen-binding domain formed by diversification of a particular loop from the 10th fibronectin type III domain of an adnectin. An antigen-binding domain can include CDR1, 2, and 3 from the heavy chain, in this order, and CDR1, 2, and 3 from the light chain, in this order.

[0317] An epitope often consists of surface-accessible amino acid residues and / or sugar side chains and can have specific three-dimensional structural characteristics and specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that in the presence of a denaturing solvent, binding to the former may be lost, but binding to the latter may not be lost. An epitope can include amino acid residues directly involved in binding and other amino acid residues not directly involved in binding. The epitope to which an antibody binds can be identified using known techniques for determining epitopes, such as testing the binding of an antibody to IL-13 variants having different point mutations or chimeric IL-13 variants.

[0318] To screen for antibodies that bind to an epitope on a target antigen (e.g., IL-13) to which the antibody of interest binds, a conventional cross-blocking assay, such as those described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed. Alternatively, or additionally, epitope mapping can be performed by methods known in the art.

[0319] A chimeric antibody is an antibody in which a portion of the heavy chain and / or light chain is derived from a specific origin or species, while the remaining portion of the heavy chain and / or light chain is derived from a different origin or species.

[0320] A human antibody is an antibody produced by a human or human cell, or an antibody having an amino acid sequence corresponding to that of an antibody of non-human origin using a human antibody repertoire or a sequence encoding a human antibody (e.g., obtained from or newly designed from a human origin). A human antibody strictly excludes humanized antibodies.

[0321] A humanized antibody has a sequence that is different 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 the humanized antibody is administered to a human subject, it is less likely to induce an immune response and / or is less likely to induce an immune response that is as severe as compared to a non-human species antibody. In one embodiment, certain amino acids within the framework and constant domains of the heavy chain and / or light chain of a non-human species antibody are mutated to produce a humanized antibody. In another embodiment, a constant domain(s) from a human antibody is fused to a variable domain(s) from a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are altered such that the potential immunogenicity of the non-human antibody when administered to a human subject is decreased. In this case, the altered amino acid residues are such that the change to the amino acid sequence is a conservative change such that it is not critical for its immunospecific binding to the antigen of the antibody or such that the binding of the humanized antibody to the antigen is not significantly inferior to the binding of the non-humanized antibody to the antigen. Examples of methods for making humanized antibodies can be found in U.S. Pat. Nos. 6,054,297, 5,886,152, and 5,877,293. For further details, see Jones et al., Nature, 1986, 321:522-525, Riechmann et al., Nature, 1988, 332:323-329, and Presta, Curr. Struct. Biol., 1992, 2:593-596. Each of these is incorporated by reference in its entirety.

[0322] Two or more different epitopes may be epitopes on the same antigen (e.g., a single IL-13) or on different antigens (e.g., different IL-13 molecules, or an IL-13 molecule and a non-IL-13 molecule). In some embodiments, a multispecific antibody binds to two different epitopes (i.e., a "bispecific antibody"). In some embodiments, a multispecific antibody binds to three different epitopes (i.e., a "trispecific antibody").

[0323] Examples of anti-IL-13 antibodies include those described herein, such as the clones described in the drawings and / or tables. In some embodiments, the antibody comprises an alternative backbone. In some embodiments, the antibody consists of an alternative backbone. In some embodiments, the antibody consists essentially of an alternative backbone. In some embodiments, the antibody comprises an antibody fragment. In some embodiments, the antibody consists of an antibody fragment. In some embodiments, the antibody consists essentially of an antibody fragment.

[0324] In some embodiments, the antibody is a monoclonal antibody.

[0325] In some embodiments, the antibody is a polyclonal antibody.

[0326] In some embodiments, the antibody is produced by a hybridoma. In other embodiments, the antibody is produced by recombinant cells engineered to express the desired variable and constant domains.

[0327] In some embodiments, the antibody may be a single-chain antibody or other antibody derivatives or variants thereof that retain antigen specificity and a lower hinge region.

[0328] In some embodiments, the antibody can be a multifunctional antibody, a recombinant antibody, a human antibody, a humanized antibody, fragments or variants thereof. In certain embodiments, the antibody fragment or derivative thereof is selected from Fab fragment, Fab’2 fragment, CDR, and ScFv.

[0329] In some embodiments, the antibody is capable of forming an immune complex. For example, the immune complex can be a tumor cell coated with the antibody.

[0330] For array comparison, typically, one array functions as a reference array against which a test array is compared. When using an array comparison algorithm, the test array and the reference array are input into a computer, and if necessary, partial array coordinates are specified and array algorithm program parameters are specified. Thereafter, the array comparison algorithm calculates the percent sequence identity of the test array(s) relative to the reference array based on the specified program parameters.

[0331] Optimal alignment of the sequences for comparison can be conducted by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the similarity search method of Pearson & Lipman, Proc. Nat’l. Acad. Sci. USA 85:2444 (1988), computer implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA, the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (generally see Ausubel et al., see below).

[0332] An example of an algorithm suitable for measuring percent sequence identity and percent sequence similarity is the BLAST algorithm, which is described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analysis is generally available through the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov / ).

[0333] Sequence of the IL-13 antibody VH domain In some embodiments, the antibodies provided herein comprise a VH sequence selected from SEQ ID NOs: 1-32 and 470.

[0334] In some embodiments, the antibodies provided herein include VH sequences having at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the exemplary VH sequences set forth in SEQ ID NOs: 1-32 and 470. In some embodiments, the antibodies provided herein include VH sequences having up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid substitutions to the V H sequences set forth in SEQ ID NOs: 1-32 and 470. In some embodiments, 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 obtained 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 may, for example, be newly isolated according to the methods provided herein for obtaining antibodies.

[0335] VL domain In some embodiments, the antibodies provided herein include VL sequences selected from SEQ ID NOs: 33-57 and 471.

[0336] In some embodiments, the antibodies provided herein include VL sequences having at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the exemplary VL sequences set forth in SEQ ID NOs: 33-57 and 471. In some embodiments, the antibodies provided herein include VL sequences set forth in SEQ ID NOs: 33-57 and 471 having up to 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 embodiments, 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 obtained 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 may, for example, be newly isolated according to the methods provided herein for obtaining antibodies.

[0337] VH-VL combination In some embodiments, the antibodies provided herein include VH sequences selected from SEQ ID NOs: 1-32 and 470, and VL sequences selected from SEQ ID NOs: 33-57 and 471, for example, VH-VL combinations as set forth in Table 2 below.

[0338] In certain aspects, any one of SEQ ID NOs: 1-32 and 470 can be combined with any one of SEQ ID NOs: 33-57 and 471.

[0339] In certain embodiments, the antibody includes a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence set forth in SEQ ID NO: 49.

[0340] In certain embodiments, the antibody comprises a VH sequence selected from the sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence set forth in SEQ ID NO: 51.

[0341] In some embodiments, the antibodies provided herein have a VH sequence having at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the exemplary VH sequences set forth in SEQ ID NOs: 1-32 and 470, and a VL sequence having at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the exemplary VL sequences set forth in SEQ ID NOs: 33-57 and 471. In some embodiments, the antibodies provided herein comprise a VH sequence set forth in SEQ ID NOs: 1-32 and 470 having up to 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 set forth in SEQ ID NOs: 33-57 and 471 having up to 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 embodiments, 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 obtained 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 may, for example, be newly isolated according to the methods provided herein for obtaining antibodies.

[0342] In some embodiments, the antibodies provided herein include VH and VL sequences selected from the combinations set forth in Table 2 below. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 1 and the VL sequence set forth in SEQ ID NO: 33. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 2 and the VL sequence set forth in SEQ ID NO: 33. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 35. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 35. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 35. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 35. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 35. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 36. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 36. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 36. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 36. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 36. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody includes the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 40.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 40. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 40. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 40. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 40. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 42. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 9 and the VL sequence set forth in SEQ ID NO: 43. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 44. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 45. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 46. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 47. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 48. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 49. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 50. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 52. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 53. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 54. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 55.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 56. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 57. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 10 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 11 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 12 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 13 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436 - 468 and 484 - 539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 14 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 16 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 17 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 18 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 19 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 20 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 21 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 22 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 23 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 24 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 25 and the VL sequence set forth in SEQ ID NO: 39.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 26 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 27 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 29 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 30 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 31 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 32 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471.

[0343] In certain embodiments, the isolated antibody comprises a heavy chain variable domain comprising a framework region sequence selected from the sequences set forth in SEQ ID NOs: 198 - 229, 255 - 256, 258 - 259, 261 - 285, 311 - 315, 317 - 342, 368 - 369, 371 - 399, and 540 - 580. In certain embodiments, the isolated antibody comprises a heavy chain variable domain comprising 1, 2, 3, or 4 framework region sequences selected from the sequences set forth in SEQ ID NOs: 198 - 229, 255 - 256, 258 - 259, 261 - 285, 311 - 315, 317 - 342, 368 - 369, 371 - 399, and 540 - 580.

[0344] In certain embodiments, the isolated antibody comprises a light chain variable domain comprising a framework region sequence selected from the sequences set forth in SEQ ID NOs: 230-231, 233-235, 239, 241-254, 286, 288, 290-291, 293, 296-310, 343-345, 347, 400-424, and 581-609. In certain embodiments, the isolated antibody comprises a light chain variable domain comprising one, two, three, or four framework region sequences selected from the sequences set forth in SEQ ID NOs: 230-231, 233-235, 239, 241-254, 286, 288, 290-291, 293, 296-310, 343-345, 347, 400-424, and 581-609.

[0345] In certain embodiments, the isolated antibody comprises a heavy chain variable domain comprising one, two, three, or four framework region sequences selected from the sequences set forth in SEQ ID NOs: 198-229, 255-256, 258-259, 261-285, 311-315, 317-342, 368-369, 371-399, and 540-580, and a light chain variable domain comprising one, two, three, or four framework region sequences selected from the sequences set forth in SEQ ID NOs: 230-231, 233-235, 239, 241-254, 286, 288, 290-291, 293, 296-310, 343-345, 347, 400-424, and 581-609.

Table 2-1

Table 2-2

Table 2-3

Table 2-4

Table 2-5

Table 2-6

Table 2-7

Table 2-8

Table 2-9

Table 2-10

Table 2-11

Table 2-12

Table 2-13

Table 2-14

Table 2-15

Table 2-16

[0346] In some embodiments, the HC constant domain of such IgG4-SP has the following sequence: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 427).

[0347] In some embodiments, the HC constant domain of such hIgG1-LALA-YTE has the following sequence: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 439).

[0348] In some embodiments, the HC constant domain of such hIgG1-LAGA YTE has the following sequence: ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELAGAPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 440).

[0349] In some embodiments, the HC constant domain of such hIgG1-LALA-LS has the following sequence:

[0350] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVLHEALHSHYTQKSLSLSPG (SEQ ID NO: 446).

[0351] In some embodiments, the HC constant domain of such IgG4-YTE has the following sequence: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 457).

[0352] In some embodiments, the HC constant domain of such IgG4-LS has the following sequence: ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHSYTQKSLSLSLGK (SEQ ID NO: 460).

[0353] In some embodiments, the human kappa LC constant domain has the following sequence: RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 469). CDR

[0354] In some embodiments, the antibodies provided herein include one to three CDRs of a VH domain selected from SEQ ID NOs: 1-32 and 470, for example, any of the CDRs listed in Table 3, Table 4, or Table 5 below. In some embodiments, the antibodies provided herein include two to three CDRs of a VH domain selected from SEQ ID NOs: 1-32 and 470. In some embodiments, the antibodies provided herein include three CDRs of a VH domain selected from SEQ ID NOs: 1-32 and 470. In some embodiments, the CDRs are exemplary CDRs. In some embodiments, the CDRs are Kabat CDRs. In some embodiments, the CDRs are Chothia CDRs. In some embodiments, the CDRs are IMGT CDRs. In some embodiments, the CDRs are AbM CDRs. In some embodiments, the CDRs are Contact CDRs.

[0355] 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-H1, CDR-H2, or CDR-H3 of SEQ ID NOs: 58-140. In some embodiments, the CDR-H1 is the CDR-H1 of a VH domain selected from SEQ ID NOs: 1-32 and 470 having at most 1, 2, 3, 4, or 5 amino acid substitutions. In some embodiments, the CDR-H2 is the CDR-H2 of the VH domain of SEQ ID NOs: 1-32 and 470 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, the CDR-H3 is the CDR-H3 of a VH domain selected from SEQ ID NOs: 1-32 and 470 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0356] In some embodiments, the antibodies provided herein comprise one to three CDRs of the VL domains of SEQ ID NOs: 33-57 and 471, for example, any of the CDRs listed in Table 6, Table 7, or Table 8 below. In some embodiments, the antibodies provided herein comprise two to three CDRs of the VL domains of SEQ ID NOs: 33-57 and 471. In some embodiments, the antibodies provided herein comprise three CDRs of the VL domains of SEQ ID NOs: 33-57 and 471. In some embodiments, the CDRs are exemplary CDRs. In some embodiments, the CDRs are Kabat CDRs. In some embodiments, the CDRs are Chothia CDRs. In some embodiments, the CDRs are IMGT CDRs. In some embodiments, the CDRs are AbM CDRs. In some embodiments, the CDRs are Contact CDRs.

[0357] 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 with the CDR-L1, CDR-L2, or CDR-L3 of SEQ ID NOs: 141-188. In some embodiments, the CDR-L1 is the CDR-L1 of the VL domain of SEQ ID NOs: 33-57 and 471 having 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: 33-57 and 471 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, the CDR-L3 is the CDR-L3 of the VL domain of SEQ ID NOs: 33-57 and 471 having at most 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0358] In some embodiments, the antibodies provided herein comprise 1 to 3 CDRs of the VH domain selected from SEQ ID NOs: 1-32 and 470, and 1 to 3 CDRs of the VL domain of SEQ ID NOs: 33-57 and 471. In some embodiments, the antibodies provided herein comprise 2 to 3 CDRs of the VH domain selected from SEQ ID NOs: 1-32 and 470, and 2 to 3 CDRs of the VL domain of SEQ ID NOs: 33-57 and 471. In some embodiments, the antibodies provided herein comprise 3 CDRs of the VH domain selected from SEQ ID NOs: 1-32 and 470, and 3 CDRs of the VL domain of SEQ ID NOs: 33-57 and 471. In some embodiments, the CDRs are exemplary CDRs. In some embodiments, the CDRs are Kabat CDRs. In some embodiments, the CDRs are Chothia CDRs. In some embodiments, the CDRs are IMGT CDRs. In some embodiments, the CDRs are AbM CDRs. In some embodiments, the CDRs are Contact CDRs.

[0359] In some embodiments, the antibodies provided herein comprise a CDR-H3 selected from SEQ ID NOs: 112-120 and 130-40. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-H3 of SEQ ID NOs: 112-120 or 130-40. In some embodiments, the CDR-H3 is a CDR-H3 selected from SEQ ID NOs: 112-120 and 130-40 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0360] In some embodiments, the antibodies provided herein include CDR-H1s of SEQ ID NOs: 58-99 and 121. In some embodiments, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H1 of SEQ ID NOs: 58-99 or 121. In some embodiments, the CDR-H1 is a CDR-H1 of SEQ ID NOs: 58-99 or 121 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0361] In some embodiments, the antibodies provided herein include any one of the CDR-H2s of SEQ ID NOs: 100-111. In some embodiments, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with any one of the CDR-H2s of SEQ ID NOs: 100-111. In some embodiments, the CDR-H2 is any one of the CDR-H2s of SEQ ID NOs: 100-111 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may, for example, be newly isolated according to the methods provided herein for obtaining antibodies.

[0362] In some embodiments, the antibodies provided herein comprise a CDR-L3 selected from SEQ ID NOs: 165-172. In some embodiments, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-L3 of SEQ ID NOs: 165-172. In some embodiments, the CDR-L3 is a CDR-L3 of SEQ ID NOs: 165-172 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0363] In some embodiments, the antibodies provided herein include a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS. In some embodiments, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS. In some embodiments, the CDR-L2 is a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0364] In some embodiments, the antibodies provided herein comprise a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152. In some embodiments, the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152. In some embodiments, the CDR-L1 is a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, 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 obtained 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 may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0365] In some embodiments, the antibodies provided herein comprise a CDR-H3 selected from SEQ ID NOs: 112-120 and 130-140, a CDR-H2 of SEQ ID NOs: 100-111, a CDR-H1 selected from SEQ ID NOs: 58-99 and 121, a CDR-L3 selected from SEQ ID NOs: 165-172, a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS, and a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-H3 selected from SEQ ID NOs: 112-120 and 130-140, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-H2 of SEQ ID NOs: 100-111, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-H1 selected from SEQ ID NOs: 58-99 and 121, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-L3 selected from SEQ ID NOs: 165-172, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152.In some embodiments, the CDR-H3 is a CDR-H3 selected from SEQ ID NOs: 112-120 and 130-140 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NOs: 100-111 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 selected from SEQ ID NOs: 58-99 and 121 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 selected from SEQ ID NOs: 165-172 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 selected from SEQ ID NOs: 153-158 and the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 selected from SEQ ID NOs: 141-144 and 149-152 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0366] In some embodiments, the antibodies provided herein include the CDR-H3 of SEQ ID NO: 112, 121, and 130, the CDR-H2 of SEQ ID NO: 100, 104, and 108, the CDR-H1 of SEQ ID NO: 58, 68, and 85, the CDR-L3 of SEQ ID NO: 168, 173, and 181, the CDR-L2 of SEQ ID NO: 153 and the amino acid sequence LAS, and the CDR-L1 of SEQ ID NO: 141 and 149. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H1 of SEQ ID NO: 58, 68, or 85, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L3 of SEQ ID NO: 168, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L1 of SEQ ID NO: 141 or 149.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 112 or 130 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 100, 104, or 108 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 58, 68, or 85 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 168 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is the CDR-L1 of SEQ ID NO: 141 or 149 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0367] In some embodiments, the antibodies provided herein comprise a CDR-H3 of SEQ ID NO: 112, 121, or 130, a CDR-H2 of SEQ ID NO: 100, 104, or 108, a CDR-H1 of SEQ ID NO: 58, 68, or 85, a CDR-L3 of SEQ ID NO: 165, a CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS, and a CDR-L1 of SEQ ID NO: 141 or 149. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-H1 of SEQ ID NO: 58, 68, or 85, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-L3 of SEQ ID NO: 165, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the CDR-L1 of SEQ ID NO: 141 or 149.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 112 or 130 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 100, 104, or 108 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 58, 68, or 85 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 165 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; the CDR-L1 is the CDR-L1 of SEQ ID NO: 141 or 149 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0368] In some embodiments, the antibodies provided herein comprise CDR-H3 of SEQ ID NO: 112 or 130, CDR-H2 of SEQ ID NO: 100, 104, or 108, CDR-H1 of SEQ ID NO: 58, 68, or 85, CDR-L3 of SEQ ID NO: 165, CDR-L2 of SEQ ID NO: 158 or amino acid sequence LAS, and CDR-L1 of SEQ ID NO: 141 or 149. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H1 of SEQ ID NO: 58, 68, or 85, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L3 of SEQ ID NO: 165, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L2 of SEQ ID NO: 158 or amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L1 of SEQ ID NO: 141 or 149.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 112 or 130 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 100, 104, or 108 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 58, 68, or 85 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 165 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 158 or the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; the CDR-L1 is the CDR-L1 of SEQ ID NO: 141 or 149 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0369] In some embodiments, the antibodies provided herein include CDR-H3 of SEQ ID NO: 112, 121, or 130, CDR-H2 of SEQ ID NO: 100, 104, or 108, CDR-H1 of SEQ ID NO: 58, 67, or 84, CDR-L3 of SEQ ID NO: 165, CDR-L2 of SEQ ID NO: 153 or amino acid sequence LAS, and CDR-L1 of SEQ ID NO: 141 or 149. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H1 of SEQ ID NO: 58, 67, or 84, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L3 of SEQ ID NO: 165, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L2 of SEQ ID NO: 153 or amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L1 of SEQ ID NO: 141 or 149.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 112 or 130 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 100, 104, or 108 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 58, 67, or 84 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 165 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 153 or the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; the CDR-L1 is the CDR-L1 of SEQ ID NO: 141 or 149 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0370] In some embodiments, the antibodies provided herein include the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 of SEQ ID NO: 58, 67, or 84, the CDR-L3 of SEQ ID NO: 165, the CDR-L2 of SEQ ID NO: 158 or the amino acid sequence LAS, and the CDR-L1 of SEQ ID NO: 141 or 149. In some embodiments, the CDR-H3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H3 of SEQ ID NO: 112 or 130, the CDR-H2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H2 of SEQ ID NO: 100, 104, or 108, the CDR-H1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-H1 of SEQ ID NO: 58, 67, or 84, the CDR-L3 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L3 of SEQ ID NO: 165, the CDR-L2 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L2 of SEQ ID NO: 158 or the amino acid sequence LAS, and the CDR-L1 has at least about 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity with the CDR-L1 of SEQ ID NO: 141 or 149.In some embodiments, the CDR-H3 is the CDR-H3 of SEQ ID NO: 112 or 130 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is the CDR-H2 of SEQ ID NO: 100, 104, or 108 having up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is the CDR-H1 of SEQ ID NO: 58, 67, or 84 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is the CDR-L3 of SEQ ID NO: 165 having up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is the CDR-L2 of SEQ ID NO: 158 or the amino acid sequence LAS having up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is the CDR-L1 of SEQ ID NO: 141 or 149 having up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.

[0371] In some embodiments, the amino acid substitutions are conservative amino acid substitutions. 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 obtained 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 or clones are not derived from the sequences provided herein and may be newly isolated, for example, according to the methods provided herein for obtaining antibodies.

[0372] In certain aspects, the antibodies disclosed herein do not include the antibodies disclosed in U.S. Patent No. 9,067,994.

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Table 4-5

Table 4-6

Table 4-7

Table 4-8

Table 4-9

Table 4-10

Table 4-11

Table 4-12

Table 5-1

Table 5-2

Table 5-3

Table 5-4

Table 5-5

Table 5-6

Table 5-7

Table 5-8

Table 5-9

Table 5-10

Table 5-11

Table 6-1

Table 6-2

Table 6-3

Table 6-4

Table 6-5

Table 6-6

Table 6-7

Table 7-1

Table 7-2

Table 7-3

Table 7-4

Table 7-5

Table 7-6

Table 7-7

Table 8-1

Table 8-2

Table 8-3

Table 8-4

Table 8-5

Table 8-6

Table 8-7

Table 8-8

Table 8-9

[0373] Fc region The structures of the Fc regions of various immunoglobulins and the glycosylation sites contained therein are known in the art. See Schroeder and Cavacini, J. Allergy Clin. Immunol., 2010, 125: S41-52, which is incorporated herein by reference in its entirety. The Fc region may be a naturally occurring Fc region or an Fc region modified as known in the art or described elsewhere in the present disclosure.

[0374] Unless otherwise specified herein, the numbering of amino acid residues within the Fc region or constant region follows the EU numbering system, also known as the EU index, 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, the "Fc polypeptide" of a dimeric Fc refers to one of the two polypeptides that form the dimeric Fc domain, i.e., a polypeptide containing the C-terminal constant region of an immunoglobulin heavy chain capable of stable self-association. For example, the Fc polypeptide of the Fc of dimeric IgG contains the constant domain sequences of CH2 of IgG and CH3 of IgG. Fc can be of the classes IgA, IgD, IgE, IgG, and IgM, and some of these can be further classified into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0375] The terms "Fc receptor" and "FcR" are used to describe receptors that bind to the Fc region of an antibody. For example, an FcR can be a native sequence human FcR. In general, FcRs are those that bind to IgG antibodies (gamma receptors) and include receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic polymorphisms and forms due to alternative splicing of these receptors. FcγRII receptors include FcγRIIA ("activating receptor") and FcγRIIB ("inhibitory receptor"), which have similar amino acid sequences that mainly differ in their cytoplasmic domains. Other isotypes of immunoglobulins can also be bound by certain FcRs (see, e.g., Janeway et al., Immuno Biology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999)). The activating receptor FcγRIIA contains an immunoreceptor activation tyrosine motif (ITAM) in its cytoplasmic domain. The inhibitory receptor FcγRIIB contains an immunoreceptor inhibitory tyrosine motif (ITIM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991), Capel et al., Immunomethods 4:25-34 (1994), and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). 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 involved in the transfer of maternal IgG to the fetus (Guyer et al., J. Immunol. 117:587 (1976), and Kim et al., J. Immunol. 24:249 (1994)).

[0376] Modifications in the CH2 domain can affect the binding of FcR to Fc. To selectively alter the affinity of Fc for different Fc gamma receptors, several amino acid modifications in the Fc region are known in the art. In some embodiments, the Fc comprises one or more modifications to facilitate selective binding of Fc-gamma receptors.

[0377] Exemplary mutations that alter the binding of FcR to Fc are listed below:

[0378] S298A / E333A / K334A, S298A / E333A / K334A / K326A (Lu Y, Vernes JM, Chiang N, et al. J Immunol Methods. 2011 Feb 28;365(1-2):132-41),

[0379] F243L / R292P / Y300L / V305I / P396L, F243L / R292P / Y300L / L235V / P396L (Stavenhagen JB, Gorlatov S, Tuaillon N, et al. Cancer Res. 2007 Sep 15;67(18):8882-90; Nordstrom JL, Gorlatov S, Zhang W, et al. Breast Cancer Res. 2011 Nov 30;13(6):R123),

[0380] F243L (Stewart R, Thom G, Levens M, et al. Protein Eng Des Sel. 2011 Sep;24(9):671-8.), S298A / E333A / K334A (Shields RL, Namenuk AK, Hong K, et al. J Biol Chem. 2001 Mar 2;276(9):6591-604),

[0381] S239D / I332E / A330L, S239D / I332E (Lazar GA, Dang W, Karki S, et al. Proc Natl Acad Sci USA. 2006 Mar 14;103(11):4005-10),

[0382] S239D / S267E, S267E / L328F (Chu SY, Vostiar I, Karki S, et al. Mol Immunol. 2008 Sep;45(15):3926-33), and

[0383] 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 described 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, Oct 2012) describes mutations on page 283.

[0384] 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 defucosylation, or engineering of the affinity of the Fc for activating receptors, namely FCGR3a for ADCC and C1q for CDC. Table 9 below summarizes various designs reported in the literature for engineering effector functions.

[0385] Methods for producing antibodies with little or no fucose at the Fc glycosylation site (Asn297, EU numbering) without changing the amino acid sequence are well known in the art. The GlymaX® technology (ProBioGen AG) is based on the introduction of genes of enzymes that divert the cellular pathway of fucose biosynthesis into the cells used for antibody production. This prevents the addition of the sugar "fucose" to the carbohydrate moiety of the N-linked antibody by the antibody-producing cells (von Horsten et al. (2010) Glycobiology. 2010 Dec;20(12):1607-18). Another approach for obtaining antibodies with reduced fucosylation levels can be found in U.S. Patent No. 8,409,572, which teaches that cell lines for antibody production can be selected for their ability to reduce the level of fucosylation of the antibody, such that they can be fully defucosylated (meaning undetectable fucose is present) or that they can be partially defucosylated, i.e., 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 similar antibodies produced by mammalian expression systems.

[0386] Thus, in one embodiment, the antibodies described herein can include a dimeric Fc comprising one or more amino acid modifications set forth in Table 9 that confer improved effector function. In another embodiment, the antibody can be defucosylated to improve effector function.

Table 9

[0387] Fc modifications that reduce the binding and / or effector function of FcγR and / or complement are known in the art. Recent publications have described strategies used to engineer antibodies with reduced or silenced 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” In Therapeutic Antibody Engineering, Cambridge: Woodhead Publishing (2012), pp 225-249). These strategies include reducing effector function through modification of glycosylation, use of the IgG2 / IgG4 backbone, or introduction of mutations in the hinge or CH2 region of Fc. For example, US Patent Publication No. 2011 / 0212087 (Strohl), International Patent Publication No. WO2006 / 105338 (Xencor), US Patent Publication No. 2012 / 0225058 (Xencor), US Patent Publication No. 2012 / 0251531 (Genentech), and Strop et al. ((2012) J. Mol. Biol. 420:204-219) describe specific modifications for reducing the binding of FcγR or complement to Fc.

[0388] Specific non-limiting examples of known amino acid modifications for reducing the binding of FcγR or complement to Fc are identified in Table 10 below.

Table 10

[0389] Examples of cell lines capable of producing defucosylated antibodies include CHO-DG44 that stably overexpresses the bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4-hexulose reductase (RMD) (see Henning von Horsten et al., Glycobiol 2010, 20:1607-1618), or Lec13 CHO cells deficient in protein fucosylation (see Ripka et al., Arch. Biochem. Biophys., 1986, 249:533-545, U.S. Patent Publication No. 2003 / 0157108, WO 2004 / 056312. Each of these is incorporated by reference in its entirety), and knockout cell lines, such as alpha-1,6-fucosyltransferase gene or FUT8 knockout CHO cells (see Yamane-Ohnuki et al., Biotech. Bioeng., 2004, 87:614-622, Kanda et al., Biotechnol. Bioeng., 2006, 94:680-688, and WO 2003 / 085107. Each of these is incorporated by reference in its entirety). Another approach for obtaining antibodies with reduced levels of fucosylation can be found in U.S. Patent No. 8,409,572, which teaches selecting cell lines for antibody production for their ability to reduce the level of fucosylation of the antibody.

[0390] Antibodies can be fully defucosylated (meaning they contain no detectable fucose), or they can be partially defucosylated, i.e., 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 similar antibodies produced by mammalian expression systems.

[0391] In some embodiments, the antibodies provided herein include an IgG1 domain having a reduced fucose content at position Asn297 as compared to a naturally occurring IgG1 domain. Such Fc domains are known to have improved ADCC. See Shields et al., J. Biol. Chem., 2002, 277:26733-26740, which is incorporated by reference in its entirety. In some embodiments, such antibodies are devoid of fucose at position Asn297. The amount of fucose can be measured using any suitable method, such as the method described in WO2008 / 077546, which is incorporated by reference in its entirety.

[0392] In certain embodiments, the antibodies provided herein include an Fc region having one or more amino acid substitutions that improve ADCC, for example, having substitutions at one or more of positions 298, 333, and 334 of the Fc region. In some embodiments, the antibodies provided herein include an Fc region having one or more amino acid substitutions at positions 239, 332, and 330, as described in Lazar et al., Proc. Natl. Acad. Sci. USA, 2006, 103:4005-4010, which is incorporated by reference in its entirety.

[0393] Other exemplary glycosylation variants that can be incorporated into the antibodies provided herein are described, for example, in U.S. Patent Publications Nos. 2003 / 0157108, 2004 / 0093621, 2003 / 0157108, 2003 / 0115614, 2002 / 0164328, 2004 / 0093621, 2004 / 0132140, 2004 / 0110704, 2004 / 0110282, 2004 / 0109865, International Patent Publications Nos. 2000 / 61739, 2001 / 29246, 2003 / 085119, 2003 / 084570, 2005 / 035586, 2005 / 035778, 2005 / 053742, 2002 / 031140, Okazaki et al., J. Mol. Biol., 2004, 336:1239-1249, and Yamane-Ohnuki et al., Biotech. Bioeng., 2004, 87:614-622. Each of these is incorporated by reference in its entirety.

[0394] In some embodiments, the antibodies provided herein comprise an Fc region having at least one galactose residue on the oligosaccharide attached to the Fc region. Such antibody variants can have improved CDC function. Examples of such antibody variants are described, for example, in WO1997 / 30087, WO1998 / 58964, and WO1999 / 22764, each of which is incorporated by reference in its entirety.

[0395] In some embodiments, the antibodies provided herein comprise one or more modifications that improve or decrease C1q binding and / or CDC. See U.S. Patent No. 6,194,551, WO99 / 51642, and Idusogie et al., J. Immunol., 2000, 164:4178-4184. Each of these is incorporated by reference in its entirety.

[0396] In certain embodiments, the antibodies provided herein comprise a heavy chain comprising a constant heavy chain sequence selected from the sequences set forth in SEQ ID NOs: 425-468 and 484-539.

[0397] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 1 and the VL sequence set forth in SEQ ID NO: 33, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 2 and the VL sequence set forth in SEQ ID NO: 33, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 35, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 36, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 439, 440, 446, 457, and 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NO: 436-468. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 4 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 5 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 6 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 40, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 42, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 9 and the VL sequence set forth in SEQ ID NO: 43, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 44, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 45, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 46, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 47, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 48, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 49, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 50, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 52, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 53, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 54, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 55, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 56, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 57, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 10 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 11 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 12 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 13 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 14 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 16 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 17 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 18 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 19 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the arrange. Comprising the VH sequence described in column number 20 and the VL sequence described in SEQ ID NO: 39, wherein the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 21 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 22 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 23 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 24 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 25 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 26 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 27 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence described in SEQ ID NO: 28 and the VL sequence described in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences described in SEQ ID NOs: 436-468 and 484-539.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 28 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 29 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 30 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 31 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 32 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequences set forth in SEQ ID NOs: 436-468 and 484-539.

[0398] In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 15 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 8 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises the human IgG sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 39, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 3 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446.In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7 and the VL sequence set forth in SEQ ID NO: 51, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 439. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 446. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 470 and the VL sequence set forth in SEQ ID NO: 471, and the human Fc region comprises a human IgG sequence selected from the sequence set forth in SEQ ID NO: 460.

[0399] In certain embodiments, the isolated antibody described herein comprises the constant light chain sequence represented by SEQ ID NO: 469.

[0400] In certain embodiments, the Fc region comprises one or more amino acid substitutions, which result in an extended half-life of the antibody, increased ADCC activity, increased ADCP activity, or increased CDC activity as compared to an Fc that does not contain the one or more substitutions. In certain embodiments, the one or more amino acid substitutions result in an extended half-life of the antibody at pH 6.0 as compared to an antibody comprising a wild-type Fc region. In certain embodiments, the isolated antibody comprising an Fc region having the one or more amino acid substitutions has a half-life of about 80 to 110 days in humans.

[0401] In certain embodiments, the antibody has an extended half-life that 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 an antibody comprising a wild-type Fc region. In certain embodiments, the antibody has an extended half-life that 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 rituximab.

[0402] In certain embodiments, the Fc region comprises one or more amino acid substitutions that result in an extended half-life of the antibody compared to an Fc that does not comprise the one or more substitutions and a decrease in one or more of ADCC activity, ADCP activity, or CDC activity. In certain embodiments, the one or more amino acid substitutions result in an extended half-life of the antibody at pH 6.0 compared to an antibody comprising a wild-type Fc region. In certain embodiments, an isolated antibody comprising an Fc region having the one or more amino acid substitutions has a half-life in humans of about 80 to 110 days.

[0403] 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 are 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 (LA), M428L / N434AComprising a plurality of amino acid substitutions selected from the group consisting of L234A / G237A (LALA), L234A / L235A / G237A, L234A / L235A / P329G, N297A, D265A / YTE, 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, 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, LALAGA / YD, LALAPG / YD, T307Q / Q311V / A378V (QVV), N297A / QVV, D265A / QVV, LALA / QVV, LAGA / QVV, LALAGA / QVV, LALAPG / QVV, DDRVV, N297A / DDRVV, D265A / DDRVV, LALA / DDRVV, LAGA / DDRVV, LALAGA / DDRVV, and LALAPG / DDRVV.

[0404] In certain embodiments, the one or more amino acid substitutions are selected from the group consisting of LALA / YTE, LAGA / YTE, LALA / LS, YTE, and LS.

[0405] In certain embodiments, the one or more amino acid substitutions comprise or consist of LALA / YTE. In certain embodiments, the one or more amino acid substitutions comprise or consist of LAGA / YTE. In certain embodiments, the one or more amino acid substitutions comprise or consist of LALA / LS. In certain embodiments, the one or more amino acid substitutions comprise or consist of YTE. In certain embodiments, the one or more amino acid substitutions comprise or consist of LS.

[0406]

[0407] In certain embodiments, the Fc region binds to an Fcγ receptor selected from the group consisting of FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, and FcγRIIIb. In certain embodiments, the Fc region binds to an Fcγ receptor with a higher affinity compared to an antibody comprising a wild-type Fc region at pH 6.0.

[0408] Binding The affinity of molecule X for its partner Y can be expressed by the dissociation equilibrium constant (K D ). The kinetic elements contributing to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).

[0409] Regarding the binding of an antibody to a target molecule, the terms "bind to", "specifically bind", "specifically binds to", "specific", "selectively binds to", and "selective" with respect to a particular antigen (e.g., a polypeptide target) or an epitope of a particular antigen mean binding that is clearly distinct from non-specific or non-selective interactions (e.g., with non-target molecules). Specific binding can be measured, for example, by measuring the binding to the target molecule (i.e., IL-13) and comparing it to the binding to non-target molecules. Specific binding can also be measured by competition with a control molecule that mimics the epitope recognized on the target molecule. In that case, specific binding is indicated if the binding of the antibody to the target molecule is competitively inhibited by the control molecule. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 50% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 40% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 30% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 20% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 10% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 1% of the affinity for IL-13. In some embodiments, the affinity of the anti-IL-13 antibody for a non-target molecule is less than about 0.1% of the affinity for IL-13.

[0410] As used herein in the context of two or more antibodies, the terms “compete with” or “cross-compete with” indicate that the two or more antibodies compete for binding to an antigen (e.g., IL-13). In one exemplary assay, IL-13 is coated on a surface and contacted with a primary anti-IL-13 antibody, followed by addition of a secondary anti-IL-13 antibody. In another exemplary assay, a primary anti-IL-13 antibody is coated on a surface, contacted with IL-13, followed by addition of a secondary anti-IL-13 antibody. If the presence of the primary anti-IL-13 antibody reduces the binding of the secondary anti-IL-13 antibody, in either assay, these antibodies compete with each other. The term “compete with” includes combinations of antibodies where one antibody reduces the binding of another, but no competition is observed when these antibodies are added in the reverse order. However, in some embodiments, the primary and secondary antibodies inhibit each other's binding regardless of the order in which they are added. In some embodiments, one antibody reduces the binding of another antibody to its antigen, as measured in a competitive binding assay, 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%. One of ordinary skill in the art can select the concentration of antibodies used in a competitive assay based on the affinity of the antibodies for IL-13 and the valency of the antibodies. The assays described in this definition are exemplary, and one of ordinary skill in the art can use any suitable assay to determine whether antibodies compete with each other. Suitable assays are described, for example, in Cox et al., “Immunoassay Methods,” in Assay Guidance Manual [Internet], updated December 24, 2014 (ncbi.nlm.nih.gov / books / NBK92434 / , accessed September 29, 2015), Silman et al., Cytometry, 2001, 44:30-37, and Finco et al., J. Pharm. Biomed. Anal., 2011, 54:351-358. Each of these is incorporated by reference in its entirety.

[0411] A test antibody competes with a reference antibody when an excess of the test antibody (e.g., at least 2-fold, 5-fold, 10-fold, 20-fold, or 100-fold) inhibits or blocks the binding of the reference antibody, as measured in a competitive binding assay, by, for example, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. Antibodies identified by a competitive assay (competitive antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes that are proximal enough to cause steric hindrance to the epitope to which the reference antibody binds. For example, a secondary competitive antibody that competes with respect to the binding of a primary antibody described herein to IL-13 can be identified. In some cases, the secondary antibody can block or inhibit the binding of the primary antibody, as measured in a competitive binding assay, by, for example, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. In some cases, the secondary antibody can displace the primary antibody by more than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%.

[0412] In certain embodiments, the antibody binds to human IL-13.

[0413] In certain embodiments, the antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475.

[0414] In certain embodiments, the antibody is cross-reactive with cynomolgus IL-13.

[0415] In certain embodiments, the antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475, as measured by SPR, with a K D of about 1, 2, 3, 4, 5, 6, 7, 8, 9 x 10 -9 M or less. In certain embodiments, the antibody binds to the IL-13 sequences set forth in SEQ ID NOs: 472-475, as measured by SPR, with a K D of about 1 x 10 -10Bind at less than M. In certain embodiments, the antibody binds to human IL-13 with a K D of about 1x10 -9 or less.

[0416] In some embodiments, the antibodies provided herein bind to IL-13 with a K D of about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10x10 -8 or less, as measured by ELISA or any other suitable method known in the art. In some embodiments, the antibodies provided herein bind to IL-13 with a K D of about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10x10 -9 or less.

[0417] In some embodiments, the K D of the antibodies provided herein for binding to IL-13 is about 0.001 - 0.01, 0.01 - 0.1, 0.01 - 0.05, 0.05 - 0.1, 0.1 - 0.5, 0.5 - 1, 0.25 - 0.75, 0.25 - 0.5, 0.5 - 0.75, 0.75 - 1, 0.75 - 2, 1.1 - 1.2, 1.2 - 1.3, 1.3 - 1.4, 1.4 - 1.5, 1.5 - 1.6, 1.6 - 1.7, 1.7 - 1.8, 1.8 - 1.9, 1.9 - 2, 1 - 2, 1 - 5, 2 - 7, 3 - 8, 3 - 5, 4 - 6, 5 - 7, 6 - 8, 7 - 9, 7 - 10, or 5 - 10x10, as measured by ELISA or any other suitable method known in the art.-8 It is M. In some embodiments, the antibodies provided herein bind to IL-13 with a K D of about 1x10 -8 M or less, or about 1x10 -9 M or less.

[0418] In some embodiments, the antibodies provided herein bind to IL-13 with a K D of about 10, 9, 8, 7, 6, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.98, 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, 1, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.05, 0.01, 0.005, 0.001, 0.0005, or 0.0001x10 -8 M or less, or less than that. In some embodiments, the antibodies provided herein bind to IL-13 with a K D of 5 - 3, 4 - 2, 3 - 1, 1.9 - 1.8, 1.8 - 1.7, 1.7 - 1.6, 1.6 - 1.5, 1.9 - 1.5, 1.5 - 1, 1 - 0.8, 1 - 0.5, 0.9 - 0.6, 0.7 - 0.4, 0.6 - 0.2, 0.5 - 0.3, 0.3 - 0.2, 0.2 - 0.1, 0.1 - 0.01, 0.01 - 0.001, or 0.001 - 0.0001×10 -8 M.

[0419] Function "Effector function" refers to a biological activity mediated by the Fc region of an antibody, which may vary depending on the isotype of the antibody. Examples of the effector functions of an antibody include blocking, agonizing, or antagonizing a receptor ligand, C1q binding for activating complement-dependent cytotoxicity (CDC), Fc receptor binding for activating antibody-dependent cell-mediated cytotoxicity (ADCC), and antibody-dependent cell phagocytosis (ADCP). In some embodiments, the effector function of the anti-IL-13 antibody described herein is antagonism, which blocks the binding of the IL-13 receptor to IL-13.

[0420] Pharmaceutical composition This application provides compositions comprising an antibody, including pharmaceutical compositions comprising any one or more of the antibodies described herein together with one or more pharmaceutically acceptable excipients. In some embodiments, the composition is sterile. The pharmaceutical composition generally comprises an effective amount of the antibody.

[0421] These compositions can include, in addition to one or more of the antibodies disclosed herein, 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 effectiveness of the active ingredient. The detailed properties of the carrier or other materials can depend on the route of administration, e.g., oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes.

[0422] Pharmaceutical compositions for oral administration can be in the form of tablets, capsules, powders or liquids. Tablets can contain solid carriers, e.g., gelatin or adjuvants. Liquid pharmaceutical compositions generally contain liquid carriers, e.g., water, petroleum, animal or vegetable oils, mineral oil or synthetic oils. They may also contain physiological saline, dextrose or other sugar solutions or glycols, e.g., ethylene glycol, propylene glycol or polyethylene glycol.

[0423] In the case of intravenous injection, cutaneous injection or subcutaneous injection, or injection at the affected part, the active ingredient is in the form of a parenterally acceptable aqueous solution that is pyrogen-free and has an appropriate pH, isotonicity and stability. Those skilled in the art can readily prepare an appropriate solution, for example, using an isotonic medium such as sodium chloride injection solution, Ringer's injection solution, lactated Ringer's injection solution. If necessary, preservatives, stabilizers, buffers, antioxidants and / or other additives may be included.

[0424] Administration of the anti-IL-13 antibody to an individual is preferably a "therapeutically effective amount" or a "prophylactically effective amount" (in some cases, prophylaxis can be considered as treatment), which is sufficient to be effective against the individual. The actual dosage administered, as well as the rate and time course of administration, depend on the nature and severity of the protein aggregation disease being treated. The determination of the treatment regimen, such as dosage, etc., is the responsibility of the general practitioner and other physicians and usually takes into account the disorder being treated, the condition of the individual patient, the site of delivery, the method of administration, and other factors known to the physician. Examples of the above techniques and protocols can be found in Remington’s Pharmaceutical Sciences, 16 th edition, Osol, A. (ed), 1980.

[0425] The composition can be administered alone or in combination with other treatments, simultaneously or sequentially, depending on the condition being treated.

[0426] Method Preparation method 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. Such nucleic acid can encode an amino acid sequence comprising the VL of the antibody and / or an amino acid sequence comprising the VH (e.g., the light and / or heavy chains of the antibody), or an amino acid sequence comprising the VHH of a single domain antibody. In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acid are provided. In one embodiment, the nucleic acid is provided in a multicistronic vector. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, the host cell comprises (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 an 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 (e.g., is transformed thereby). In one embodiment, the host cell is eukaryotic, e.g., Chinese hamster ovary (CHO) cells or human embryonic kidney (HEK) cells, or lymphoid cells (e.g., Y0, NS0, Sp20 cells). In one embodiment, a method for making an antibody is provided, the method comprising culturing a host cell comprising a nucleic acid encoding the antibody described above under conditions suitable for expression of the antibody, and optionally, recovering the antibody from the host cell (or the host cell's medium).

[0427] With respect to recombinant production of antibodies, for example, a nucleic acid encoding the antibody described above is isolated and inserted into one or more vectors for further cloning and / or expression in a host cell. Such nucleic acid can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that specifically bind to genes encoding the heavy and light chains of the antibody).

[0428] When an antibody or variant thereof is recombinantly produced by a host cell, the protein is, in certain embodiments, 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 cell. When an antibody or variant thereof is recombinantly produced by a host cell, the protein is, in certain embodiments, present in the 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 cell. In certain embodiments, a "substantially purified" antibody produced by the methods described herein has 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%, particularly at least about 75%, 80%, 85%, more specifically at least about 90%, at least about 95%, at least about 99% or more, as measured by suitable methods such as SDS / PAGE analysis, RP-HPLC, SEC, and capillary electrophoresis.

[0429] Suitable host cells for cloning or expressing a vector encoding an antibody include the prokaryotic or eukaryotic cells described herein.

[0430] A recombinant host cell or host cell is a cell containing an exogenous polynucleotide, regardless of the method used for insertion, such as direct uptake, transduction, f-mating, or other methods known in the art for creating recombinant host cells. The exogenous polynucleotide may be maintained as a non-integrating vector, such as a plasmid, or alternatively, may be integrated into the host genome. Host cells can include CHO, derivatives of CHO, NS0, Sp2O, CV-1, VERO-76, HeLa, HepG2, Per.C6, or BHK.

[0431] For example, antibodies can be produced in bacteria, particularly when glycosylation and Fc effector functions are not required. For the expression of antibody fragments and polypeptides in bacteria, see, for example, U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, which describes the expression of antibody fragments in E. coli.). After expression, the antibody may be isolated from the soluble fraction of the bacterial cell paste and further purified.

[0432] In addition to prokaryotes, eukaryotic microorganisms, such as filamentous fungi or yeast strains, including fungi and yeast strains whose glycosylation pathways have been "humanized" to result in the production of antibodies with partial or complete human glycosylation patterns, are suitable cloning or expression hosts for vectors encoding antibodies. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).

[0433] Host cells suitable for the expression of glycosylated antibodies also originate from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. In conjunction with insect cells, a number of baculovirus strains have been identified that can be used, particularly for the transfection of Spodoptera frugiperda cells.

[0434] Plant cell cultures can also be used as hosts. See, for example, U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429, which describe the PLANTIBODIES™ technology for producing antibodies in transgenic plants.

[0435] Vertebrate cells can also be used as hosts. For example, animal cell lines adapted to grow in suspension are also useful. Other examples of useful mammalian host cell lines include monkey kidney CV1 cells transformed by SV40 (COS-7), human fetal kidney cell lines (e.g., 293 or 293 cells described in Graham et al., J. Gen Virol. 36:59 (1977)), baby hamster kidney cells (BHK), mouse Sertoli cells (e.g., TM4 cells described in Mather, Biol. Reprod. 23:243-251 (1980)), monkey kidney cells (CV1), African green monkey kidney cells (VERO-76), human cervical cancer cells (HELA), dog kidney cells (MDCK), buffalo rat hepatocytes (BRL 3A), human lung cells (W138), human hepatocytes (Hep G2), mouse mammary tumor (MMT 060562), TRI cells (e.g., described in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)), MRC5 cells, and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells including DHFR-CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)), as well as 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 (B.K.C. Lo, ed., Humana Press, Totowa, N.J.), pp. 255-268 (2003).

[0436] In one embodiment, the antibodies described herein are produced by a method comprising transfecting at least one stable mammalian cell with a nucleic acid encoding the antibody at a predetermined ratio in a stable mammalian cell and expressing the nucleic acid in the at least one mammalian cell. In some embodiments, the predetermined ratio of the nucleic acid is determined by a transient transfection experiment to determine the relative ratio of the input nucleic acid that results in the highest percentage of antibody in the expression product.

[0437] In some embodiments, in the method for producing an antibody in a stable mammalian cell described herein, the expression product of the at least one stable mammalian cell comprises a high percentage of the desired glycosylated antibody compared to the monomeric heavy or light chain polypeptide, or other antibodies.

[0438] In some embodiments, in the method for producing a glycosylated antibody in a stable mammalian cell described herein, identifying and purifying the desired glycosylated antibody is included. In some embodiments, the identification is by one or both of liquid chromatography and mass spectrometry.

[0439] Optionally, the antibody can be purified or isolated after expression. Proteins can be isolated or purified by a variety of methods known to those of skill in the art. Standard purification methods include chromatographic techniques such as ion exchange, hydrophobic interaction, affinity, sizing, or gel filtration, and reverse phase, performed at atmospheric or high pressure using systems such as FPLC and HPLC. Also included as purification methods are electrophoretic techniques, immunological techniques, precipitation techniques, dialysis techniques, and chromatofocusing techniques. Ultrafiltration and diafiltration techniques combined with protein concentration are also useful. As is well known in the art, various natural proteins bind to Fc and antibodies, and these proteins can be used in the present invention for antibody purification. For example, bacterial protein A and G bind to the Fc region. Similarly, bacterial protein L binds to the Fab region of some antibodies. Purification can often be made possible by a specific fusion partner. For example, an antibody can be purified using glutathione resin when a GST fusion is used, and Ni when a His tag is used. +2It can be purified using affinity chromatography or, if a flag tag is used, using an immobilized anti-flag antibody. For general guidance on appropriate purification techniques, see, for example, Protein Purification: Principles and Practice, 3 rd Ed., Scopes, Springer-Verlag, NY, 1994, which is incorporated by reference in its entirety. The degree of purification required varies depending on the use of the antibody. In some cases, purification is not necessary.

[0440] In certain embodiments, the antibody is purified using anion exchange chromatography including, but not limited to, chromatography on Q-Sepharose, DEAE-Sepharose, Poros HQ, Poros DEAF, Toyopearl Q, Toyopearl QAE, Toypearl DEAE, Resource / Source Q, and DEAE, Fractogel Q and DEAE columns.

[0441] In certain embodiments, the proteins described herein are purified using cation exchange chromatography including, but not limited to, chromatography on SP-Sepharose, CM-Sepharose, Poros HS, Poros CM, Toyopearl SP, Toyopearl CM, Resource / Source S and CM, Fractogel S and CM columns and their equivalents and comparators.

[0442] Furthermore, the antibodies described herein can be chemically synthesized using techniques known in the art (see, e.g., Creighton, 1983, Proteins: Structures and Molecular Principles, W.H. Freeman & Co., N.Y and Hunkapiller et al., Nature, 310:105-111 (1984)). For example, polypeptides corresponding to fragments of a polypeptide can be synthesized using a peptide synthesizer. Additionally, non-classical amino acids or chemical amino acid analogs can be introduced as substitutions or additions to the polypeptide sequence as needed. Non-classical amino acids generally include, but are not limited to, D-isomers of common amino acids, 2,4-diaminobutyric acid, alpha-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, cysteic acid, 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. Further, the amino acids can be either D (right-handed) or L (left-handed).

[0443] In certain embodiments, the antibodies described herein have an aggregation temperature greater than about 69°C, greater than about 70°C, greater than about 71°C, greater than about 72°C, greater than about 73°C, greater than about 74°C, greater than about 75°C, or greater than about 76°C, e.g., from about 69°C to about 77°C, from about 70°C to about 76°C, from about 71°C to about 75°C. In certain embodiments, the aggregation temperature is measured using DSF.

[0444] In certain embodiments, the antibodies described herein have a lower hydrophobicity compared to rituximab as measured by hydrophobic interaction chromatography (HIC). In certain embodiments, the antibody exhibits an HIC retention time of less than about 15.2 minutes. In certain embodiments, the antibody exhibits an HIC retention time of from about 13 minutes to about 15 minutes.

[0445] Method of Use In one aspect, the present application provides a method of contacting IL-13 with an anti-IL-13 antibody, e.g., a human or humanized antibody, thereby resulting in inhibition of the binding of IL-13 to the IL-13 receptor expressed on cells.

[0446] In one aspect, the present application provides a method of using the isolated anti-IL-13 antibodies described herein for the treatment of a disorder or disease in a subject. In certain embodiments, described herein is a method of treating a subject in need of treatment with an anti-IL-13 antibody, the method comprising administering to a mammalian subject a therapeutically effective amount of an anti-IL-13 antibody described herein or a pharmaceutical composition comprising an anti-IL-13 antibody. In certain specific embodiments, the present application provides a method of treating a disorder or disease associated with an elevated level of IL-13 and / or IgE in a subject.

[0447] In certain embodiments, described herein is a method of treating a medical condition associated with IL-13 activity, the method comprising administering to a mammalian subject a therapeutically effective amount of an isolated anti-IL-13 antibody described herein or a pharmaceutical composition comprising an isolated anti-IL-13 antibody.

[0448] In certain embodiments, described herein is a method of treating a mammalian subject in need of treatment for an inflammatory disorder or disease, 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 disorder or disease is atopic dermatitis. In certain embodiments, the inflammatory disorder or disease is asthma. In certain embodiments, the inflammatory disorder or disease is idiopathic pulmonary fibrosis. In certain embodiments of the methods described herein, the inflammatory disorder or disease is alopecia areata. In certain embodiments, the inflammatory disorder or disease is chronic rhinosinusitis with nasal polyps. In certain embodiments, the inflammatory disorder or disease is chronic rhinosinusitis without nasal polyps (CRSsNP). In certain embodiments, the inflammatory disorder or disease is eosinophilic esophagitis (EoE). In certain embodiments, 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). In certain embodiments, the inflammatory disorder or disease is Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA). In certain embodiments, the inflammatory disorder or disease is prurigo nodularis (PN). In certain embodiments, the inflammatory disorder or disease is chronic spontaneous urticaria (CSU). In certain embodiments, the inflammatory disorder or disease is chronic pruritus of unknown origin (CPUO). In certain embodiments, the inflammatory disorder or disease is bullous pemphigoid (BP). In certain embodiments, the inflammatory disorder or disease is cold urticaria (ColdU). In certain embodiments, the inflammatory disorder or disease is allergic fungal rhinosinusitis (AFRS). In certain embodiments, the inflammatory disorder or disease is allergic bronchopulmonary aspergillosis (ABPA). In certain embodiments, the inflammatory disorder or disease is chronic obstructive pulmonary disease (COPD).In certain embodiments, the inflammatory disorder or disease is an inflammatory bowel disease, such as Crohn's disease or ulcerative colitis. In certain embodiments, the inflammatory disorder or disease is psoriasis. In certain embodiments, the inflammatory disorder or disease is lupus. In certain embodiments, the inflammatory disorder or disease is rheumatoid arthritis.

[0449] In certain aspects, described herein is a method of treating a mammalian subject in need of treatment of a medical condition associated with an elevated level of IL-13, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0450] In certain aspects, described herein is a method of decreasing the biological activity of IL-13 in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0451] In certain aspects, described herein is a method of inhibiting a type 2 helper T (Th2)-mediated allergic response in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0452] In certain aspects, described herein is a method of inhibiting IL-13-induced phosphorylation of signal transducer and activator of transcription 6 (STAT6) in a cell, the method comprising contacting the cell with an antibody described herein.

[0453] In certain aspects, described herein is a method of inhibiting IL-13-induced CD23 expression in a cell, the method comprising contacting the cell with an antibody described herein.

[0454] In certain embodiments, described herein is a method of inhibiting the secretion of IL-13-induced CCL2 and CCL26 from cells, the method comprising contacting the cells with an antibody described herein.

[0455] In certain embodiments, described herein is a method of inhibiting IL-13-induced NTRK1 expression in cells, the method comprising contacting the cells with an antibody described herein.

[0456] In certain embodiments, described herein is a method of reducing the level of thymus and activation-regulated chemokine (TARC) / CCL17 in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0457] In certain embodiments, described herein is a method of preventing an inflammatory disorder or disease in a mammalian subject in need thereof, the method comprising administering to the mammalian subject a therapeutically effective amount of an antibody or pharmaceutical composition described herein.

[0458] Route of Administration In some embodiments, the methods provided herein are useful for treating a disease or disorder in an individual. In one embodiment, the individual is a human and the antibody is an anti-IL-13 antibody described herein.

[0459] In some embodiments, the antibody is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. An effective amount of the anti-IL-13 antibody can be administered for the treatment of a disease or disorder. The appropriate dosage of the anti-IL-13 antibody can be determined based on the type of disease or disorder being treated, the type of anti-IL-13 antibody, the severity and course of the disease or disorder, the condition of the individual, the individual's medical history and response to treatment, and the discretion of the attending physician.

[0460] In some embodiments, the antibodies provided herein are administered with at least one additional therapeutic agent. Any suitable additional therapeutic agent or immunotherapeutic agent can be administered with the antibodies provided herein. Additional therapeutic agents include agents used to treat or prevent a disease or disorder, such as, but not limited to, an inflammatory disease or disorder associated with elevated levels of IL-13 and / or IgE.

[0461] The additional therapeutic agent can be administered by any suitable means. In some embodiments, the antibodies provided herein and the additional therapeutic agent are included in the same pharmaceutical composition. In some embodiments, the antibodies provided herein and the additional therapeutic agent are included in different pharmaceutical compositions.

[0462] In embodiments where the antibodies provided herein and the additional therapeutic agent are included in different pharmaceutical compositions, administration of the antibody can occur before, at the same time as, and / or after administration of the additional therapeutic agent. In some embodiments, administration of the antibodies provided herein and the additional therapeutic agent occurs at intervals within about 1 month. In some embodiments, administration of the antibodies provided herein and the additional therapeutic agent occurs at intervals within about 1 week. In some embodiments, administration of the antibodies provided herein and the additional therapeutic agent occurs at intervals within about 1 day. In some embodiments, administration of the antibodies provided herein and the additional therapeutic agent occurs at intervals within about 2 hours. In some embodiments, administration of the antibodies provided herein and the additional therapeutic agent occurs at intervals within about 1 hour.

[0463] Kits and Products This application provides a kit comprising any one or more of the antibody compositions described herein and an instruction manual. In some embodiments, the kit further comprises a component selected from any of a secondary antibody, immunohistochemical analysis reagents, pharmaceutically acceptable excipients, and handling instructions, and any combination thereof. In one particular embodiment, the kit comprises a pharmaceutical composition comprising any one or more of the antibody compositions described herein together with one or more pharmaceutically acceptable excipients.

[0464] This application also provides a product comprising any one of the antibody compositions or kits described herein. Examples of products include vials (including sealed vials).

Examples

[0465] The following are examples of specific embodiments for practicing the present invention. These examples are presented for illustrative purposes only and are not intended to limit the scope of the present invention in any way. Regarding the numerical values used (e.g., amounts, temperatures, etc.), efforts have been made to ensure accuracy, but of course, some experimental errors and deviations should be taken into account.

[0466] The practice of the present invention uses conventional methods of protein chemistry, biochemistry, recombinant DNA technology, and pharmacology within the skill of those in the art unless otherwise indicated. Such techniques are well described in the literature. For example, T.E. Creighton, Proteins: Structures and Molecular Properties (W.H. Freeman and Company, 1993), A.L. Lehninger, Biochemistry (Worth Publishers, Inc., current addition), Sambrook, et al., Molecular Cloning: A Laboratory Manual (2 ndEdition, 1989), Methods In Enzymology (S. Colowick and N. Kaplan eds., Academic Press, Inc.), Remington’s Pharmaceutical Sciences, 18 th Edition (Easton, Pennsylvania: Mack Publishing Company, 1990), Carey and Sundberg Advanced Organic Chemistry 3 rd Ed. (Plenum Press) Vols A and B (1992). See also ,

[0467] , .

[0467] Method Humanization of the Mouse Hybridoma Sequence of Anti-IL-13 Antibody 228B / C-1 Using complementarity-determining region (CDR) grafting technology, the parental mouse anti-human IL-13 228B / C-1, the parental monoclonal antibody of lebrikizumab, was humanized. The heavy and light chain sequences of the parental mouse were modeled on human antibody frameworks as described below. A set of human heavy and light chains was selected for humanization. The goal was to design pairs of these heavy and light chains that would improve the biophysical properties of the parental antibody while retaining binding. These humanized molecules were designed to improve the developability profile during the scale-up process in bioprocesses.

[0468] Humanization of the Light Chain The parental mAb light chain sequence of the mouse hybridoma sequence of anti-IL13 antibody (lebrikizumab) was compared with a group of human variable region light chain (VK) germline amino acid sequences (Lefranc, M.-P. IMGT, the international ImMunoGeneTics database Nucleic Acids Res., 29, D207-209 (2001). DOI: 10.1093 / nar / 29.1.207. PMID: 11125093.). A total of four human VK germlines were selected. Of these, one belonged to the Vk4 family (IGKV4-1), two belonged to the VK1 family (IGKV1-39 and IGKV3-15), and one belonged to the VK2 family (IGKV2-28). In light chain framework 3, one substitution from R to G was also designed. This human-to-mouse back mutation could alter the binding. The human germline KJ4 was selected as the J region based on sequence similarity to the mouse sequence. The humanized VL domain was cloned into a vector encoding the kappa light chain constant domain.

[0469] The following nomenclature was used for the light chain: "LC0" corresponds to the mouse hybridoma sequence. "LC1" corresponds to IGKV4-1_KJ4. "LC2" corresponds to IGKV1-39_KJ4. "LC3" corresponds to IGKV3-15_KJ4. "LC4" corresponds to IGKV2-28_KJ4. "LC5" corresponds to IGKV4-1_R~G_KJ. "LC6" corresponds to IGK V1-39_R~G_KJ4. "LC7" corresponds to IGKV3-15_R~G_KJ4. "LC8" corresponds to IGKV2-28_R~G_KJ4.

[0470] Humanization of the heavy chain The parental mAb heavy chain sequence of the mouse hybridoma sequence of anti-IL13 antibody (lebrikizumab) was compared with a group of human variable region heavy chain (VH) germline amino acid sequences. A total of five human VH germlines were selected. Of these, one belonged to the VH4 family (IGHV4-59), two belonged to the VH1 family (IGHV1-46, IGHV1-69), and two belonged to the VH3 family (IGHV3-15, IGHV3-23). The Q at the N-terminus of the heavy chain was replaced with E to avoid the possibility of pyroglutamic acid conversion. The human germline HJ6 was selected as the J region based on sequence similarity to the mouse sequence. The humanized VH domain was cloned into a vector encoding the human IgG1 HC constant domain.

[0471] The following nomenclature was used for the heavy chains: "HC0" corresponds to the mouse hybridoma heavy chain. "HC0_M" corresponds to HC0_NIS~TIS of FR3 (to avoid the possibility of glycosylation). "HC1" corresponds to the humanized sequence IGHV4-59_HJ6. "HC2" corresponds to the humanized sequence IGHV1-46_HJ6. "HC3" corresponds to the humanized sequence IGHV1-69_HJ6. "HC4" corresponds to the humanized sequence IGHV3-15_HJ6. "HC5" corresponds to the humanized sequence IGHV3-23_HJ6.

[0472] Gene synthesis and plasmid construction The coding sequences of the antibody HC and LC were generated by DNA synthesis and PCR and then subcloned into pTT5-based plasmids for protein expression in mammalian cell lines. The gene sequences of these expression vectors were confirmed by DNA sequencing.

[0473] Expression of antibody constructs Transient expression of the antibody was performed by co-transfecting pairs of HC and LC constructs into CHO cells using the PEI method. Briefly, in a shake flask, approximately 5.5×10 6CHO cells at [X] cells / mL were used as the host. Transfection was initiated by adding OptiMEM™ medium (Invitrogen) containing a mixture of 1 mg / L DNA and 7 mg / L PEI to the cells and then gently mixing. The cells were then cultured in an incubator shaker at 120 rpm, 37 °C, and 8% CO2 for 9 days. The supply of peptone and glucose was carried out 24 hours later and then every 2 - 3 days depending on cell density and viability. Cell culture was terminated on day 9 when the cell viability decreased below 80%. This conditioned medium was collected for protein purification.

[0474] Purification of the antibody construct Protein purification by affinity chromatography and ion exchange chromatography was performed using an AKTA pure instrument (GE Lifesciences). The conditioned medium expressing the target antibody was harvested by centrifugation at 4000 rpm for 50 minutes and filtered through a 0.22 μm filter. The harvested supernatant was loaded onto a column of Mabselect™ SuRe™ (GE Healthcare). 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 was then buffer-exchanged into 20 mM sodium acetate pH 5.5. [Table 11 - 1] [Table 11 - 2]

[0475] SEC - HPLC analysis of the antibody construct Analytical SEC-HPLC was performed using a Shimadzu LC-10 HPLC apparatus (Shimadzu Corp.). 20 μl of a 1 mg / mL sample was loaded onto a Superdex® 200 Increase 5 / 150GL column (GE Lifesciences). The mobile phase was 2*PBS, flow rate 0.3 mL / min, for 15 minutes.

[0476] Measurement of the antibody-IL13 binding reaction rate using surface plasmon resonance Using a Biacore 8K SPR system (GE HealthCare) equipped with a Series S sensor chip Protein G (Cytiva, catalog 29179315), the binding reaction rate and affinity constant were measured at 25 °C and in a running buffer of HBS-EP+ (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% Surfactant P20). Following the stabilization time in the running buffer, the anti-IL13 mAb construct (diluted to 1 μg / mL) was captured onto Flow Cell 2 (active) at a flow rate of 10 μL / min for 60 seconds. Recombinant human IL-13 protein, His Tag (Acro catalog 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 into Flow Cell 1 (reference) and Flow Cell 2 (active) at a flow rate of 30 μL / min for 180 seconds. Recombinant cynomolgus IL-13 protein, His Tag (SINO BIOLOGICAL, catalog 11057-C07H) was prepared at concentrations of 0, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25, and 0 nM and injected into Flow Cell 1 (reference) and Flow Cell 2 (active) at a flow rate of 30 μL / min for 180 seconds. Samples were injected in multiple cycles onto the newly captured mAb by injecting glycine pH 1.5 at a flow rate of 30 μL / min for 30 seconds to regenerate the capture surface. The data were processed and analyzed with Biacore Insight Evaluation Software version 2.0.15.12933 (GE Healthca) as follows. The response from Flow Cell 1 (reference) was subtracted from the response from Flow Cell 2 (active). Next, the response from two blank injections of buffer was subtracted from the reference subtracted data (2 - 1) to obtain double reference data, which was fit to a 1:1 binding model to identify the apparent association rate constant (ka) and dissociation rate constant (kd). Their ratio provided the apparent equilibrium dissociation constant or affinity constant (KD = kd / ka).

[0477] Measurement of Antibody Affinity for Fc Receptor and C1q The binding affinity (K D ) for Fc receptors and C1q was measured by surface plasmon resonance (SPR) using a Biacore 8K. Briefly, a purified antibody normalized to 5 mg / mL was captured for 90 seconds or 120 seconds at a flow rate of 10 μL / min using an SPR chip functionalized with an anti-kappa light chain antibody. A buffer-only reference channel was used as a reference. Subsequently, various concentrations of recombinant human CD32a (167H), CD32a (167R), CD32b, CD16a (176V), CD16a (176F), FcRn, CD64, and C1q were injected into the surface with the captured purified antibody and the reference channel. Chip regeneration between different concentrations of different antigens was performed using 10 mM glycine HCl, pH 1.5, and the antibody was captured again. The association rate constant and dissociation rate constant were then determined by fitting to either a 1:1 Langmuir binding model or a steady-state analysis model, and the K D value was derived using the Biacore Insight Evaluation Software.

[0478] Evaluation of blockade by cell line-based assay To evaluate the blockade of the complete signal transduction complex of IL-13 / IL13Rα1 / IL-4Rα and the inhibition of downstream signal transduction, multiple assays were used. Briefly, HEK293 cells that had been pre-transduced to stably express both hIL-13Rα and hIL-4Rα were cultured and harvested. The cells were seeded at 200,000 cells per well in 100 μL. The cells were washed and the supernatant was discarded. A 100 μL mixture (volume ratio 1:1) of biotinylated hIL-13 and purified antibody that had been prepared in advance and incubated for 1 hour was added to resuspend the cells, resulting in a final concentration of 0.05 μg / mL of hIL-13 and 0 - 100 nM of purified antibody. The cells were stained in this mixture at 4°C for 1 hour. Subsequently, the cells were washed and stained with 100 μL of Alexa Fluor 488-conjugated streptavidin diluted 1:1000 to detect the binding of biotinylated hIL-13 on the cell surface. The cells were incubated at 4°C for 1 hour in the dark. Then the cells were washed, and the median fluorescence intensity (MFI) of the cells in each well was recorded by FACS using a BD FACSCanto II. The subsequent data was analyzed using GraphPad Prism. The IC 50 value was defined as the concentration of antibody required to inhibit 50% of the maximum MFI of the biotinylated hIL-13 surface detected in the incubation with 0.05 μg / mL of hIL-13 alone.

[0479] Cell line-based assays included the inhibition of STAT6 phosphorylation in HT-29 cells, the inhibition of TARC release in A549 cells, and the inhibition of TF-1 cell proliferation. Primary human lymphocyte-based assays included the inhibition of STAT6 phosphorylation and the inhibition of CD23 expression.

[0480] Inhibition of STAT6 phosphorylation in HT-29 cells The functional activity of an antibody that blocks IL-13-induced bioactivity was evaluated using inhibition of STAT6 phosphorylation in HT-29 cells. Briefly, HT-29 cells were starved overnight in RPMI 1640 + 0.1% FBS. Cells were harvested and seeded at 50,000 cells per 100 μL per well. Simultaneously, 100 μL of a mixture of hIL-13 and purified antibody (1:1 by volume) was added to the same wells to obtain a final concentration of 10 ng / mL of hIL-13 and 0 - 50 nM of purified antibody. Cells were incubated at 37 °C for 1 hour, then fixed, permeabilized, and stained with a PE-conjugated anti-pSTAT6 antibody. The MFI of the cells in each well was recorded by FACS using a BD FACSCanto II, and the subsequent data were analyzed using GraphPad Prism. The IC50 value was determined as the concentration of antibody required to inhibit 50% of the maximum MFI of pSTAT6 detected in the incubation with 10 ng / mL of hIL-13 alone. The results are summarized below.

[0481] Inhibition of TARC release in A549 cells The functional activity of an antibody that blocks IL-13-induced bioactivity was evaluated using inhibition of TARC secretion by A549 cells. Briefly, A549 cells were seeded at 20,000 cells in 100 μL of DMEM + 10% FBS and cultured overnight at 37 °C. The next day, the medium of the cells was discarded and the cells were gently washed with fresh medium. 150 μL of a mixture of hIL-13, purified antibody, and hTNFα (1:1:1 by volume) was added to the wells to obtain a final concentration of 20 ng / mL of hIL-13, 0 - 100 nM of purified antibody, and 200 ng / mL. Cells were incubated in this mixture at 37 °C for 20 - 24 hours. After incubation, the culture supernatant was collected and the amount of TARC present was analyzed using a commercially available TARC ELISA kit (R&D Systems) and analyzed according to the manufacturer's instructions. The TARC concentration in each well measured was analyzed using GraphPad Prism. The IC50 value was determined as the concentration of antibody required to inhibit 50% of the maximum TARC concentration detected in the incubation with 20 ng / mL of hIL-13 and 200 ng / mL of hTNFα alone. The results are summarized below.

[0482] Inhibition of the proliferation of TF-1 cells Using the proliferation of TF-1 cells or its inhibition, the functional activity of an antibody that blocks IL-13-induced bioactivity was evaluated. Briefly, TF-1 cells were harvested and starved for 4 hours in RPMI 1640 + 10% FBS without additional cytokines. During this time, a mixture of hIL-13 and the purified antibody (volume ratio 1:1) was prepared and 50 μL per well was added. After starvation, TF-1 cells were harvested again and seeded at 15,000 cells per 50 μL well to obtain a final concentration of 4 ng / mL of hIL-13 and 0 - 5 nM of the purified antibody. Then, the cells were incubated at 37 °C for 72 hours, and cell proliferation was quantified using CellTiter-Glo (Promega) according to the manufacturer's instructions. Luminescence was recorded with a SpectraMax M5 multimode plate reader, and the data were analyzed using GraphPad Prism. The IC50 value was identified as the concentration of the antibody required to result in 50% of the maximum luminescence detected when TF-1 cells were incubated and cultured with 4 ng / mL of hIL-13 alone. The results are summarized below.

[0483] Inhibition of STAT6 phosphorylation and CD23 expression in primary human lymphocytes To confirm the antagonist activity of the antibody herein in primary cells, human peripheral blood mononuclear cells (PBMCs) obtained from healthy donors were used to evaluate the ability of the antibody herein to inhibit the phosphorylation of IL-13-induced STAT6 and the upregulation of CD23 expression.

[0484] Frozen human PBMCs from a pre-identified IL-13-responsive donor were thawed and allowed to recover. To measure the in vitro potency of the antibodies herein when inhibiting STAT6 phosphorylation, total PBMCs were stimulated with 10 ng / mL of IL-13 and purified antibody (volume ratio 1:1). These cells were incubated at 37 °C for 15 minutes. Thereafter, phosphorylation of STAT6 was analyzed by flow cytometry using a commercially available anti-phospho STAT6 antibody, and staining was evaluated within specific immune populations gated by lineage-specific markers CD14 and CD19. In another set of PBMCs, the cells were incubated at 37 °C for 24 hours. Thereafter, CD23 expression was analyzed by flow cytometry using an anti-CD23 antibody, and the intensity of that staining was evaluated within specific immune populations gated by lineage-specific markers CD14 and CD19.

[0485] Data on the median fluorescence intensity (MFI) from either pSTAT6 or CD23 staining were analyzed using GraphPad Prism. IC 50 values were determined as the concentration of antibody required to bring about 50% of the maximum MFI detected for each marker when primary human PBMCs were incubated and cultured with 10 ng / mL of hIL-13 alone.

[0486] Protein thermal stability testing by differential scanning fluorimetry (DSF) SYPRO® Orange (Thermo Fisher #S6651) was supplied at a 5000-fold concentration in 100% DMSO and diluted 40-fold in an appropriate formulation buffer. The antibody was mixed with this dye, and 9 microliters of this mixture was loaded into a UNi (Unchained Labs, catalog number 201-1010), and run on the UNCLE (Unchained Labs) with the "T m using SYPRO" application. Samples were subjected to a temperature ramp from 25 to 95 °C at a lamp rate of 0.5 °C / min and excitation at 473 nm. The entire spectrum was collected from 250 to 720 nm, and the area under the curve from 510 to 680 nm was measured using UNCLE software to determine the inflection point (Tm ) was calculated.

[0487] HIC-HPLC analysis of the antibody construct HIC-HPLC analysis was performed using a Thermo UltiMate™ 3000 instrument. 20 μl of a 1 mg / mL sample was loaded onto a Thermo Scientific™ MAbPac™ HIC-Butyl HPLC column (5 μm, 4.6 mm × 100 mm, catalog number 088558). Mobile phase A was 1.5 M ammonium sulfate + 50 mM PB buffer + 5% (v / v) isopropyl alcohol, pH 6.95, and mobile phase B was 50 mM PB buffer + 20% (v / v) isopropyl alcohol, pH 6.95. The gradient was 0% - 100% mobile phase B over 20 minutes, and the flow rate was 0.5 mL / min. Inhibition of CCL26 and CCL2 secretion and NTRK1 expression

[0488] The functional activity of an antibody that blocks IL-13-induced bioactivity was evaluated using inhibition of CCL26 and CCL2 secretion and NTRK1 expression by HaCaT cells. 20,000 HaCaT cells were seeded in 100 μL of DMEM + 10% FBS and cultured overnight at 37°C. The next day, 150 μL of a mixture of hIL-13 and the purified antibody was added to the wells to obtain a final concentration of 50 ng / mL of IL-13 and 0 - 206.5 nM of the purified antibody. Then, the cells were incubated for an additional 48 hours at 37°C. After incubation, the culture supernatant was collected, and the levels of secreted CCL26 and CCL2 were measured using a commercially available Luminex-based immunoassay kit (R&D Systems) and analyzed according to the manufacturer's instructions. The concentrations of CCL26 and CCL2 in each well measured were analyzed using GraphPad Prism. The IC 50 value was determined as the concentration of antibody required to inhibit 50% of the maximum concentration detected in the incubation with 50 ng / mL of IL-13 alone.

[0489] The cells remaining on the assay plate were lysed, and mRNA was extracted to analyze NTRK1 gene expression using a commercially available Quantigene kit (ThermoFisher). The level of NTRK1 mRNA was determined according to the manufacturer's protocol and analyzed with GraphPad Prism. NTRK1 gene expression was quantified as the ratio of the NTRK1 mRNA level to the housekeeping gene PPIB, and the IC 50 value was calculated as the concentration of antibody required to inhibit 50% of the maximum gene expression detected using 50 ng / mL of hIL-13 alone.

[0490] Example 1: Engineered anti-IL-13 antibodies exhibit improved affinity and blocking potency for IL-13 Results Measurement of antibody affinity for IL-13 Using the method described above, the affinity of construct 133 (see construct sequences in Tables 2-8) for IL-13 and its binding reaction rate were evaluated using surface plasmon resonance (SPR) and compared to dupilumab, lebrikizumab, and a variant of lebrikizumab having one or more amino acid substitutions in the heavy chain constant region (construct 2 (Lebrikizumab-HC, Lebrikizumab-LC, hIgG1-LAGA YTE, human kappa LC), see construct sequences in Tables 2-8), as well as tralokinumab.

[0491] As measured by SPR, the affinity of construct 133 was 77 pM, compared to 131 pM and 116 pM for lebrikizumab and tralokinumab, respectively.

[0492] The affinity of variants of lebrikizumab having one or more amino acid substitutions in the heavy chain constant region (constructs 128 - 131), variants of construct 15 or 98 having one or more amino acid substitutions in the heavy chain constant region (constructs 133 - 136 or 137 - 140, respectively), and variants having one or more amino acid substitutions in the heavy chain constant region (constructs 132 and 141 - 144) for human and cynomolgus monkey IL-13, as well as their binding reaction rates, were also evaluated using SPR.

[0493] All antibodies were observed to bind human IL-13 with low picomolar affinity comparable to variants of lebrikizumab. Furthermore, all antibodies tested were cross-reactive with cynomolgus monkey IL-13 with sub-nanomolar affinity (Table 12). [Table 12]

[0494] Antibody Affinity for Fc Receptors and C1q Compared to lebrikizumab, all antibody hIgG1-LALA YTE variants had binding to all Fc gamma receptors nearly or completely eliminated, binding to C1q significantly decreased, and binding to FcRn significantly increased at pH 5.8 (Table 13). [Table 13]

[0495] Blocking A. Inhibition of IL-13 Binding to hIL-13Rα / hIL-4Rα Overexpressing Cells The functional blockade of antibodies against this binding interaction was evaluated using IL-13 binding to cells overexpressing hIL-13Rα / hIL-4Rα. The results of the functional blockade of the antibodies described herein that block IL-13 binding to cells overexpressing hIL-13Rα / hIL-4Rα are shown in Table 14 and Figure 2. In the cell line-based assay, the IC of 1.11 nM for lebrikizumab 50Compared with [comparison object], construct 133 showed an IC of 0.89 nM and inhibited IL-13 binding in the IL-13Rα1 / IL-4Rα overexpressing cell line. 50 and inhibited IL-13 binding in the IL-13Rα1 / IL-4Rα overexpressing cell line. [Table 14]

[0496] B. Inhibition of IL-13-induced STAT6 phosphorylation in HT-29 cells The functional activity of antibodies that block IL-13-induced bioactivity was evaluated using the inhibition of STAT6 phosphorylation in HT-29 cells. Regarding the inhibition of STAT6 phosphorylation in HT-29 cells, compared with the observed IC of 0.28 nM for construct 133, it was 0.16 nM for dupilumab, 0.23 nM for lebrikizumab, and 0.41 nM for tralokinumab, respectively. 50 and 0.23 nM for lebrikizumab, and 0.41 nM for tralokinumab, respectively, compared with the observed IC of 0.28 nM for construct 133.

[0497] Variants of lebrikizumab with one or more amino acid substitutions in the heavy chain constant region (constructs 128 - 131), variants of construct 15 or 98 with one or more amino acid substitutions in the heavy chain constant region (constructs 133 - 136 or 137 - 140, respectively), and variants with one or more amino acid substitutions in the heavy chain constant region (constructs 132 and 141 - 144) were also examined in the same assay (Table 15 and Figure 3). [Table 15]

[0498] C. Inhibition of TARC secretion of IL-13 by engineered anti-IL13 antibody variants Human A549 cells express the IL4Rα / IL13Rα1 receptor, which responds to binding of human IL-13 by inducing phosphorylation of pSTAT6, thereby inducing the expression of downstream genes involved in the TH2-type allergic response. Thymus and activation-regulated chemokine (TARC), also known as CCL17, is one of the gene products secreted by multiple cell types and plays a role in attracting effector immune cells such as eosinophils involved in inflammation. A549 cells were contacted with engineered anti-IL-13 antibodies and a TARC assay was performed (Figure 4). The anti-IL-13 antibodies inhibited the secretion of TARC as measured by ELISA. The IC 50 profile (Table 16) was similar to that of lebrikizumab, and thus it was confirmed that the potency of the anti-IL13 antibodies against IL-13 capture activity was maintained in the cell-based assay.

Table 16

[0499] Variants of lebrikizumab having one or more amino acid substitutions in the heavy chain constant region (Constructs 128 - 131), variants of Construct 15 or 98 having one or more amino acid substitutions in the heavy chain constant region (Constructs 133 - 136 or 137 - 140, respectively), and variants having one or more amino acid substitutions in the heavy chain constant region (Constructs 132 and 141 - 144) were examined in the same assay (Table 17 and Figure 5). For the inhibition of TARC release in A549 cells, the IC 50 of 0.86 nM for Construct 133 was compared with 1.11 nM for dupilumab, 0.74 nM for lebrikizumab, and 4.14 nM for tralokinumab, respectively.

Table 17

[0500] D. Inhibition of the proliferation of IL-13-induced TF-1 cells Regarding the inhibition of the proliferation of IL-13-induced TF-1 cells, the IC of 0.16 nM by construct 133 50 was observed. In comparison, it was 0.19 nM for dupilumab, 0.20 nM for lebrikizumab, and 0.59 nM for tralokinumab, respectively.

[0501] Variants of lebrikizumab with one or more amino acid substitutions in the heavy chain constant region (constructs 128 - 131), variants of construct 15 or 98 with one or more amino acid substitutions in the heavy chain constant region (constructs 133 - 136 or 137 - 140, respectively), and variants with one or more amino acid substitutions in the heavy chain constant region (constructs 132 and 141 - 144) were also examined in the same assay (Table 18 and Figure 6). [Table 18]

[0502] E. Inhibition of IL-13-induced STAT6 phosphorylation and CD23 expression in primary human lymphocytes In primary human lymphocytes, construct 133 potently blocked IL-13 activity in a dose-dependent manner, with an IC of 0.44 nM in the inhibition of STAT6 phosphorylation 50 compared to 0.38 nM for lebrikizumab, and an IC of 0.85 nM in the inhibition of CD23 expression 50 compared to 0.81 nM for lebrikizumab. These results demonstrated the potent antagonist activity of construct 133 against IL-13-mediated signaling in primary human cells.

[0503] Variants of construct 98 with one or more amino acid substitutions in the heavy chain constant region (construct 137), and variants with one or more amino acid substitutions in the heavy chain constant region (construct 141) were also examined in the same assay (Table 19, Figure 7, and Figure 8). [Table 19]

[0504] Example 2: Expression of the engineered anti-IL-13 antibody was higher than that of lebrikizumab Transient expression of the antibody was performed by co-transfecting CHO cells with pairs of HC and LC constructs using the PEI method described above. Next, the relative expression of the engineered antibody constructs from CHO cell lysates compared to lebrikizumab was determined in a small-scale expression screening experiment (Table 20). [Table 20-1] [Table 20-2]

[0505] Example 3: Thermal stability of the engineered anti-IL-13 antibody Differential scanning fluorimetry (DSF) was used to measure the melting temperature (Tm2) of lebrikizumab and the IL-13 antibodies described in Table 21. Most of the variants showed a Tm2 profile comparable to that of lebrikizumab, but a few variants, such as Construct 5 and Construct 15, showed significantly higher melting temperatures (Table 21). Furthermore, almost all variants showed a higher aggregation temperature Tagg compared to lebrikizumab. Tagg is a measure of the tendency of the mAb to form high molecular weight aggregates, and higher values are more desirable. Thus, most of the engineered variants had a more desirable aggregation temperature. [Table 21]

[0506] Example 4: Engineered anti-IL-13 antibodies show reduced hydrophobicity Hydrophobic interaction chromatography (HIC) was performed to measure the tendency of the engineered anti-IL-13 antibodies to interact with hydrophobic surfaces (Table 22). Shorter retention times indicate lower degrees of hydrophobicity. All of the novel IL-13 antibodies examined showed shorter retention times (RT) compared to lebrikizumab. Thus, all of the engineered anti-13 antibodies examined showed a decrease in hydrophobicity compared to lebrikizumab.

Table 22

[0507] Example 6: The engineered anti-IL13 antibody variant and lebrikizumab have the same epitope on IL-13 Epitope binning is a technique that characterizes whether two antibodies specific for the same target (in this case, IL-13) can each bind to the target simultaneously. If an mAb pair blocks each other's ability to bind to the target antigen, they are binned together. An mAb pair that is binned together typically binds to the same or overlapping epitopes on the antigen.

[0508] To characterize the binding of construct 133 containing SEQ ID NOs: 3, 39, 439, and 469 in comparison to lebrikizumab, lebrikizumab was immobilized on a sensor chip surface capable of measuring mAb-antigen interactions. First, when IL-13 was injected into the flow channel, a response was generated due to the binding of IL-13 to lebrikizumab. Subsequently, when construct 133 was injected into the flow channel, a response of the interaction was recorded. In these tests, no response was observed after the injection of construct 133 (see the results in Table 23). This indicates that construct 133 and lebrikizumab are binned together, providing evidence supporting the high likelihood that these two mAbs bind to similar or the same epitope of IL-13.

Table 23

[0509] In a similar test, tralokinumab-ldrm (Adbry™) was found to have a non-binning response, suggesting that it has a different epitope on IL-13 from lebrikizumab.

[0510] To further characterize the constructs 133 and the epitopes of lebrikizumab, hydrogen-deuterium exchange mass spectrometry (HDX-MS) and cross-linking mass spectrometry (XL-MS), which are known in the art, were performed. Briefly, HDX-MS was performed using IL-13 alone at a concentration of 20 μM or a mixed solution of IL-13 and a purified antibody at final concentrations of 20 μM and 40 μM, respectively. After incubations of 15 seconds, 60 seconds, 180 seconds, 600 seconds, 1800 seconds, and 7200 seconds, the exchange reaction was quenched and the protein mixture was subjected to LC-MS using Waters Q-ToF Xevo G2-XS following proteolysis. Deuterium incorporation was measured for both IL-13 and the mixture, and peptide regions where deuterium incorporation was inhibited were considered likely to be involved in binding to the purified antibody. Briefly, XL-MS was performed using a mixed solution of IL-13 and the purified antibody at final concentrations of 14.7 μM and 0.7 μM, respectively. 20 μL of this mixture was mixed with 2 μL of DSS (a stock solution of DMF containing 2 mg / mL), and the final solution was incubated at room temperature for 180 minutes. After this incubation, the sample was subjected to proteolysis using trypsin, chymotrypsin, ASP-N, elastase, and thermolysin and analyzed using LC-MS with Q-Exactive MS. Peptides were compared to peptides identified from a prior peptide mapping experiment of IL-13 alone. Peptide regions of IL-13 that showed cross-linking to the corresponding peptides from the antibody variable regions were considered likely to be involved in binding to the purified antibody. Both methods provided complementary data indicating specific amino acid regions to which constructs 133 and lebrikizumab are likely to bind. This provided further evidence supporting the likelihood that the two mAbs bind to highly overlapping epitopes of IL-13 (Figure 1).

[0511] These tests provided evidence that construct 133 binds to the same region of IL-13 as lebrikizumab and, thus, they are likely to have the same biological effects rather than recognizing different regions.

[0512] Example 7: The engineered anti-IL-13 antibody variants demonstrated a significant prolongation of the half-life in NHP and pharmacokinetic analysis of the anti-IL-13 antibody Prolongation of the half-life of anti-IL-13 antibody variants in NHP To demonstrate the potential of engineered anti-IL13 antibody variants to improve dosing compared to the current and planned standard-of-care mAbs in Alzheimer's disease (AD) among other diseases, construct 133 containing SEQ ID NOs: 3, 39, 439, and 469 was examined in female non-human primates (NHP) after a single bolus dose of 3 mg / kg administered either IV or SQ. Blood samples were collected continuously starting from pre-dose samples at 0.167, 1, 4, 8, 24, 48, 96, 168, 336, 504, 674, 840, 1334, 1680, and 2160 hours post-dose. The maximum serum concentration (C max ), time to maximum serum concentration (T max ), area under the serum concentration-time curve extrapolated from time 0 to infinity (AUC 0-inf ), clearance (CL), volume of distribution at steady state (V SS ), half-life (T 1 / 2 ), and absolute subcutaneous bioavailability (F) PK parameters were calculated. The data were analyzed to show the mean serum concentration including the standard deviation over time and a regression fit was performed.

[0513] In a 1:1 study of construct 133 versus lebrikizumab in NHP, both the IV and SQ formulations of construct 133 showed a significantly longer half-life than lebrikizumab. In these studies, as shown in Figure 9, the mean half-life of construct 133 was 27.6 days compared to 18 days for lebrikizumab. Additionally, construct 133 showed an average clearance rate of 1.45 (mL day -1 / kg -1 ) in NHP. The volume of distribution at steady state was found to be 55.65 (mL / kg -1 ). Construct 133 was well-absorbed and the subcutaneous bioavailability was determined to be 81.22%. Lebrikizumab had an average clearance rate of 2.93 (mL day -1 / kg -1) was shown. The steady-state distribution volume was found to be 52.10 (mL / kg -1 ). Lebrikizumab was well absorbed and the subcutaneous bioavailability was determined to be 75.70%. Without being bound by theory, construct 133 was engineered to have an amino acid substitution of YTE in the Fc region, so its IgG half-life may have been extended by increased binding to the neonatal Fc receptor (FcRn) under acidic pH conditions. IgG bound to FcRn is recycled via lysosomal salvage, resulting in IgG returning to circulation. The long half-life of construct 133 may result in a reduced dosing frequency compared to currently available therapies, thereby reducing the injection burden and increasing compliance for patients with atopic dermatitis and other diseases caused by IL-13.

[0514] In a non-one-to-one comparison of NHP data obtained from third parties, construct 133 had the highest normalized AUC among antibodies with a YTE substitution, as shown in Figure 10 0-∞ (C norm*day ), or the area under the curve (AUC) from administration to infinity. Thus, the PK profile of construct 133 appears to provide the highest sustained concentration or drug level in the bloodstream compared to other antibodies with a YTE substitution.

[0515] The extension of the half-life of mAbs with an amino acid substitution of YTE depends on the type of target (e.g., receptor vs. soluble). Thus, for mAbs against soluble targets, when investigating the conversion of NHP half-life data to human half-life data, it was found that the human half-life was approximately 3 to 4 times longer than the NHP half-life (mean 3.5-fold, median 3.1-fold, data not shown).

[0516] Based on this NHP half-life data, the antibodies disclosed herein (e.g., construct 133) are expected to have a human half-life of about 80 to 110 days based on equivalent mAbs with an amino acid substitution of YTE.

[0517] Furthermore, based on PK modeling, at a human half-life of 33 days (which is lower than the lowest half-life of mAbs with YTE amino acid substitutions and soluble targets reported to date, to the knowledge of the present applicant), the antibodies disclosed herein are considered to be effectively administered on a maintenance dosing schedule every two months. At a half-life of 50 days, the antibodies disclosed herein are considered to be effectively administered on a maintenance dosing schedule every three months.

[0518] To understand the maintenance dosing schedules that may be achievable with the antibodies disclosed herein, the known PK parameters of rituximab were used. These PK parameters provided an understanding of how rituximab distributes throughout the body and is eliminated. Based on these known parameters, a two-compartment PK model at first-order absorption, which is the standard for mAbs, was constructed to predict the concentration or drug level over time for both rituximab and the antibodies disclosed herein. The key parameters included clearance (CL) of 0.156 L / day, central volume (Vc) of 4.10 L, absorption rate (ka) of 0.239 day−1, and bioavailability of 85.6%.

[0519] Efficacy in inflammatory conditions, such as in AD, is thought to be determined by C trough , or the minimum concentration of the mAb. Thus, based on the model above, the target C trough of the antibodies disclosed herein was set to be equal to the C trough of rituximab in maintenance with monthly dosing, which was 31.3 mg / L. Considering the overlapping epitopes between rituximab and a particular antibody disclosed herein, and the similarity in potency across multiple in vitro assays, the exposure required for the potential clinical activity of the antibodies disclosed herein can be predicted. To maintain the concentration of the antibodies of the present disclosure above 31.3 mg / L, K eliminationThat is, by modeling the elimination rate constant or the fraction of drug eliminated over a given time, and the half-life, assuming a dose of 300 mg, a half-life of at least 33 days is required to administer the antibody of the present disclosure every two months for maintenance, and a half-life of at least 50 days is required to administer the antibody of the present disclosure every three months for maintenance.

[0520] Therefore, with a human half-life of 33 days, the antibodies disclosed herein are considered to be effectively administered on a maintenance dosing schedule every two months. With a half-life of 50 days, the antibodies disclosed herein are considered to be effectively administered on a maintenance dosing schedule every three months.

[0521] Pharmacokinetic analysis of anti-IL-13 antibodies In further experiments, multiple in vivo pharmacokinetic (PK) studies were conducted to examine lebrikizumab, constructs 15 or 98 with one or more amino acid substitutions in the heavy chain constant region and further variants thereof (constructs 133 - 137 or 137 and 140, respectively), and / or further variants with one or more amino acid substitutions in the heavy chain constant region (constructs 141 and 144).

[0522] The studies were performed using cynomolgus monkeys (Macaca fascicularis). Any matching or subcutaneous (SQ) / intravenous (IV) cohort consisted entirely of either female monkeys weighing 1.5 kg to 2.0 kg or male monkeys weighing 2.9 kg to 3.3 kg. On day 0, the test agent was administered to the animals at a dose of 3 mg / kg for each antibody by IV bolus and / or SQ injection, and serum samples were collected periodically throughout the study.

[0523] The PK parameters were measured from cynomolgus monkey serum samples up to day 56 (1334 hours), and up to day 90 (2160 hours) in a subset of the cohort. The mean PK curves are shown in Figure 11 for IV administration and Figure 12 for SQ administration. This PK analysis demonstrated that constructs 133, 134, 135, 136, 137, 140, 141, and 144 each had an improved half-life and a reduced serum clearance rate compared to lebrikizumab, as reported in Table 24. When bioavailability (F) could be determined, constructs 133, 134, and 141 were shown to have bioavailability equivalent to lebrikizumab (Table 25).

Table 24-1

Table 24-2

Table 25

[0524] Example 8. Inhibition of IL-13-induced secretion of CCL2 and CCL26 and expression of NTRK1 in HaCaT cells Inhibition of the secretion of CCL26 (eotaxin-3) and CCL2 (MCP-1) by HaCaT cells (Figures 13 and 14, respectively) and inhibition of NTRK1 expression (Figure 15) were used to evaluate the functional activity of antibodies that block IL-13-induced bioactivity. 20,000 HaCaT cells were seeded in 100 uL of DMEM + 10% FBS and cultured overnight at 37 °C. The next day, 150 uL of a mixture of hIL-13 and the purified antibody was added to the wells to obtain a final concentration of 50 ng / mL of IL-13 and 0 - 206.5 nM of the purified antibody. The cells were then incubated for an additional 48 hours at 37 °C. After incubation, the culture supernatant was collected, and the levels of secreted CCL26 and CCL2 were measured using a commercially available Luminex-based immunoassay kit (R&D Systems) and analyzed according to the manufacturer's instructions. The concentrations of CCL26 and CCL2 in each well measured were analyzed using GraphPad Prism. The IC 50 value was identified as the concentration of antibody required to inhibit 50% of the maximum concentration detected in the incubation with 50 ng / mL of IL-13 alone.

[0525] The cells remaining in the assay plate were lysed and mRNA was extracted to analyze NTRK1 gene expression using a commercially available Quantigene kit (ThermoFisher). The level of NTRK1 mRNA was determined according to the manufacturer's protocol and analyzed with GraphPad Prism. NTRK1 gene expression was quantified as the ratio of the NTRK1 mRNA level to the housekeeping gene PPIB, and the IC 50 value was calculated as the concentration of antibody required to inhibit 50% of the maximum gene expression detected using 50 ng / mL of hIL-13 alone. The results are summarized in Table 26.

Table 26

[0526] Example 9. Monomer Purity after Affinity Capture from Stable Pools The monomer purity from one-step affinity capture determines the final yield and unit cost of the antibody under cGMP production. To characterize this, briefly, stable pools of CHO were generated separately for each antibody in a workflow leading to the selection of the master cell bank for cGMP production. The affinity capture step was performed using a Mabselect SuRe column. The novel IgG1 variants (e.g., constructs 132, 133, 136, 137, 140, 141, and 144) maintained a clear solution containing less than 15% aggregates after one-step purification. The novel IgG4 variants (constructs 134, 135, 138, 139, 142, and 143) had a milky appearance, showed some precipitation, and were aggregation-sensitive (>68% aggregation) when eluted with an elution buffer of 50 mM sodium citrate, 150 mM sodium chloride, pH 3.0. These novel IgG4 variants had lower aggregation levels when eluted with either 50 mM acetic acid, pH 2.8, or 100 mM sodium acetate, 800 mM arginine, pH 3.5. At the higher pH 3.5, arginine was required to maintain an appropriate recovery rate.

[0527] The results are summarized in Table 27. The novel IgG1 and IgG4 variants showed high monomer purity directly from affinity capture under optimized elution conditions when compared to the rituximab variants (constructs 128 - 131).

Table 27

[0528] Example 10. Accelerated Stability As demonstrated by this example, the novel variants in the IgG1 construct, as shown in Table 28, had a lower tendency to aggregate and were more resistant to changes in basic species under various stress conditions. The proportion of basic species was determined by capillary isoelectric focusing (ciEF) as is known in the art. Variants related to rituximab (constructs 128 - 131) and IgG4 variants (constructs 134, 135, 142, and 143, as described in Examples 1 - 7) were more prone to aggregation and more susceptible to the effects of changes in basic species compared to the novel IgG1 variants (e.g., constructs 132, 133, 136, 137, 140, 141, and 144).

Table 28

[0529] Although the present invention has been specifically shown and described with reference to preferred embodiments and various alternative embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made without departing from the spirit and scope of the invention.

[0530] All references, issued patents, and patent applications cited in the body of this specification are hereby incorporated by reference in their entirety for all purposes.

Table 29 - 1

Table 29 - 2

Table 29 - 3

Table 29 - 4

Table 29 - 5

Table 29 - 6

Table 29-7

Table 29-8

Table 29-9

Table 29-10

Table 29-11

Table 29-12

Table 29-13

Table 29-14

Table 29-15

Table 29-16

Table 29-17

Table 29-18

Table 29-19

Table 29-20

Table 29-21

Table 29-22

Table 29-23

Table 29-24

Table 29-25

Table 29-26

Table 29-27

Table 29-28

Table 29-29

Table 29-30

Table 29-31

Table 29-32

Table 29-33

Table 29-34

Table 29-35

Table 29-36

Table 29-37

Table 29-38

Table 29-39

Table 29-40

Table 29 - 41

Table 29 - 42

Table 29 - 43

Table 29 - 44

Table 29 - 45

Table 29 - 46

Table 29 - 47

Table 29 - 48

Table 29 - 49

Table 29 - 50

Table 29 - 51

Table 29 - 52

Table 29 - 53

Table 29 - 54

Table 29 - 55

Table 29 - 56

Table 29-57

Claims

1. An isolated antibody that binds to interleukin 13 (IL-13), wherein the antibody comprises a heavy chain variable region sequence described in SEQ ID NO: 3 and a light chain variable region sequence described in SEQ ID NO:

39.

2. The isolated antibody according to claim 1, wherein the antibody comprises a constant heavy chain sequence selected from any one of the sequences described in SEQ ID NOs: 425-468 and 484-539.

3. The isolated antibody according to claim 2, wherein the antibody comprises the constant heavy chain sequence described in SEQ ID NO: 439, SEQ ID NO: 446, SEQ ID NO: 457, SEQ ID NO: 460, or SEQ ID NO:

440.

4. An isolated antibody that binds to IL-13, wherein the antibody comprises a heavy chain variable region sequence described in SEQ ID NO: 3 and a light chain variable region sequence described in SEQ ID NO: 39, and the antibody comprises the Fc region of human IgG1 having a LALA and / or YTE mutation.

5. An isolated antibody that binds to IL-13, wherein the antibody is CDR-H1 containing the sequence described in Sequence ID No. 58, CDR-H2 containing the sequence described in Sequence ID No. 100, CDR-H3 containing the sequence described in Sequence ID No. 112, CDR-L1 containing the sequence described in Sequence ID No. 141, CDR-L2 containing the sequence described in Sequence ID No. 153, and CDR-L3 containing the sequence described in Sequence ID No. 165 Includes, The antibody comprises the Fc region of human IgG1 having LALA and / or YTE mutations. The antibody.

6. The isolated antibody according to claim 4 or 5, wherein the antibody comprises the Fc region of human IgG1 having LALA and YTE mutations.

7. The isolated antibody according to claim 4 or 5, wherein the antibody comprises the Fc region of human IgG1 having a YTE mutation.

8. The isolated antibody according to claim 4 or 5, wherein the antibody comprises the Fc region of human IgG1 having a LALA mutation.

9. An isolated antibody that binds to IL-13, wherein the antibody is CDR-H1 containing the sequence described in Sequence ID No. 58, CDR-H2 containing the sequence described in Sequence ID No. 100, CDR-H3 containing the sequence described in Sequence ID No. 112, CDR-L1 containing the sequence described in Sequence ID No. 141, CDR-L2 containing the sequence described in Sequence ID No. 153, and CDR-L3 containing the sequence described in Sequence ID No. 165 Includes, Image: 、T256E、T256D、T250Q、H285D、T307A、 T307Q、T307R、T307W、L309D、Q411H、Q311V、A378V、E380A、M428L、N434A、N43 4S、N297A、D265A、L234A、L235A、N434 W、S228P(SP)、M428L / N434S(LS)、M25 2Y / S254T / T256E(YTE)、T250Q / M428L、T307A / E380A / N434A、T256D / T307Q(D Q)、T256D / T307W(DW)、M252Y / T256D( YD)、T307Q / Q311V / A378V(QVV)、T256 D / H285D / T307R / Q311V / A378V(DDRVV )、L309D / Q311H / N434S(DHS)、S228P / L 235E(SPLE)、L234A / L235A(LALA)、M428L / N434A(LA)、L235A / G237A(LAGA) 、L234A / L235A / G237A((LALAGA)、L234A / L235A / P329G((LALAPG)、N297A、D265 A / YTE、LALA / YTE、LAGA / YTE、LALAGA / YTE、LALAPG / YTE、N297A / LS、D265A / L S、LALA / LS、LALAGA / LS、LALAPG / LS、N 297A / DHS、D265A / DHS、LALA / DHS、LAGA / DHS、LALAGA / DHS、LALAPG / DHS、S228 P(SP) / YTE、SPLE / YTE、SP / LS、SPLE / L S、SP / DHS、SPLE / DHS、N297A / LA、D265 A / LA、LALA / LA、LAGA / LA、LALAGA / LA、L ALAPG / LA、N297A / N434A、D265A / N434 A、LALA / N434A、LAGA / N434A、LALAGA / N434A、LALAPG / N434A、N297A / N434W、 D265A / N434W、LALA / N434W、LAGA / N434 W、LALAGA / N434W、LALAPG / N434W、N29 7A / DQ、D265A / DQ、LALA / DQ、LAGA / DQ、L ALAGA / DQ、LALAPG / DQ、N297A / DW、D26 5A / DW、LALA / DW、LAGA / DW、LALAGA / DW、The antibody comprising an Fc region containing one or more amino acid substitutions selected from the group consisting of LALAPG / DW, N297A / YD, D265A / YD, LALA / YD, LAGA / YD, LALAGA / YD, LALAPG / YD, T307Q / Q311V / A378V (QVV), N297A / QVV, D265A / QVV, LALA / QVV, LAGA / QVV, LALAGA / QVV, LALAPG / QVV, DDRVV, N297A / DDRVV, D265A / DDRVV, LALA / DDRVV, LAGA / DDRVV, LALAGA / DDRVV, and LALAPG / DDRVV.

10. The isolated antibody according to claim 9, wherein the antibody comprises a human IgG4 Fc region having a YTE substitution, a human IgG4 Fc region having an LS substitution, a human IgG1 Fc region having LALA and LS substitutions, or a human IgG1 Fc region having LAGA and YTE substitutions.

11. An isolated antibody according to any one of claims 4, 5, or 9, wherein the LALA mutation is located at positions 235 and 236 (L235A / L236A) by direct numbering, and the YTE mutation is located at positions 253, 255 and 257 (M253Y / S255T / T257E).

12. The isolated antibody according to claim 5 or 9, wherein the antibody comprises the heavy chain variable region sequence described in SEQ ID NO: 3 and the light chain variable region sequence described in SEQ ID NO:

39.

13. The isolated antibody according to any one of claims 1, 4, 5, and 9, wherein the antibody comprises the constant light chain sequence described in Sequence ID No.

469.

14. The isolated antibody according to any one of claims 1, 4, 5, and 9, wherein the antibody comprises the constant heavy chain sequence of SEQ ID NO: 439 and the constant light chain sequence of SEQ ID NO:

469.

15. An isolated antibody that binds to IL-13, comprising the heavy chain variable region sequence described in SEQ ID NO: 3, the light chain variable region sequence described in SEQ ID NO: 39, the constant heavy chain sequence of SEQ ID NO: 439, and the constant light chain sequence described in SEQ ID NO:

469.

16. The isolated antibody according to any one of claims 1, 4, 5, 9, and 15, wherein the antibody is a humanized, human, or chimeric antibody.

17. The isolated antibody according to claim 16, wherein the antibody is a humanized antibody.

18. The isolated antibody according to any one of claims 1, 9, and 17, wherein the antibody comprises a human heavy chain constant region of class IgG and a subclass selected from IgG1, IgG2, IgG3, and IgG4.

19. The isolated antibody according to claim 18, wherein the antibody comprises the Fc region of human IgG1.

20. The isolated antibody according to any one of claims 1, 4, 5, 9, and 15, wherein the antibody is a monoclonal antibody.

21. The isolated antibody according to any one of claims 1, 4, 5, 9, and 15, wherein the antibody binds to the IL-13 sequence described in SEQ ID NO: 472 or SEQ ID NO:

473.

22. The antibody is measured by surface plasmon resonance (SPR) on the IL-13 sequence described in SEQ ID NO: 472 or SEQ ID NO: 473, and K Ｄ Approximately 1, 2, 3, 4, 5, 6, 7, 8, or 9 x 10 －９ An isolated antibody according to any one of claims 1, 4, 5, 9, and 15, which binds at M or less.

23. The antibody, when measured by SPR, corresponds to the IL-13 sequence described in SEQ ID NO: 472 or SEQ ID NO: 473, and K Ｄ Approximately 1×10 －１０ An isolated antibody according to any one of claims 1, 4, 5, 9, and 15, which binds at M or less.

24. The aforementioned antibody was measured against human IL-13 by SPR, and K Ｄ Approximately 1×10 －９ An isolated antibody according to any one of claims 1, 4, 5, 9, and 15, which binds at M or less.

25. The isolated antibody according to any one of claims 1, 4, 5, 9, and 15, characterized in that the antibody is formulated for subcutaneous injection.

26. The isolated antibody according to any one of claims 1, 4, 5, 9, and 15, characterized in that the antibody is formulated for intravenous injection.

27. A composition comprising an isolated antibody according to any one of claims 1, 4, 5, 9, and 15 and a pharmaceutically acceptable excipient.

28. A composition comprising an isolated antibody according to any one of claims 1, 4, 5, 9, and 15 for treating an inflammatory disorder or disease in a mammalian subject requiring treatment of said inflammatory disorder or disease, or a composition comprising an isolated antibody according to any one of claims 1, 4, 5, 9, and 15 and a pharmaceutically acceptable excipient.

29. The composition according to claim 28, wherein the inflammatory disorder or disease is atopic dermatitis.

30. The composition according to claim 29, characterized in that the treatment reduces the severity of the disease in the subject, and the severity of the disease is assessed by the Atopic Dermatitis Disease Severity Outcome Measure.

31. The composition according to claim 28, wherein the inflammatory disorder or disease is asthma, idiopathic pulmonary fibrosis, alopecia areata, chronic sinusitis with nasal polyps, chronic sinusitis without nasal polyps (CRSsNP), eosinophilic esophagitis (EoE), eosinophilic gastrointestinal disorders or diseases (ENID), Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA), prurigo nodosa (PN), chronic spontaneous urticaria (CSU), chronic pruritus of unknown cause (CPUO), bullous pemphigoid (BP), cold urticaria (ColdU), allergic sinus mycosis (AFRS), allergic bronchopulmonary aspergillosis (ABPA), allergic rhinitis, urticaria, allergic dermatitis, or chronic obstructive pulmonary disease (COPD).

32. The composition according to claim 31, wherein the ENDI is selected from the group consisting of eosinophilic gastritis (EoG), eosinophilic enteritis (EoN), eosinophilic colitis (EoC), and eosinophilic gastroenteritis (EGE).

33. A kit, wherein the kit is (a) an isolated antibody conjugated to IL-13 as described in any one of claims 1, 4, 5, 9, and 15, or a composition comprising an isolated antibody as described in any one of claims 1, 4, 5, 9, and 15 and a pharmaceutically acceptable excipient, and (b) Instructions for use The kit includes the above.

34. An isolated polynucleotide or set of polynucleotides encoding an isolated antibody according to any one of claims 1, 4, 5, 9, and 15.

35. A vector or set of vectors comprising a polynucleotide or set of polynucleotides as described in Claim 34.

36. A host cell comprising a polynucleotide or set of polynucleotides as described in Claim 34, or a vector or set of vectors comprising a polynucleotide or set of polynucleotides as described in Claim 34.

37. The host cell according to claim 36, wherein the host cell is a Chinese hamster ovary (CHO) cell.

38. A method for producing an antibody, the method comprising expressing the antibody using the host cells described in claim 36, and isolating the expressed antibody.

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