Human interleukin-4 receptor alpha antibody

Abstract

The present disclosure relates to antibodies that specifically bind to human IL-4Rα, compositions comprising such IL-4Rα antibodies, and methods of using such IL-4Rα antibodies.

Description

[Technical Field]

[0001] The present disclosure is in the field of medicine. In particular, the present disclosure relates to antibodies that specifically bind to the human interleukin-4 (IL-4) receptor alpha subunit (IL-4Rα), compositions comprising such IL-4Rα antibodies, and methods of using such IL-4Rα antibodies. [Background technology]

[0002] The ongoing epidemic of immunoinflammatory disorders, particularly type 2 inflammatory diseases such as atopic dermatitis, asthma, and food allergies, represents a heterogeneous set of disorders affecting different target tissues. Immunological and genetic studies have identified various cytokines and cytokine receptors involved in type 2 inflammatory diseases, and interleukin-4 receptor α and the related cytokines interleukin-4 and interleukin-13, in particular, have been identified as key drivers of the type 2 inflammatory pathway.

[0003] The human interleukin-4 receptor alpha (IL-4Rα, also known as IL-4 receptor subunit alpha; CD124, BSF receptor) is a transmembrane glycoprotein of the class I cytokine receptor family that plays an important role in diverse biological processes, including Th2-based immune responses, alternative macrophage and dendritic cell activation, mucosal immunity, allergic inflammation, tumor progression, and atherogenesis. IL-4Rα is reported to be expressed on T and B lymphocytes, eosinophils, basophils, monocytes and macrophages, dendritic cells, endothelial cells, fibroblasts, airway epithelial cells, and smooth muscle cells. IL-4Rα exists in two distinct complexes throughout the body: type I receptors consist of an IL-4Rα subunit with a common γ chain and specifically bind IL-4, whereas type II receptors consist of an IL-4Rα subunit bound to a different subunit known as IL-13Rα1. Both type I and type II IL-4Rα receptors are referred to as IL-4 receptors (IL-4R). Type I IL-4Rα is found, for example, on lymphocytes and myeloid cells, and type II IL-4Rα is found, for example, on myeloid and non-hematopoietic cells. These type II receptors have the ability to bind both interleukin-4 and interleukin-13, two cytokines with closely related biological functions.

[0004] Human interleukin-4 (also known as IL-4; BSF-1) is present in a wide range of tissues, including hematopoietic, endothelial, epithelial, muscle, fibroblast, hepatocyte, and brain tissue. Human IL-4 has been found to be an important regulator of humoral and adaptive immunity, playing roles in, for example, stimulating activated B cells, T cell proliferation, B cell differentiation into plasma cells, inducing naive CD4+ T cell differentiation into Th2 effector cells, upregulating MHC class II production, CD23, and IL-4Rα on B cells and myeloid lineage cells (e.g., monocytes), and reducing IL-12 production by Th1 cells, macrophages, IFN-γ, and dendritic cells (Hershey et al., N. Engl. J. Med. 1997, 337(24):1720-5).

[0005] Human interleukin-13 (also known as IL-13; P600) plays an important role in, for example, goblet cell metaplasia, smooth cell muscle contraction, and mucus production in airway epithelia, for example, in allergic asthma. Binding of the IL-13 complex to IL-4Rα initiates activation of multiple transduction pathways, including tyrosine kinase 2 (Tyk-2) and Janus kinase 1 (JAK1). Accordingly, both IL-4 and IL-13 cytokines activate the STAT6 transcription factor, promote B cell class switching to IgE and eosinophil chemotaxis, and have also been found to play potential roles in tumor growth, cell survival, cell adhesion, and metastasis. (Suzuki, A., et al., Cytokine 2015;75(1):79-88). IL-13 is expressed by multiple cell types, including B cells, basophils, eosinophils, mast cells, endothelial cells, fibroblasts, monocytes, macrophages, respiratory epithelial cells, and smooth muscle cells (Hershey et al., N. Engl. J. Med. 1997, 337(24):1720-5).

[0006] Antibody therapeutics targeting type 2 inflammatory diseases are known, either approved, or in clinical development. Such antibodies include dupilumab, which targets IL-4Rα; pascolizumab, which targets IL-4; and lebrikizumab, anrukinzumab, and tralokinumab, which target the IL-13 pathway. (Junttila, I., Frontiers in Immunology 2018, 9:888). However, despite the development of these targeted therapies, there remains a need for additional IL-4Rα antibodies for use in patients with type 2 inflammatory disorders, such as asthma, atopic dermatitis, and / or eosinophilic esophagitis (EoE), as well as in patients who remain resistant to treatment or are non-sustained responders to current therapies, such as dupilumab. DETAILED DESCRIPTION OF THE INVENTION

[0007] The present disclosure provides antibodies that specifically bind to human IL-4Rα and inhibit IL-4R-, IL-4-, and / or IL-13-mediated responses (e.g., B cell proliferation, STAT-6 phosphorylation, CD23 expression), compositions comprising such IL-4Rα antibodies, and methods of using such IL-4Rα antibodies. In particular, the present disclosure provides human IL-4Rα antibodies that bind to a novel epitope spanning the N-terminal fibronectin type III domains 1 and 2 of human and / or cynomolgus monkey IL-4Rα, have desirable binding affinity, block both IL-4- and IL-13-mediated IL-4R signaling, and / or have favorable development properties such as viscosity and / or aggregation. Such human IL-4Rα antibodies can be used to treat type 2 inflammatory disorders associated with IL-4 or IL-13-mediated IL-4R signaling, including atopic dermatitis, eosinophilic esophagitis (EoE), nasal polyps, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU). Such IL-4Rα antibodies can further be used to treat cancer. Thus, the human IL-4Rα antibodies provided herein have one or more of the following properties: 1) bind to a novel structural epitope spanning the N-terminal fibronectin type III domains 1 and 2 of human and / or cynomolgus IL-4Rα, 2) bind to a novel functional epitope on human and / or cynomolgus IL-4Rα, 3) bind to human and / or cynomolgus IL-4Rα with a desired binding affinity, 4) inhibit IL-4-mediated IL-4R signaling, 5) inhibit IL-13-mediated IL-4R signaling, 6) do not significantly induce effector function-mediated killing, 7) do not significantly induce complement fixation, 8) retain Fcγ receptor binding, and / or 9) have low serum protein binding and / or aggregation.

[0008] In some embodiments, an antibody of the present disclosure that specifically binds to human IL-4Rα is a fully human antibody. In some embodiments, an antibody of the present disclosure binds to a novel structural epitope of human IL-4Rα, wherein the epitope spans domain 1 and domain 2 of the n-terminal fibronectin type III domain of IL-4Rα. In some embodiments, an antibody of the present disclosure binds to a novel functional epitope of human IL-4Rα. In some embodiments, an antibody of the present disclosure specifically binds to human IL-4Rα on B cells, T cells, and myeloid cells. In further embodiments, an antibody of the present disclosure binds to human and / or cynomolgus monkey IL-4Rα and blocks the binding of IL-4 and IL-13 to IL-4Rα, thereby preventing IL-4R-mediated signaling. In some embodiments, an antibody of the present disclosure inhibits IL-4- and IL-13-induced IL-4R STAT-6 phosphorylation, B cell proliferation, and CD23 expression. In still further embodiments, the antibodies of the present disclosure do not significantly induce effector function-mediated killing. In some embodiments, anti-human IL-4Rα antibodies having a human IgG4P or human IgG1A backbone retain Fcγ receptor binding. In such embodiments, anti-human IL-4Rα antibodies having a human IgG4P or human IgG1A backbone have improved binding to B cells and myeloid cells compared to human IL-4Rα antibodies having an effector-null backbone (e.g., IgG1AAA).

[0009] In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of human IL-4Rα, wherein the epitope comprises one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L42, L43, E45, H47, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of human IL-4Rα, wherein the epitope includes at least one, at least two, at least three, at least four, or at least five or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L42, L43, E45, H47, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L43, E45, H47, T48, C49, I50, H62, L64, M65, D66, D67, V69, D72, R99, P121, P123, P124, and D125 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of human IL-4Rα, wherein the epitope includes at least one, at least two, at least three, at least four, or at least five or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L43, E45, H47, T48, C49, I50, H62, L64, M65, D66, D67, V69, D72, R99, P121, P123, P124, and D125 (amino acid residue positions correspond to SEQ ID NO: 15).In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising one or more amino acid residues selected from D12, M14, S15, I16, L39, F41, L42, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, a human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of human IL-4Rα, wherein the epitope includes at least one, at least two, at least three, at least four, or at least five or more amino acid residues selected from D12, M14, S15, I16, L39, F41, L42, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In a further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, T48, C49, I50, E52, H62, M65, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In a further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure specifically binds to an epitope of human IL-4Rα, wherein the epitope includes at least one, at least two, at least three, at least four, or at least five or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, T48, C49, I50, E52, H62, M65, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15).In a further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising one or more amino acid residues selected from R99, P121, P123, P124, D125, and P192 (amino acid residue positions corresponding to SEQ ID NO: 15), which are located in domain 2 of the N-terminal fibronectin type III domain of IL-4Rα. In a still further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising at least one or more amino acid residues D66, D67, and D125 (amino acid residue positions corresponding to SEQ ID NO: 15). In a still further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising at least one of amino acid residues D66 and D67 (amino acid residue positions corresponding to SEQ ID NO: 15). In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising at least one of amino acid residues D66 and D125 (amino acid residue position corresponds to SEQ ID NO: 15). In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to an epitope of human IL-4Rα comprising amino acid residue D66 (amino acid residue position corresponds to SEQ ID NO: 15). In yet other embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to a structural and / or functional epitope of human IL-4Rα, which epitope spans domain 1 and domain 2 of the n-terminal fibronectin type III domain of IL-4Rα. In certain embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof specifically binds to a novel structural and / or functional epitope of human IL-4Rα, wherein the epitope overlaps with the IL-4-binding site for IL-4Rα. In such embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof blocks the binding of IL-4 to human IL-4Rα. In certain embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof binds to a novel structural and / or functional epitope on human IL-4Rα, where the epitope overlaps with the IL-13-binding site on IL-4Rα.In such embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof blocks IL-13 binding to human IL-4Rα. In certain embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof binds to a novel structural and / or functional epitope of human IL-4Rα, where the epitope overlaps with both the IL-4-binding site and the IL-13-binding site on IL-4Rα. In such embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof blocks IL-4 and IL-13 binding to human IL-4Rα. In some embodiments, the IL-4Rα epitope is determined by X-ray crystallography, alanine scanning mutagenesis, steric hindrance mutagenesis, and / or HDX-MS. In yet other embodiments, the IL-4Rα epitope is determined by site-directed mutagenesis.

[0010] In some embodiments, the present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα and comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises SEQ ID NO: 1, HCDR2 comprises SEQ ID NO: 2, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 4, LCDR2 comprises SEQ ID NO: 5, and LCDR3 comprises SEQ ID NO: 6. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα comprises a VH comprising SEQ ID NO: 7 and a VL comprising SEQ ID NO: 8. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC comprising SEQ ID NO: 33 and an LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC comprising SEQ ID NO: 35 and an LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises an HC comprising SEQ ID NO: 9 and an LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 13 and a LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 37 and a LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 31 and a LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 50 and a LC comprising SEQ ID NO: 10. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 52 and a LC comprising SEQ ID NO: 10.

[0011] In some embodiments, the disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity-determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises SEQ ID NO: 42, HCDR2 comprises SEQ ID NO: 2, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 22, LCDR2 comprises SEQ ID NO: 5, and LCDR3 comprises SEQ ID NO: 6. In some embodiments, the antibody or antigen-binding fragment thereof that specifically binds to IL-4Rα comprises a VH comprising SEQ ID NO: 44 and a VL comprising SEQ ID NO: 45. In some embodiments, the antibody or antigen-binding fragment thereof comprises a HC comprising SEQ ID NO: 46 and a LC comprising SEQ ID NO: 47.

[0012] In some embodiments, the present disclosure provides an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, wherein the antibody or antigen-binding fragment comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity-determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity-determining regions LCDR1, LCDR2, and LCDR3, wherein HCDR1 comprises SEQ ID NO: 19, HCDR2 comprises SEQ ID NO: 20, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 22, LCDR2 comprises SEQ ID NO: 5, and LCDR3 comprises SEQ ID NO: 24. In some embodiments, the human IL-4Rα antibody comprises a VH comprising SEQ ID NO: 25 and a VL comprising SEQ ID NO: 26. In some embodiments, the antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα comprises a heavy chain (HC) comprising SEQ ID NO: 27 and a light chain (LC) comprising SEQ ID NO: 28.

[0013] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα has a human IgG1 or human IgG4 isotype.

[0014] In some embodiments of the present disclosure, an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα has a human IgG1 isotype. In some embodiments, the human IL-4Rα antibody has a modified human IgG1 Fc region, wherein the antibody comprises an alanine at amino acid residue 322 (K322A substitution), also referred to as IgG1A (EU numbering). In such embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure has reduced or eliminated complement activity. In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG1 Fc region comprising L234A, L235A, and / or P329A, also referred to as IgG1AA or IgG1AAA, respectively, that reduces or eliminates binding to Fcγ and C1q receptors (all residues numbered according to EU numbering). In some embodiments, human IL-4Rα antibodies or antigen-binding fragments thereof having a human IgG1A backbone exhibit improved binding to B cells and myeloid cells when compared to human IL-4Rα antibodies having a human IgG1AAA effector-null backbone.

[0015] In some embodiments of the present disclosure, the antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα has a human IgG4 isotype. In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 hinge region containing a S228P substitution (EU numbering), also known as IgG4P, which reduces IgG4 Fab arm exchange in vivo (see Labrijn, et al., Nat. Biotechnol. 2009, 27(8):767). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 Fc region containing F234A and / or L235A (EU numbering), also known as IgG4AA, which reduces binding to Fcγ and C1q receptors. According to some embodiments of the present disclosure, the Fc region contains S228P, F234A, and L235A (all residues numbered according to IMGT or EU numbering), also known as IgG4PAA. In some embodiments, a human IL-4Rα antibody or antigen-binding fragment thereof having a human IgG4P backbone has improved binding to B cells and myeloid cells compared to a human IL-4Rα antibody having an effector-null backbone.

[0016] In some embodiments, a human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC region that reduces the viscosity of the antibody compared to the same antibody having a wild-type human IgG4 HC constant region. In such embodiments, a human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region that includes an amino acid substitution at any one or more of the following amino acid residues relative to the wild-type human IgG4 HC constant region: Q274K, Q355R, E419Q (all positions numbered according to EU numbering).

[0017] In other embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC region that includes an amino acid substitution at any one or more of the following amino acid residues relative to the wild-type human IgG4 HC region: E137G, D203N, Q274K, Q355R, E419Q (all positions numbered according to EU numbering).

[0018] In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising an E137G substitution (EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising a D203N substitution (EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising a Q274K substitution (EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising a Q355R substitution (EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising an E419Q substitution (EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising Q274K and Q355R substitutions (all positions are numbered according to EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising Q274K and E419Q substitutions (all positions are numbered according to EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising Q355R and E419Q substitutions (all positions are numbered according to EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising Q274K, Q355R, and E419Q substitutions (all positions are numbered according to EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC constant region comprising S228P, Q274K, Q355R, E419Q substitutions (all positions numbered according to EU numbering). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG4 HC region comprising S228P, E137G, D203N, Q274K, Q355R, E419Q substitutions (all positions numbered according to EU numbering).

[0019] In some embodiments, the antibody or antigen-binding fragment thereof comprises one or more of the following: a glycine at amino acid residue 137 (EU numbering), an asparagine at amino acid residue 203 (EU numbering), a lysine at amino acid residue 274 (EU numbering), an arginine at amino acid residue 355 (EU numbering), or a glutamine at amino acid residue 419 (EU numbering).

[0020] In some embodiments, the antibody or antigen-binding fragment thereof comprises a lysine at amino acid residue 274 (EU numbering), an arginine at amino acid residue 355, and a glutamine at amino acid residue 419 (EU numbering).

[0021] In some embodiments, the antibody or antigen-binding fragment thereof comprises a glycine at amino acid residue 137 (EU numbering), an asparagine at amino acid residue 203 (EU numbering), a lysine at amino acid residue 274 (EU numbering), an arginine at amino acid residue 355, and a glutamine at amino acid residue 419 (EU numbering).

[0022] In further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof has a modified human IgG1 or human IgG4 constant domain containing an engineered cysteine ​​residue for use in generating antibody conjugate compounds (also called bioconjugates) (see WO 2018 / 232088). More particularly, in such embodiments of the present disclosure, the human IL-4Rα antibody or antigen-binding fragment thereof contains a cysteine ​​at amino acid residue 124 (EU numbering), or a cysteine ​​at amino acid residue 378 (EU numbering), or a cysteine ​​at amino acid residue 124 (EU numbering) and at amino acid residue 378 (EU numbering).

[0023] In some embodiments, the antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα has modified human IgG1 or human IgG4 framework regions. In some embodiments, the modifications are in the VH framework regions. In some embodiments, the modifications are in the VL framework regions. In some embodiments, the modifications are in the VH and VL framework regions. In further embodiments, the modified human IgG1 or human IgG4 framework regions reduce the immunogenic risk of the antibody.

[0024] In some embodiments of the present disclosure, a human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure binds to IL-4Rα and inhibits the binding of human IL-4 and human IL-13 to human IL-4Rα. In such embodiments, the antibody or antigen-binding fragment thereof of the present disclosure inhibits the binding of human IL-4 to human IL-4Rα, thereby inhibiting human IL-4R activation, STAT-6 phosphorylation, B cell and / or T cell proliferation, and CD23 expression. In such embodiments, the antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα and inhibits the binding of human IL-4 to human IL-4Rα by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. In further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα on B cells and T cells and inhibits IL-4-induced STAT-6 phosphorylation in B cells and T cells by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. In further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα on B cells and inhibits IL-4-induced B cell proliferation by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. In still other embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα on myeloid cells and inhibits IL-4-induced CD23 expression on human myeloid cells by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%.

[0025] In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure inhibits the binding of human IL-13 to human IL-4Rα. In such embodiments, the antibody or antigen-binding fragment thereof of the present disclosure inhibits the binding of human IL-13 to human IL-4Rα, and thus inhibits human IL-4R activation, STAT-6 phosphorylation, B cell and / or T cell proliferation, and CD23 expression. In further embodiments, the antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα and inhibits the binding of human IL-13 to human IL-4Rα by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure binds to human IL-4Rα on B cells and inhibits IL-13-induced STAT-6 activation in B cells by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. In further embodiments, the antibody or antigen-binding fragment thereof of the present disclosure inhibits binding of human IL-13 to human IL-4Rα and inhibits B cell and / or T cell proliferation. In such embodiments, the human IL-4Rα antibodies or antigen-binding fragments thereof of the present disclosure bind to human IL-4Rα on B cells and / or T cells and inhibit IL-13-induced B cell and / or T cell proliferation by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In yet other embodiments, the human IL-4Rα antibodies or antigen-binding fragments thereof of the present disclosure bind to human IL-4Rα on myeloid cells and inhibit IL-13-induced CD23 expression on human myeloid cells by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0026] In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof of the present disclosure inhibits the binding of human IL-4 and human IL-13 to human IL-4Rα by binding to a novel epitope in human IL-4Rα, wherein the epitope includes one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L42, L43, E45, H47, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, wherein the epitope comprises one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, F41, L43, E45, H47, T48, C49, I50, H62, L64, M65, D66, D67, V69, D72, R99, P121, P123, P124, and D125 (amino acid residue positions correspond to SEQ ID NO: 15). In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, wherein the epitope includes one or more amino acid residues selected from D12, M14, S15, I16, L39, F41, L42, T48, C49, I50, E52, H62, L64, M65, D66, D67, V68, V69, D72, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15).In some embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, wherein the epitope includes one or more amino acid residues selected from D12, M14, S15, I16, Y37, L39, T48, C49, I50, E52, H62, M65, R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15). In a further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope in human IL-4Rα, wherein the epitope comprises one or more amino acid residues selected from R99, P121, P123, P124, D125, and P192 (amino acid residue positions correspond to SEQ ID NO: 15), which are located in domain 2 of the N-terminal fibronectin type III domain of IL-4Rα. In yet a further embodiment, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope in human IL-4Rα, wherein the epitope comprises at least one of amino acid residues D66, D67, and D125 (amino acid residue positions correspond to SEQ ID NO: 15). In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, the epitope comprising at least one of amino acid residues D66 and D67 (amino acid residue positions corresponding to SEQ ID NO: 15). In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, the epitope comprising at least one of amino acid residues D66 and D125 (amino acid residue positions corresponding to SEQ ID NO: 15).In still further embodiments, the human IL-4Rα antibody or antigen-binding fragment thereof inhibits binding of human IL-4 and / or human IL-13 to human IL-4Rα by binding to an epitope on human IL-4Rα, wherein the epitope includes amino acid residue D66 (amino acid residue position corresponds to SEQ ID NO: 15).

[0027] Some embodiments of the present disclosure provide nucleic acids encoding the heavy or light chain, or VH or VL, of a novel antibody that specifically binds to human IL-4Rα, or a vector comprising such a nucleic acid.

[0028] In some embodiments, the disclosure provides a nucleic acid comprising the sequence of SEQ ID NO: 11, 12, 14, 29, 30, 32, 34, 36, 38, 48, 49, 51, or 53.

[0029] In some embodiments, nucleic acids encoding the heavy or light chain of an antibody that specifically binds to human IL-4Rα are provided. In some embodiments, nucleic acids comprising a sequence encoding SEQ ID NO: 9, 10, 13, 27, 28, 31, 33, 35, 37, 46, 47, 50, or 52 are provided. In some embodiments, nucleic acids comprising a sequence encoding an antibody heavy chain comprising SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52 are provided. For example, the nucleic acid may comprise a sequence selected from SEQ ID NO: 11, 14, 29, 32, 34, 36, 38, 48, 51, or 53. In some embodiments, nucleic acids comprising a sequence encoding an antibody light chain comprising SEQ ID NO: 10, 28, or 47 are provided. For example, the nucleic acid may comprise a sequence selected from SEQ ID NO: 12, 30, or 49.

[0030] In some embodiments of the present disclosure, nucleic acids are provided that encode the VH or VL of an antibody that specifically binds to human IL-4Rα. In some embodiments, nucleic acids are provided that comprise a sequence encoding SEQ ID NO: 7, 8, 25, 26, 44, or 45. In some embodiments, nucleic acids are provided that comprise a sequence encoding the antibody VH comprising SEQ ID NO: 7, 25, or 44. In some embodiments, nucleic acids are provided that comprise a sequence encoding the antibody VL comprising SEQ ID NO: 8, 26, or 45.

[0031] In some embodiments of the present disclosure, a vector is provided comprising a nucleic acid sequence encoding an antibody heavy or light chain. For example, such a vector can comprise a nucleic acid sequence encoding SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52. In some embodiments, the vector comprises a nucleic acid sequence encoding SEQ ID NO: 10, 28, or 47.

[0032] Also provided herein are vectors comprising a nucleic acid sequence encoding an antibody VH or VL. For example, such vectors can comprise a nucleic acid sequence encoding SEQ ID NO: 7, 25, or 44. In some embodiments, the vector comprises a nucleic acid sequence encoding SEQ ID NO: 8, 26, or 45.

[0033] Also provided herein are vectors comprising a first nucleic acid sequence encoding an antibody heavy chain and a second nucleic acid sequence encoding an antibody light chain. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52 and a second nucleic acid sequence encoding SEQ ID NO: 10, 28, or 47.

[0034] In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:9 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:13 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:27 and a second nucleic acid sequence encoding SEQ ID NO:28. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:31 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:33 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:35 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:37 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence encoding SEQ ID NO:50 and a second nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the vector comprises a first nucleic acid sequence that encodes SEQ ID NO: 52 and a second nucleic acid sequence that encodes SEQ ID NO: 10. In some embodiments, the vector comprises a first nucleic acid sequence that encodes SEQ ID NO: 46 and a second nucleic acid sequence that encodes SEQ ID NO: 47.

[0035] Also provided are compositions comprising a first vector comprising a nucleic acid sequence encoding an antibody heavy chain and a second vector comprising a nucleic acid sequence encoding an antibody light chain, hi some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52 and a second nucleic acid sequence encoding SEQ ID NO: 10, 28, or 47.

[0036] In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:9 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:13 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:27 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:28. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:31 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:33 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:35 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO:37 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 50 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 52 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10. In some embodiments, the composition comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 46 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 47.

[0037] The nucleic acids of the present disclosure can be expressed in host cells, for example, after the nucleic acid is operably linked to an expression control sequence. Expression control sequences capable of expressing an operably linked nucleic acid are well known in the art. The expression vector may include a sequence encoding one or more signal peptides that facilitate secretion of the polypeptide from the host cell. The expression vector containing the nucleic acid of interest (e.g., a nucleic acid encoding an antibody heavy or light chain) can be transferred into a host cell by well-known methods, such as stable or transient transfection, transformation, transduction, or infection. Additionally, the expression vector may include one or more selectable markers, such as tetracycline, neomycin, and dihydrofolate reductase, to facilitate detection of host cells transformed with the desired nucleic acid sequence.

[0038] In another aspect, provided herein are cells, e.g., host cells, comprising a nucleic acid, vector, or nucleic acid composition described herein. Host cells can be cells stably or transiently transfected, transformed, transduced, or infected with one or more expression vectors expressing all or a portion of an antibody described herein. In some embodiments, host cells can be stably or transiently transfected, transformed, transduced, or infected with expression vectors expressing the HC and LC polypeptides of an antibody of the present disclosure. In some embodiments, host cells can be stably or transiently transfected, transformed, transduced, or infected with a first vector expressing the HC polypeptide and a second vector expressing the LC polypeptide of an antibody described herein. Such host cells, e.g., mammalian host cells, can express antibodies that specifically bind to human IL-4Rα described herein. Mammalian host cells known to be capable of expressing antibodies include CHO cells, HEK293 cells, COS cells, and NS0 cells.

[0039] In some embodiments, a cell, e.g., a host cell, comprises a vector comprising a first nucleic acid sequence encoding SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52 and a second nucleic acid sequence encoding SEQ ID NO: 10, 28, or 47.

[0040] In some embodiments, a cell, e.g., a host cell, comprises a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, 13, 27, 31, 33, 35, 37, 46, 50, or 52, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10, 28, or 47.

[0041] The present disclosure further provides a process for producing an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα described herein by culturing the above-described host cells, e.g., mammalian host cells, under conditions such that the antibody is expressed, and recovering the expressed antibody from the culture medium. The medium into which the antibody is secreted can be purified by conventional techniques. Various methods of protein purification can be employed, and such methods are known in the art and are described, for example, in Deutscher, Methods in Enzymology 182:83-89 (1990), and Scopes, Protein Purification: Principles and Practice, 3rd Edition, Springer, NY (1994).

[0042] The present disclosure further provides an antibody or antigen-binding fragment thereof produced by any of the processes described herein.

[0043] In another aspect, the present invention provides pharmaceutical compositions comprising the antibodies, nucleic acids, or vectors described herein. Such pharmaceutical compositions may also comprise one or more pharmaceutically acceptable excipients, diluents, or carriers. Pharmaceutical compositions can be prepared by methods well known in the art (e.g., Remington: The Science and Practice of Pharmacy, 22nd ed. (2012), A. Loyd et al., Pharmaceutical Press).

[0044] The antibodies or antigen-binding fragments thereof, nucleic acids, vectors, or pharmaceutical compositions that specifically bind to human IL-4Rα described herein can be used to treat IL-4R-associated disorders, e.g., immunoinflammatory disorders, e.g., type 2 inflammatory disorders (including, but not limited to, atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU)). Accordingly, the antibodies of the present disclosure can further be used to treat cancer, e.g., B-cell-related cancers, e.g., CLL, or other cancers, e.g., malignant glioma, ovarian cancer, lung cancer, breast cancer, squamous cell carcinoma of the head and neck (SCCHN), pancreatic cancer, kidney cancer, colon cancer, prostate cancer, and bladder cancer. Such methods may further comprise administering one or more chemotherapeutic agents to the subject. In some embodiments, the chemotherapeutic agent is administered in simultaneous, separate, or sequential combination with the antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα or a pharmaceutical composition thereof described herein.Embodiments of the present disclosure further provide methods of treating cancer using an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα or a pharmaceutical composition thereof in simultaneous, separate, or sequential combination with ionizing radiation.

[0045] In some embodiments, provided herein are methods for treating an IL-4R-associated disorder, e.g., an immunoinflammatory disorder such as a type 2 immunoinflammatory disorder, or cancer, in a subject (e.g., a human patient) in need thereof by administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that specifically binds human IL-4Rα, a nucleic acid encoding such an antibody that specifically binds human IL-4Rα, a vector comprising such a nucleic acid, or a pharmaceutical composition comprising such an antibody, nucleic acid, or vector that specifically binds human IL-4Rα described herein. The antibody, nucleic acid, vector, or pharmaceutical composition described herein can be administered parenterally (e.g., subcutaneously and intravenously). In embodiments, the IL-4Rα / IL-4 and / or IL-4Rα / IL-13-associated immunoinflammatory disorder is a type 2 inflammatory disorder. Such type 2 inflammatory disorders include, but are not limited to, atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU). In some embodiments, the order of IL-4R is cancer, e.g., B-cell related cancer, e.g., CLL, or other cancers, e.g., malignant glioma, ovarian cancer, lung cancer, breast cancer, squamous cell carcinoma of the head and neck (SCCHN), pancreatic cancer, kidney cancer, colon cancer, prostate cancer, and bladder cancer.

[0046] Also provided herein are antibodies or antigen-binding fragments thereof, nucleic acids, vectors, or pharmaceutical compositions described herein that specifically bind to human IL-4Rα for use in therapy. Furthermore, the present disclosure also provides antibodies or antigen-binding fragments thereof that specifically bind to human IL-4Rα, nucleic acids, vectors, or pharmaceutical compositions described herein for use in treating IL-4R disorders, e.g., immunoinflammatory disorders such as type 2 inflammatory disorders, or cancer. Such type 2 inflammatory disorders include, but are not limited to, atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU). In some embodiments, the IL-4R-associated disorder is cancer, e.g., a B-cell-associated cancer, e.g., CLL, or other cancers, e.g., malignant glioma, ovarian cancer, lung cancer, breast cancer, squamous cell carcinoma of the head and neck (SCCHN), pancreatic cancer, kidney cancer, colon cancer, prostate cancer, and bladder cancer.

[0047] Provided herein is the use of an antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, a nucleic acid, a vector, or a pharmaceutical composition described herein in the manufacture of a medicament for treating an IL-4R-associated disorder, e.g., an immunoinflammatory disorder such as a type 2 inflammatory disorder or cancer. Such type 2 inflammatory disorders include, but are not limited to, atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU). In some embodiments, the IL-4R-associated disorder is cancer, e.g., a B-cell-related cancer, e.g., CLL, or other cancers, e.g., malignant glioma, ovarian cancer, lung cancer, breast cancer, squamous cell carcinoma of the head and neck (SCCHN), pancreatic cancer, kidney cancer, colon cancer, prostate cancer, and bladder cancer.

[0048] As used herein, the term "IL-4Rα," unless otherwise specified, refers to any naturally occurring mature IL-4Rα resulting from intracellular processing of an IL-4Rα precursor protein. The term includes IL-4Rα from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus or rhesus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of IL-4Rα, such as splice variants or allelic variants. The amino acid sequence of an exemplary human IL-4Rα is known in the art, e.g., UniProt Reference Sequence P24394 (SEQ ID NO: 39). The amino acid sequence of an exemplary cynomolgus IL-4Rα is also known in the art, e.g., NCBI Reference No. XP_005591572.2 (SEQ ID NO: 40). The term "IL-4Rα" is used herein to collectively refer to all known human IL-4Rα isoforms and polymorphic forms. Sequencing as used herein is based on the mature protein, without the signal peptide.

[0049] As used herein, unless otherwise specified, the term "IL-4R" refers to the complex of the IL-4R α subunit and the common γ chain (type I receptor) or the complex of the IL-4R α subunit and IL-13Rα1 (type II receptor).

[0050] The term "IL-4," as used herein, unless otherwise specified, refers to any naturally occurring mature IL-4 resulting from intracellular processing of an IL-4 precursor protein. The term includes IL-4 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus or rhesus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of IL-4, such as splice variants or allelic variants. The amino acid sequence of an example of human IL-4, e.g., UniProt Reference Sequence P05112 (SEQ ID NO: 17), is known in the art. The term "IL-4" is used herein to collectively refer to all known human IL-4 isoforms and polymorphic forms.

[0051] The term "IL-13," as used herein, unless otherwise specified, refers to any naturally occurring mature IL-13 resulting from intracellular processing of an IL-13 precursor protein. The term includes IL-13 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus or rhesus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of IL-13, such as splice variants or allelic variants. An exemplary amino acid sequence of human IL-13, e.g., UniProt Reference Sequence P35225 (SEQ ID NO: 18), is known in the art. The term "IL-13" is used herein to collectively refer to all known human IL-13 isoforms and polymorphic forms.

[0052] The term "CD23," as used herein, unless otherwise specified, refers to any naturally occurring mature CD23 resulting from intracellular processing of a CD23 precursor protein. The term includes CD23 from any vertebrate source, including mammals such as primates (e.g., humans and cynomolgus or rhesus monkeys) and rodents (e.g., mice and rats), unless otherwise indicated. The term also includes naturally occurring variants of CD23, such as splice variants or allelic variants. The amino acid sequence of one example of human CD23 is known in the art, e.g., UniProt Reference SEQ ID NO: P06734 (SEQ ID NO: 41). The term "CD23" is used herein to collectively refer to all known human CD23 isoforms and polymorphic forms.

[0053] As used herein, the term "IL-4R-associated disorder" refers to a disorder associated with IL-4R-mediated signaling, such as, for example, disorders associated with type 1 IL-4R and type II IL-4R signaling. Such IL-4R-associated disorders can include, for example, immunoinflammatory disorders. Such immunoinflammatory disorders can include type 2 inflammatory disorders as disclosed herein. IL-4R-associated disorders can further include cancer.

[0054] As used herein, the term "antibody" refers to an immunoglobulin molecule that binds to an antigen. Antibody embodiments include monoclonal antibodies, polyclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, bispecific or multispecific antibodies, or conjugated antibodies. The antibody may be of any class (e.g., IgG, IgE, IgM, IgD, IgA) and any subclass (e.g., IgG1, IgG2, IgG3, IgG4).

[0055] An exemplary antibody is an immunoglobulin G (IgG) antibody composed of four polypeptide chains: two heavy chains (HC) and two light chains (LC) cross-linked via interchain disulfide bonds. The amino-terminal portion of each of the four polypeptide chains contains a variable region of approximately 100 to 125 amino acids primarily responsible for antigen recognition. The carboxy-terminal portion of each of the four polypeptide chains contains a constant region primarily responsible for effector function. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region refers to the region of an antibody that contains the Fc region and CH1 domain of the antibody heavy chain. Each light chain is composed of a light chain variable region (VL) and a light chain constant region. IgG isotypes can be further divided into subclasses (e.g., IgG1, IgG2, IgG3, and IgG4). The numbering of amino acid residues in the constant region is based on the EU index as in Kabat. Kabat et al, Sequences of Proteins of Immunological Interest, 5th edition, Bethesda, MD: US Department of Health and Human Services, Public Health Service, National Institutes of Health (1991). The terms EU index numbering or EU numbering are used interchangeably herein.

[0056] The VH and VL regions can be further subdivided into hypervariable regions, called complementarity-determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). CDRs are exposed on the surface of the protein and are critical regions of the antibody for antigen-binding specificity. Each VH and VL is composed of three CDRs and four FRs, arranged from the amino terminus to the carboxy terminus in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Herein, the three CDRs of the heavy chain are referred to as "HCDR1, HCDR2, and HCDR3," and the three CDRs of the light chain are referred to as "LCDR1, LCDR2, and LCDR3." CDRs contain most of the residues that form specific interactions with the antigen.The assignment of amino acid residues to CDRs can be performed using the methods of Kabat (Kabat et al., "Sequences of Proteins of Immunological Interest", National Institutes of Health, Bethesda, Md. (1991)), Chothia (Chothia et al., "Canonical structures for the hypervariable regions of immunoglobulins", Journal of Molecular Biology, 196, 901-917 (1987), Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927-948 (1997)), North (North et al., "A New Clustering of Antibody CDR Loop Conformations", Journal of Molecular Biology, 406, 228-256 (2011)), or IMGT (the international ImMunoGeneTics The North CDR definitions can be used for antibodies that specifically bind to human IL-4Rα described herein.

[0057] Embodiments of the present disclosure also include antibody fragments or antigen-binding fragments that comprise at least a portion of an antibody that retains the ability to specifically interact with an antigen, such as, for example, Fab, Fab', F(ab')2, Fv fragments, scFv, scFab, disulfide-linked Fv (sdFv), Fd fragments, and linear antibodies, which may be fused to an Fc region or an IgG heavy chain constant region.

[0058] It is fused to an Fc region or an IgG heavy chain constant region.

[0059] As used herein, the term "Fc region" refers to the region of an antibody comprising the CH2 and CH3 domains of the antibody heavy chain. Optionally, the Fc region may include a portion or the entire hinge region of the antibody heavy chain. Biological activities, such as effector functions, are attributed to the Fc region, which vary depending on the antibody isotype. Examples of antibody effector functions include Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), C1q binding, complement-dependent cytotoxicity (CDC), phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and B cell activation.

[0060] The term "epitope" as used herein refers to the amino acid residues of an antigen that are bound by an antibody. An epitope can be a linear epitope, a conformational epitope, or a hybrid epitope. The term "epitope" can be used in reference to a structural epitope. A structural epitope, according to some embodiments, can be used to describe the region of an antigen that is covered by an antibody (e.g., the footprint of an antibody when bound to the antigen). In some embodiments, a structural epitope can describe amino acid residues of an antigen that are within a specific proximity (e.g., within a specific number of angstroms) of amino acid residues of an antibody. The term "epitope" can also be used in reference to a functional epitope. A functional epitope, according to some embodiments, can be used to describe amino acid residues of an antigen that interact with amino acid residues of an antibody in a manner that contributes to the binding energy between the antigen and the antibody. Epitopes can be determined according to different experimental techniques, also referred to as "epitope mapping techniques." It is understood that the determination of an epitope may vary based on the different epitope mapping techniques used, and may also vary depending on the different experimental conditions used, for example, due to conformational changes or cleavage of the antigen induced by the specific experimental conditions. Epitope mapping techniques are known in the art (e.g., Rockberg and Nilvebrant, Epitope Mapping Protocols: Methods in Molecular Biology, Humana Press, 3 rd ed. 2018; Holst et al., Molecular Pharmacology 1998, 53(1):166-175), for example, X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, site-directed mutagenesis, species-exchange mutagenesis, alanine scanning mutagenesis, steric hindrance mutagenesis, hydrogen-deuterium exchange (HDX), and cross-blocking assays.

[0061] The term "bind," as used herein, unless otherwise specified, is intended to mean the ability of a protein or molecule to form a chemical bond or attractive interaction with another protein or molecule, resulting in proximity of the two proteins or molecules as determined by common methods known in the art.

[0062] The term "nucleic acid," as used interchangeably herein, refers to a polymer of nucleotides, including single- and / or double-stranded nucleotide-containing molecules, such as DNA, cDNA, and RNA molecules, that incorporate naturally occurring nucleotides, modified nucleotides, and / or nucleotide analogs. A polynucleotide of the present disclosure can also include substrates incorporated therein, for example, by a DNA or RNA polymerase or a synthetic reaction.

[0063] As used herein, the term "subject" refers to a mammal, including, but not limited to, humans, chimpanzees, apes, monkeys, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rats, mice, guinea pigs, etc. Preferably, the subject is a human.

[0064] The term "therapeutically effective amount" as used herein refers to an amount of a protein or nucleic acid or vector or composition that, upon single or multiple administration to a subject, provides a desired effect in a subject undergoing diagnosis or treatment. The term "therapeutically effective amount" as used herein further refers to an amount or dose of a protein or nucleic acid or vector or composition of the present disclosure that elicits a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity, or improving symptoms, alleviating a condition, slowing or delaying disease progression, or preventing a disease. In non-limiting embodiments, the term "therapeutically effective amount" refers to an amount (in terms of dosage, duration, and means of administration) of a protein or nucleic acid or vector or composition that, when administered to a subject, is effective to at least partially alleviate, inhibit, prevent, and / or ameliorate a condition, disorder, or disease. The effective amount of a protein or antibody or vector or composition may vary depending on factors such as the individual's condition, age, sex, and weight, as well as the ability of the protein or antibody or vector or composition to elicit a desired response in the individual. An effective amount is also an amount in which any toxic or adverse effects of the protein or nucleic acid or vector or composition of the present invention are outweighed by the therapeutically beneficial effects.

[0065] The term "inhibit" as used herein refers to, for example, decreasing, reducing, slowing, diminishing, stopping, disrupting, eliminating, antagonizing, or blocking a biological response or activity, but does not necessarily indicate complete elimination of the biological response.

[0066] As used herein, "treatment" or "treating" refers to any process that may slow, control, retard, or halt the progression of a disorder or disease symptom disclosed herein, but does not necessarily indicate the complete disappearance of all disorder or disease symptoms. Treatment includes the administration of a protein or nucleic acid or vector or composition for the treatment of a disease or condition in a patient, particularly a human.

[0067] As used herein, the term "about" means within 5%.

[0068] As used herein, the terms "a," "an," "the," and similar terms as used in the context of this disclosure (particularly in the context of the claims) should be construed to encompass both the singular and the plural, unless otherwise specified herein or clearly contradicted by context.

[0069] A "chemotherapeutic agent" or "chemotherapeutic agent," as used interchangeably herein, is a chemical agent or drug that selectively destroys cancer cells and tissues. Chemotherapeutic agents may include, but are not limited to, compounds such as taxane compounds, compounds that act via a taxane mechanism, platinum compounds, anthracycline compounds, antimetabolites, alkylating agents, epipodophyllotoxin compounds, camptothecin compounds, topoisomerase inhibitors, mitotic inhibitors, or any combination thereof. Chemotherapeutic agents can be administered alone or in combination with other therapeutic agents.

[0070] As used herein, the term "ionizing radiation" refers to radiation of a specific wavelength that is used to destroy or damage cancer cells. Ionizing radiation includes radon, X-rays, gamma rays, and other forms of high-energy radiation. Ionizing radiation can include external radiation (or external beam radiation), internal radiation (or brachytherapy), or whole-body radiation. [Brief explanation of the drawings]

[0071] [Figure 1] 1 shows an X-ray crystal structure overlay of the Fab portion of 8660 human IL-4Rα antibody bound to the IL-4Rα ECD and the crystal structure of the dupilumab Fab portion with Crystal Kappa design complexed with human IL-4Rα (pdb accession code 6 WGL). [Figure 2] The functional epitope amino acid residue positions (human IL-4Rα residues Asp66 and Asp125) are shown in the crystal structure of the 5F3 Fab portion with Crystal Kappa design complexed with human IL-4Rα ECD. [Figure 3A]Figure 3 shows the binding specificity of the 5F3 and 5559 human IL-4Rα antibodies to human IL-4Rα (3A) and cynomolgus monkey IL-4Rα (3B) as measured by MSD ELISA assay. [Figure 3B] Figure 3 shows the binding specificity of the 5F3 and 5559 human IL-4Rα antibodies to human IL-4Rα (3A) and cynomolgus monkey IL-4Rα (3B) as measured by MSD ELISA assay. [Figure 4A] 4A and 4C show the binding specificity of the 5559 human IL-4Rα antibody to IL-4Rα on human B cells (4A and 4C) and T cells (4B). [Figure 4B] 4A and 4C show the binding specificity of the 5559 human IL-4Rα antibody to IL-4Rα on human B cells (4A and 4C) and T cells (4B). [Figure 4C] 4A and 4C show the binding specificity of the 5559 human IL-4Rα antibody to IL-4Rα on human B cells (4A and 4C) and T cells (4B). [Figure 5A] Figure 1 shows that the 5F3, 8660 and 5559 human IL-4Rα antibodies block the binding of IL-4 (5A) and IL-13 (5B) to IL-4Rα in a cell-based assay. [Figure 5B] Figure 1 shows that the 5F3, 8660 and 5559 human IL-4Rα antibodies block the binding of IL-4 (5A) and IL-13 (5B) to IL-4Rα in a cell-based assay. [Figure 6A] Figure 6 shows that the 5559 human IL-4Rα antibody inhibits IL-4-induced pSTAT6 phosphorylation in primary human B cells (6B) and primary human T cells (6A). [Figure 6B] Figure 6 shows that the 5559 human IL-4Rα antibody inhibits IL-4-induced pSTAT6 phosphorylation in primary human B cells (6B) and primary human T cells (6A). [Figure 6C] show that the 5559 human IL-4Rα antibody inhibits IL-13-induced pSTAT6 phosphorylation in human B cells. [Figure 7]Figure 1 shows that the 5559 human IL-4Rα antibody inhibits IL-4-induced B cell proliferation. [Figure 8A] Figure 1 shows that the 5559 human IL-4Rα antibody inhibits IL-4 (8A) and IL-13 (8B)-induced CD23 expression in myeloid cells. [Figure 8B] Figure 1 shows that the 5559 human IL-4Rα antibody inhibits IL-4 (8A) and IL-13 (8B)-induced CD23 expression in myeloid cells. [Figure 9] 5 shows that the 5559 human IL-4Rα antibody does not bind to complement component C1q in an ELISA assay. [Figure 10A] 10A and 10B show that the 5559 human IL-4Rα antibody does not significantly induce ADCC activity in either a reporter gene-based assay (10A) or a primary cell-based assay (10B). [Figure 10B] 10A and 10B show that the 5559 human IL-4Rα antibody does not significantly induce ADCC activity in either a reporter gene-based assay (10A) or a primary cell-based assay (10B). [Figure 11] 5 shows that the 5559 human IL-4Rα antibody does not induce CDC activity in Daudi cells. [Figure 12A] 1 shows a differential scanning calorimetry (DSC) thermogram of the 5559 human IL-4Rα antibody. [Figure 12B] 1 shows a differential scanning calorimetry (DSC) thermogram of the 5559 human IL-4Rα antibody. [Figure 13] 1 shows an X-ray crystal structure overlay of the Fab portion of the 5559 human IL-4Rα antibody bound to the IL-4Rα ECD and the crystal structure of the dupilumab Fab portion with Crystal Kappa design complexed with human IL-4Rα (pdb accession code 6 WGL). [Figure 14]The crystal structure of the 5559 Fab portion with the crystal kappa design complexed with the human IL-4Rα ECD shows the amino acid residue positions Asp66, Asp67, and Asp125 of human IL-4Rα (all identified in the structural epitope, and additionally Asp66 in the functional epitope). [Example]

[0072] Example 1: Generation and manipulation of antibodies that bind to human IL-4Rα (anti-human IL-4Rα antibodies) Antibody generation: To develop antibodies specific to human IL-4Rα, transgenic mice carrying human immunoglobulin variable regions were immunized with the Fc-tagged extracellular domain (ECD) of human IL-4Rα and alternately boosted with human and cynomolgus monkey Fc-tagged IL-4Rα ECD proteins. Screening was performed using histidine-tagged human and cynomolgus monkey IL-4Rα ECDs to identify cross-reactivity, and in the absence or presence of excess soluble IL-4 to identify IL-4-blocking antibodies. Cross-reactive antibodies were cloned as Fabs, expressed, and purified by standard procedures and tested for blocking activity against IL-4 and IL-13 in a reporter cell line, human embryonic kidney (HEK)-Blue IL-4 / IL-13 (InvivoGen). Antibodies were selected and their CDRs, variable domain framework regions, and IgG isotype were engineered to improve properties such as affinity, stability, solubility, viscosity, hydrophobicity, and reduced aggregation.

[0073] The amino acid sequence of human IL-4RαECD is defined by SEQ ID NO: 15, the amino acid sequence of cynomolgus monkey IL-4RαECD is defined by SEQ ID NO: 16, the amino acid sequence of human IL-4 is defined by SEQ ID NO: 17, and the amino acid sequence of human IL-13 is defined by SEQ ID NO: 18.

[0074] The antibodies of the present invention can be synthesized and purified by well-known methods. Suitable host cells, such as Chinese hamster ovary cells (CHO), can be transiently or stably transfected with an expression system to secrete antibodies using a predetermined HC:LC vector ratio when two vectors are used, or using a single vector system encoding both the heavy and light chains. Clarified medium into which the antibodies are secreted can be purified using commonly used techniques.

[0075] Antibody engineering of IL-4Rα antibodies: The IL-4Rα antibody 5F3 IgG4PAA was engineered as a Fab in a mammalian cell expression vector using a high-throughput site-directed saturation mutagenesis protocol to find mutations that improve affinity and / or biophysical properties (e.g., thermal, chemical stability, or solubility, reduce aggregation or hydrophobicity). 5F3 IgG4PAA contains amino acid residue substitutions F234A and L235A in the IgG4 Fc region, which reduce binding to FcγRs, and amino acid substitution S228P, which stabilizes the hinge and prevents arm exchange.

[0076] Briefly, for engineering, all amino acids in the CDRs of both the VL and VH chains of 5F3 IgG4 PAA were mutated in individual mutagenesis reactions to generate a total of 18 variants (except cysteines), which were then reverted to the original amino acid residues (constituting an embedded wild-type control, WT) using a series of forward and reverse oligos arrayed in a 384-well microtiter plate. Site-directed mutagenesis reactions were performed according to established protocols, and digestion of the WT plasmid was achieved by incubation with DpnI restriction enzyme. The digested products were transformed into E. coli and incubated overnight at 37°C, after which DNA was isolated from the bulk transformants. DNA from each individual VL and VH mutagenesis reaction was mixed with the appropriate WT antibody variable region and expressed in CHO cells in 96-deep-well plates. Secreted antibodies were quantified and normalized to consistent titers before being screened for binding to IL4Rα in an ELISA format, with or without a heat challenge step. In addition to evaluating CDR variants, variants that convert atypical germline residues in the framework regions to more typical amino acids were also evaluated.

[0077] Hits were validated by ELISA titration, Octet, or Biacore 8 K, then guided by structure-based considerations, selected, combined, introduced into full-length antibody formats, and evaluated for affinity and biophysical properties using butyl-HIC, heparin, column interaction and size exclusion chromatography, differential scanning calorimetry, and serum protein binding by mass spectrometry.

[0078] Affinity binding analysis showed that the 5F3 IgG4PAA antibody has moderate affinity for human and cynomolgus monkey IL-4Rα, with K D is 10 -9 Mutagenesis of CDR amino acid residues in 5F3 IgG4PAA identified CDR substitutions: LCDR3 H91W, N92S, which resulted in a 10 M range of the resulting antibody. -11 The affinity for the M range was significantly improved.

[0079] Additionally, amino acid residue substitutions were identified that confer improved thermostability in heat challenge ELISA: VH: A23V, N92S, I31H; VL: G28D. Furthermore, amino acid residue substitutions: VH: A23V, I58V; VL: G28D were found to reduce self-association and hydrophobicity while maintaining affinity. Amino acid residue substitution: VH: I31H was found to reduce serum protein binding.

[0080] Furthermore, the 5F3 IgG4 PAA antibody contains an asparagine in HC framework 3 (N72) that deamidates under stressful conditions. Substitution of the N72 amino acid residue with the more germline Asp (N72D) eliminated the deamidation.

[0081] Mutagenesis analysis of 5F3 IgG4PAA identified seven amino acid residues for manipulation. These seven amino acid residues were substituted as follows: I31H, I58V, N72D in the VH region and H91W and N92S in the VL region were combined to generate the 8660 antibody variant; these five substitutions, plus A23V in the VH region and G28D in the VL region, created the 5559 antibody variant. Table 1 shows the CDR amino acid sequences of the exemplary antibodies.

[0082] Several versions of the 5559 and 8660 antibodies with different IgG backbones were generated, including those provided in Table 2. Manipulation of seven amino acid residues resulted in 5559 antibody variants with significantly improved affinity, as well as other biophysical properties such as thermal stability, reduced self-association, hydrophobicity, and / or serum protein binding, while maintaining affinity.

[0083] Antibody constant region engineering to improve viscosity: To improve viscosity and reduce potential electrostatic interactions between the antibody's Fab and constant domains, the 5559 IgG4 human IL-4Rα antibody heavy chain constant region was engineered by charge balancing. The CH1, CH2, and CH3 domains in the HC constant region of human IgG4 antibodies have lower isoelectric points (pI) due to uneven charge distribution compared to the human IgG1 HC constant region. Therefore, five important amino acid residues in the CH1, CH2, and CH3 domains that affect the viscosity of the 5559 IgG4 antibody were identified: 1) E137 (CH1 domain), 2) D203 (CH1 domain), 3) Q274 (CH2 domain), 4) Q355 (CH3 domain), and 5) E419 (CH3 domain). The analogous positions in the hIgG1 constant region for these five amino acids were found to be different and affect the overall pI of each domain.

[0084] To match the pI of the CH2 and CH3 domains of the IgG4 antibody to that of the IgG1 antibody and minimize the potential introduction of immunogenic peptides, three residues of the five identified positions in the IgG4 constant region were converted to the corresponding residues found in the IgG1 constant region. The amino acid residue substitutions included a positively charged lysine (Q274K) instead of a neutrally charged glutamine at position 274, a positively charged arginine (Q355R) instead of a neutrally charged glutamine at position 355, and a neutrally charged glutamine (E419Q) instead of a negatively charged glutamic acid at position 419. The resulting IgG4 Fc was designated "KRQ."

[0085] An IgG4 constant region containing substitutions at all five identified amino acid residues with those found in IgG1 (E137G, D203N, Q274K, Q355R, and E419Q) was also constructed and designated "GNKRQ."

[0086] The IgG4 KRQ and IgG4 GNKRQ antibodies also contained the S228P mutation, designated IgG4P, which stabilizes the hinge and prevents arm exchange. The wild-type IgG4 CH1 domain was used with a human kappa constant domain to complete the construct. The antibodies were synthesized, expressed, and purified essentially as described above.

[0087] Selection of IgG4P or IgG1A Scaffold: Human IgG1A and / or human IgG4P scaffolds were chosen for the exemplified 5559 antibody due to its unexpected binding properties to B cells and myeloid cells. As demonstrated in Table 5B and Figure 4C, the exemplified 5559 IgG4P and 5559 IgG1A IL-4Rα antibodies were found to have greater binding affinity to B cells when compared to the 5559 IgG1AAA effector-null antibody, thus indicating that the Fc portion of the 5559 antibody, which was not engineered to be effector-null, positively influenced B cell binding.

[0088] [Table 1]

[0089] [Table 2]

[0090] Example 2: Structural and functional epitopes of human IL-4Rα antibodies The structural epitopes of exemplary anti-IL-4Rα antibodies were determined by X-ray crystallography, and the functional epitopes of exemplary anti-IL-4Rα antibodies were determined by ELISA.

[0091] Example 2a. Structural epitope determination of 8660 Fab by X-ray crystallography The physical epitope of the 8660 anti-IL-4Rα antibody Fab on human IL-4Rα was determined by identifying the interaction interface between human IL-4Rα and the illustrated antibody. Briefly, to determine the structural epitope, human IL-4Rα ECD was co-crystallized with the 8660 Fab portion. The structure of 8660 Fab complexed with IL-4Rα was determined by generating a hexahistidine-tagged IgG1 variant of the heavy chain truncated after the CH1 domain and a "Crystal Kappa" version of the 8660 Fab light chain (see Lieu et al., "Rapid and Robust Antibody Fab Fragment Crystallization Utilizing Edge-to-edge Beta-sheet Packing," PLoS One, 15(9) (2020), incorporated herein by reference in its entirety). The 8660 variant was coexpressed with a hexahistidine-tagged version of human IL-4Rα ECD containing the C182L mutation, and the complex was then purified by immobilized metal affinity chromatography and screened using standard commercially available screens for crystallization. Crystals were obtained, and X-ray diffraction data were collected at an Advanced Photon Source. The diffraction data were reduced and solved by molecular replacement, and refined to obtain a 2.8 Å structure of the illustrated 8660 Fab and IL-4Rα ECD complex. From the resulting crystal structure, any IL-4Rα amino acid residue within 4.5 Å of an atom of the cocrystallized 8660 Fab was counted as part of the epitope (using PyMOL visualization software [Schrodinger®]).

[0092] PyMOL analysis demonstrated that IL-4Rα amino acid residues (with respect to SEQ ID NO: 15) within 4.5 Å of the 8660 Fab in the crystal structure complex comprise the structural epitope of the exemplified antibody. Specifically, the analysis determined that the structural epitope includes the following amino acid residues: Asp at position 12, Met at position 14, Ser at position 15, Ile at position 16, Tyr at position 37, Leu at position 39, Phe at position 41, Leu at position 43, Glu at position 45, His at position 47, Thr at position 48, Cys at position 49, Ile at position 50, His at position 62, Leu at position 64, Met at position 65, Asp at position 66, Asp at position 67, Val at position 69, Asp at position 72, Arg at position 99, Pro at position 121, Pro at position 123, Pro at position 124, and Asp at position 125. The analysis further determined that the structural epitope spans domains 1 and 2 of the N-terminal fibronectin type III domain of IL-4Rα. Furthermore, analysis determined that the following amino acid residues of the structural epitope are located in domain 2 of the N-terminal fibronectin type III domain of IL-4Rα: R99, P121, P123, P124, and D125.

[0093] Additionally, an overlay of the illustrated crystal structures of 8660 Fab and dupilumab Fab with a crystalline kappa design complexed with human IL-4Rα (pdb accession code 6WGL) showed that 8660 Fab binds to a novel epitope on IL-4Rα when compared to dupilumab (Figure 1).

[0094] Furthermore, alignment of the exemplified IL-4Rα 8660 Fab:IL-4Rα complex crystal structure with the published complexes of IL-4 and IL-13 and their respective receptors (pdb accession codes 3BPN and 3BPO) on the IL-4Rα component in each structure (using PyMOL visualization software) shows that the exemplified 8660 Fab antibody epitope overlaps with both the IL-4 and IL-13 binding sites for IL-4Rα, thus indicating that when the Fab variant portion of the exemplified antibody binds to IL-4Rα, binding of the exemplified antibody to IL-4Rα physically blocks the binding of IL-4 and IL-13 cytokines to IL-4Rα.

[0095] Example 2b. Functional epitope determination of 5F3 IgG4PAA The functional epitope of the exemplary human IL-4Rα antibody 5F3 IgG4PAA was determined by ELISA. Briefly, 30 surface amino acid residue substitutions were individually introduced into a hexahistidine-tagged human IL-4Rα extracellular domain (ECD) as follows: K2D, E6R, K22D, P26R, T31R, F41A, L42G, L43G, E45R, G56R, D66R, A71R, Q82G, K87D, E94R, H107A, D108R, P124R, D125R, D143R, R148D, L155R, R160D, S164R, S168R, Q181R, P192R, K195D, or H197G. Each mutein with a single amino acid residue substitution as described above was transiently expressed in CHO cells and purified using standard immobilized metal affinity chromatography techniques. ELISA plates were coated with 1 μg / mL goat anti-human kappa antibody (Southern Biotech, catalog no. 2060-01) in PBS overnight at 4°C, then washed three times with PBST and blocked with PBS-casein for 30 minutes at room temperature. The plates were then washed three times with PBST, and the exemplified human IL-4Rα antibody 5F3 IgG4PAA was added to the wells at a final concentration of 1 μg / mL in PBS-casein and incubated for 1 hour. The plates were then washed three times with PBST, and the IL-4Rα muteins were serially diluted three-fold starting from 1 μg / mL in PBS-casein, added to the plate at 50 μL / well, and incubated for 1 hour at room temperature. The plate was washed three times with PBST, and a 5000-fold dilution of anti-histidine tag antibody HRP conjugate (R&D Systems, catalog no. MAB050H) in PBS-casein was added and incubated for 1 hour at room temperature. The plate was washed three times, and TMB substrate (Pierce, catalog no. 34021) was added according to the manufacturer's instructions. The reaction was quenched with H2SO4, and absorbance was read at 450 nm on an ELISA plate reader. The functional epitope of the antibody was determined as the mutated amino acid residue corresponding to wells that showed either no binding signal or a significantly reduced binding signal compared to the control antibody.

[0096] The results shown in Table 3 indicate that the functional epitope of the exemplified 5F3 IgG4PAA antibody comprises amino acid residues D66 and D125. Among the amino acid residues identified in the structural epitope, amino acid residue substitutions of D66R and D125R on IL-4Rα each showed a significantly negative effect on the binding of the exemplified 5F3 IgG4PAA to the mutant IL-4Rα. Specifically, substitution of amino acid residue D66 of IL-4Rα with arginine reduced the binding of 5F3 IgG4PAA to the mutant IL-4Rα compared to the binding of the control (0.04 OD, respectively). 450 and 0.14OD 450 Furthermore, substitution of amino acid residue D125, located near amino acid residue D66 in the crystal structure of IL-4Rα, with arginine (see Figure 2) also resulted in a 0.59 OD 450 The remaining amino acid substitutions were either within the range of positive binding or outside the determined structural epitope.

[0097] [Table 3]

[0098] Example 2c: Conformational epitope of human IL-4Rα5559 antibody Structural Epitope Determination of 5559 Fab by X-ray Crystallography. The physical epitope of the 5559 antibody Fab on human IL-4Rα was determined essentially as described above. Crystals were obtained, and X-ray diffraction data were collected at an Advanced Photon Source. The diffraction data were reduced and solved by molecular replacement, and refined to obtain a 2.49 Å structure of the illustrated 5559 Fab and IL-4Rα ECD complex. From the resulting crystal structure, any IL-4Rα amino acid residue within 4.5 Å of an atom of the co-crystallized 5559 Fab was counted as part of the epitope (using Molecular Operating Environment (MOE) visualization, modeling, and simulation software [Chemical Computing Group], Coot (General Public License), and PyMOL visualization software [Schrodinger®]).

[0099] MOE, Coot, and PyMOL analyses demonstrated that IL-4Rα amino acid residues (with respect to SEQ ID NO: 15) within 4.5 Å of the 5559 Fab in the crystal structure complex comprise a structural epitope. Specifically, the analyses determined that the structural epitope comprises the following amino acid residues: The following residues were identified: Asp at position 12, Met at position 14, Ser at position 15, Ile at position 16, Leu at position 39, Phe at position 41, Leu at position 42, Thr at position 48, Cys at position 49, Ile at position 50, Glu at position 52, His at position 62, Leu at position 64, Met at position 65, Asp at position 66, Asp at position 67, Val at position 68, Val at position 69, Asp at position 72, Arg at position 99, Pro at position 121, Pro at position 123, Pro at position 124, Asp at position 125, and Pro at position 192. Asp at position 66 was well-matched with the heavy chain of 5559 Fab, interacting between 2.6 and 2.9 Å. Asp at position 67 had a longer-range interaction between 3.1 and 3.5 Å and showed flexibility in its binding position, as evidenced by the observation of excess density around its side chain. Analysis determined that the structural epitope spans domains 1 and 2 of the N-terminal fibronectin type III domain of IL-4Rα. Furthermore, analysis determined that the following amino acid residues of the structural epitope are located in domain 2 of the N-terminal fibronectin type III domain of IL-4Rα: R99, P121, P123, P124, D125, and P192.

[0100] An overlay of the crystal structures of the illustrated 5559 Fab and dupilimab Fab with the crystal kappa design complexed with human IL-4Rα (pdb accession code 6WGL) showed that the 5559 Fab bound to a novel epitope on IL-4Rα when compared to dupilimab (Figure 13).

[0101] Alignment of the exemplified IL-4Rα 5559 Fab: Alignment (using PyMOL visualization software) of the IL-4Rα complex crystal structure with published complexes of IL-4 and IL-13 on IL-4Rα and their respective receptors (pdb accession codes 3BPN and 3BPO) in each structure showed that the exemplified 5559 Fab antibody epitope overlaps with both the IL-4 and IL-13 binding sites on IL-4Rα, indicating that binding of the exemplified 5559 antibody would physically block the binding of IL-4 and IL-13 cytokines to IL-4Rα.

[0102] Example 2d. Determination of the functional epitope of the human IL-4Rα5559 antibody The functional epitope of the exemplary human IL-4Rα antibody 5559 IgG4P KRQ was determined by ELISA. Briefly, 30 surface amino acid residue substitutions were individually introduced into a hexahistidine-tagged human IL-4Rα extracellular domain (ECD) as follows: K2D, E6R, K22D, P26R, T31R, F41A, L42G, L43G, E45R, E52R, G56R, D66R, A71R, Q82G, K87D, E94R, H107A, D108R, P124R, D125R, D143R, R148D, L155R, R160D, S164R, S168R, Q181R, P192R, K195D, or H197G. Each mutein with a single amino acid residue substitution as described above was transiently expressed in CHO cells and purified using standard immobilized metal affinity chromatography techniques. ELISA plates were coated with 1 μg / mL goat anti-human IgG Fc antibody (Jackson ImmunoResearch Laboratories, catalog number 109-005-098) in PBS overnight at 4°C, then washed three times with PBST and blocked with PBS-casein for 1 hour at room temperature. The plates were then washed three times with PBST, and the illustrative human IL-4Rα antibody 5559 IgG4P KRQ was added to the wells at a final concentration of 1 μg / mL in PBS-casein and incubated for 1 hour at room temperature. The plates were then washed three times with PBST, and the IL-4Rα muteins were serially diluted 5-fold in triplicate from 1 μg / mL in PBS-casein, added to the plate at 50 μL / well, and incubated for 1 hour at room temperature. The plate was washed three times with PBST, and a 1:1000 dilution of anti-histidine tag antibody HRP conjugate (R&D Systems, catalog no. MAB050H) in PBS-casein was added and incubated for 45 minutes at room temperature. The plate was washed three times, and TMB substrate (Pierce, catalog no. 34028) was added according to the manufacturer's instructions. The reaction was quenched with H2SO4, and absorbance was read at 450 nm on an ELISA plate reader. The functional epitope of the antibody was determined as the mutated amino acid residue corresponding to wells that showed either no binding signal or a significantly reduced binding signal compared to the wild-type control.

[0103] The results shown in Table 4 indicate that the functional epitope of the exemplary 5559 IgG4P KRQ antibody includes amino acid residue D66. Substitution of amino acid residue D66 to D66R on IL-4Rα reduced binding of 5559 IgG4P KRQ to D66R IL-4Rα compared to binding of the negative control (0.047 OD and 0.057 OD , respectively). 450 and 0.063OD 450 ). Amino acid residue D66 is located near the structural epitope residues D67 and D125 in the crystal structure of IL-4Rα (Figure 14). The remaining amino acid substitutions were either within the range of positive binding or outside the determined structural epitope.

[0104] [Table 4]

[0105] Example 3. Binding affinity and functional activity of human IL-4Rα antibodies Binding affinity: The binding affinity of the exemplified anti-IL-4Rα antibodies for human and cynomolgus IL-4Rα was measured using a competitive Meso Scale Discovery (MSD) ELISA binding assay. A fixed concentration of each antibody was mixed with a 3-fold dilution series of IL-4Rα to obtain final concentrations of 10 pM for each 5559 antibody and 100 pM for 5F3. Starting dilutions were performed at 10 nM IL-4Rα for 5559 antibody and 200 nM IL-4Rα for 5F3, and the mixtures were incubated at 37°C for 4 days. A 96-well multi-array plate (Meso Scale Diagnostics, catalog number L15XA-3) was coated overnight at 4°C with 0.5 μg / mL hexahistidine-tagged human or cynomolgus IL-4Rα ECD in phosphate-buffered saline (PBS). After coating, the plates were washed 10 times with 200 μL of PBST (PBS with 0.05% Tween® 20) and blocked with 150 μL of PBS-casein blocking buffer (Pierce, catalog number 37528) for 1 hour at 37°C. The plates were then washed 10 times as above, and 50 μL of the pre-incubated antibody:IL-4Rα dilution series was transferred to the wells and incubated for 150 seconds at 37°C with shaking at 300 rpm. The plates were washed 10 times with PBST, and 50 μL of 1 μg / mL anti-human antibody sulfo-tag20 (Meso Scale Diagnostics, catalog number #R32AJ-1) was added, and the plates were incubated for 30 minutes at 37°C with shaking at 300 rpm. Plates were washed 10 times with PBST, and 150 μL / well of 1× Read Buffer T was added to wells. 15 minutes after buffer addition, the plates were analyzed on a SECTOR® Imager 6000 (Meso Scale Diagnostics). Apparent KDs were determined by fitting a sigmoidal curve to the electrochemiluminescence (ECL) response versus log (soluble IL-4Rα concentration) using GraphPad Prism 9. Data were graphed using normalized ECL values.

[0106] The results, shown in Table 4 and Figures 3A and 3B, demonstrate that the exemplified 5559 anti-IL-4Rα antibody variants had significantly increased binding affinity to both human and cynomolgus IL4-Rα when compared to 5F3 IgG4PAA. Specifically, the binding affinity of each 5559 antibody variant to human IL4-Rα was 5559 IgG1A 124C / 378C (45.08 pM), 5559 IgG4P KRQ 124C / 378C (20.82 pM), and 5559 IgG4P 124C / 378C (24.63 pM), when compared to 5F3 IgG4PAA (1993 pM).

[0107] [Table 5]

[0108] Binding to B and T cells: Binding of the exemplary human IL-4Rα antibodies to B and T cells was tested in a fluorescence-activated cell sorting (FACS) assay. Human PBMCs were isolated from human blood samples by standard Ficoll-Paque™ plus (GE HEALTHCARE) density gradient centrifugation. Freshly isolated PBMCs were cultured at 2 x 10 6The cells were resuspended at 100 μL / mL and incubated at room temperature for 15 minutes, then seeded at 100 μL / well into a round-bottom 96-well plate (COSTAR®) and washed with FACS buffer (PBS containing 2% fetal bovine serum from Corning®). The exemplified human IL-4Rα antibody and the respective control IgG antibody conjugated to Alexa Fluor® 647 were added to wells at 66.67 nM and diluted 4-fold in duplicate according to the manufacturer's protocol (Thermo Fisher Scientific). An equal volume of a 2× antibody cocktail containing human TruStain FcX™, FITC anti-human CD3 antibody, Alexa Fluor® 700 anti-human CD4 antibody (all from Biolegend®), and CD20 monoclonal antibody (2H7), PerCP-Cyanine 5.5 (Thermo Fisher Scientific), was then added to the wells. Cells were incubated at 4°C for 30 minutes, washed twice with FACS buffer, and resuspended in a final volume of 100 μL of FACS buffer. Viability dye, Sytox™ blue (Thermo Fisher Scientific), was added, and samples were analyzed via a flow cytometer (LSRFortessa™ X-20; BD BIOSCIENCES). Data analysis was performed using FlowJo software, and statistical analysis was performed using GraphPad Prism 9. Data represent the mean ± SEM of the percentage of IL-4Rα-expressing cells from CD20 B cell and CD4+ T cell populations from six donors. Curves were generated by fitting sigmoidal curves of log(Ab concentration) vs. percent of positive IL-4Rα-expressing cells from individual cell populations.

[0109] The results, shown in Table 5A and Figures 4A and 4B, demonstrate that the exemplary IL-4Rα antibodies 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C inhibited the IL-4Rα activity of human PBMC-isolated B cells (EC of 0.14 nM and 0.15 nM, respectively). 50 ) and CD4 T cells (EC of 28.7 nM and 26.3 nM, respectively) 50) with an affinity comparable to that of the IL-4Rα5559 antibody. Furthermore, the results showed that the KRQ and 124C / 378C amino acid residue substitutions did not affect the binding of the exemplified anti-IL-4Rα5559 antibody to B cells or T cells.

[0110] Furthermore, as demonstrated in Table 5B and Figure 4C, the 5559 IgG1AAA 124C / 378C effector null antibody exhibited a similar activity to the 5559 IgG1A 124C / 378C antibody (EC 50 ) showed an unexpectedly reduced affinity for B cells (EC of 1.07 nM) compared to 50 ), indicating that the Fc portion of an antibody can affect the binding of an exemplary IL-4Rα antibody to B cells.

[0111] [Table 6]

[0112] [Table 7]

[0113] Cell-based IL-4 and IL-13 cytokine blocking activity: The antagonist activity of the exemplified anti-IL-4Rα antibodies against IL-4 and IL-13 was determined using the HEK-Blue IL-4R and IL-13R expressing cell line (InvivoGen) by measuring secreted embryonic alkaline phosphatase (SEAP) activity. HEK-Blue cells were plated at 5 x 10 in 50 μL of growth medium in polylysine-coated plates. 4Cells / well were seeded overnight. Exemplary IL-4Rα antibodies (5F3 IgG4PAA, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ 124C / 378C, 5559 IgG1A 124C / 378C, and 8660 IgG4P 124C / 378C) were prepared in 4-fold dilutions starting at 20 μg / mL in growth medium in a Greiner 96-well low protein binding plate. The dilution series was mixed with an equal volume of either recombinant human IL-4 or IL-13 (Eli Lilly) in growth medium. 50 μL of the mixture was then added to the plate with HEK-Blue cells to achieve a final concentration of 100 pg / mL human IL-4 or 10 ng / mL human IL-13, and the plate was then incubated overnight at 37°C in a tissue culture incubator. 20 μL of supernatant from the overnight incubation plate is transferred to a 96-well tissue culture-treated plate, 180 μL of QUANTI-Blue™ (InvivoGen) is added per well, and the mixture is incubated at 37°C for 45 minutes. Secreted embryonic alkaline phosphatase (SEAP) activity was measured at 650 nm in a SpectraMax microplate reader (Molecular Devices). Results were reported as optical density (OD) at 650 nm, and statistical analysis was performed using GraphPad Prism 9. IC 50 and curves were generated by fitting a sigmoidal curve of log(Ab concentration) vs. OD at 650 nm for each example antibody.

[0114] The results, shown in Table 6 and Figures 5A and 5B, demonstrate that the exemplary IL-4Rα antibodies 5559 IgG1A 124C / 378C, 5559 IgG4P KRQ 124C / 378C, and 8660 IgG4P 124C / 378C inhibited both IL-4 (Figure 5A) and IL-13 (Figure 5B)-induced SEAP activity in a dose-dependent manner. Specifically, as shown in Table 6 and Figure 5A, inhibition of IL-4-induced SEAP activity for the 5559 IgG1A 124C / 378C, 5559 IgG4P KRQ 124C / 378C, and 8660 IgG4P 124C / 378C antibodies was with IC s of 0.07, 0.08, and 0.03 nM, respectively. 50 Furthermore, as shown in Table 6 and Figure 5B, inhibition of IL-13-induced SEAP activity for the 5559 IgG1A 124C / 378C, 5559 IgG4P KRQ 124C / 378C, and 8660 IgG4P 124C / 378C antibodies yielded comparable IC values ​​of 0.51, 0.67, and 0.24 nM, respectively. 50 brought value.

[0115] [Table 8]

[0116] Inhibition of IL-4- and IL-13-induced pSTAT6 phosphorylation in human PBMCs: Inhibition of IL-4- and IL-13-mediated IL-4R pSTAT6 phosphorylation by the exemplary anti-IL-4Rα antibodies was evaluated in primary B cells and / or primary T cells. Human PBMCs were isolated from human blood samples by standard Ficoll-Paque™ plus (GE HEALTHCARE) density gradient centrifugation. Isolated cells were resuspended at 100-300 million cells in 100 mL of complete medium (10% FBS, 1% penicillin-streptomycin solution from Corning®, and 1% GlutaMAX™ and 0.1% β-mercaptoethanol, RPMI-1640 from Gibco™) in a T175 flask (FALCON) and stimulated overnight with 2 μg / mL PHA (SIGMA), 0.5 μg / mL LPS (SIGMA), and 100 ng / mL recombinant human IL-6. Cells were washed with fresh medium and plated at 5 x 10 cells in a 96-well round-bottom plate (Corning®) in 100 μL complete medium containing 10 μg / mL of an exemplary antibody diluted in 4-fold dilutions and an 11-point titration. 4 ~2×10 5Cells were seeded at 1000 x g / well. Cells were incubated with antibody for 30 minutes at room temperature and then stimulated with 120 ng / mL (6x concentration) human recombinant IL-4 or human recombinant IL-13 (R&D SYSTEMS) in 20 μL complete medium for 12 minutes at room temperature. Stimulation was stopped by adding 120 μL of 1x Lyse / Fix Buffer (BD BIOSCIENCES) for 5 minutes. The plate was then centrifuged at 2000 rpm for 2 minutes and the supernatant was aspirated. The cell pellet was resuspended in 100 μL of ice-cold methanol (SIGMA), placed on ice for 20 minutes, and washed with DPBS containing 2% FBS (Corning®). Cells were resuspended in 50 μL of an antibody cocktail against the following proteins: CD4, CD33, CD8, and CD3 (Thermo Fisher Scientific), phosphorylated STAT6 (Biolegend®), and CD20 (BD BIOSCIENCES), incubated at room temperature for 30 minutes, and then washed with DPBS containing 2% FBS. Cell samples were analyzed using a flow cytometer. Analysis was performed using FlowJo software, and statistical analysis was performed using GraphPad Prism 9. Curves were generated by fitting a sigmoidal curve of log (Ab concentration) versus percent inhibition of phosphorylated STAT6 from individual cell populations derived from two donors.

[0117] The results, shown in Table 7 and Figures 6A and 6B, demonstrate that the exemplary IL-4Rα antibodies dose-dependently inhibited IL-4-induced pSTAT6 phosphorylation in both CD4+ T cells (Figure 6A) and B cells (Figure 6B). Specifically, the IC for inhibition of IL-4-induced pSTAT6 phosphorylation by the exemplary 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C antibodies was 0.01. 50 Values ​​were 0.07 μg / mL in CD4 T cells and 0.05 μg / mL in B cells for both antibodies.

[0118] Furthermore, the results shown in Table 8 and Figure 6C indicate that the exemplary IL-4Rα antibodies also dose-dependently inhibited IL-13-induced pSTAT6 phosphorylation in B cells (Figure 6C). Specifically, the IC values ​​for the 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C antibodies for inhibition of IL-13-induced pSTAT6 phosphorylation in B cells were 0.01 and 0.02, respectively. 50 The value was 0.06 μg / mL for both antibodies.

[0119] [Table 9]

[0120] [Table 10]

[0121] Inhibition of IL-4-induced B cell proliferation: Inhibition of B cell proliferation by the exemplary human IL-4Rα antibodies was evaluated in primary B cells isolated from human PBMCs. Human PBMCs were isolated from human blood samples by standard Ficoll-Paque™ plus (GE HEALTHCARE) density gradient centrifugation, and primary B cells were isolated from the PBMC suspension by negative selection using the EasySep™ Human Naive B Cell Enrichment Kit (STEMCELL™ technology) according to the manufacturer's protocol. 1 x 10 isolated human primary B cells were cultured at 1 x 10 6Cells were resuspended at 37°C / mL and seeded into polystyrene 96-well U-bottom plates in complete medium (RPMI-1640 containing 10% fetal bovine serum, 1x MEM-non-essential amino acids, 1 mM sodium pyruvate, 1x penicillin-streptomycin solution (all from Corning®), and 1x GlutaMAX™ (Gibco™), 0.1% β-mercaptoethanol (LIFE TECHNOLOGIES). Cells were pretreated for 0.5-1 h with anti-IL-4Rα antibody or isotype control at 66.67 nM and a 10-point titration in four-fold dilutions. Cells were stimulated with human CD40 / TNFRSF5 antibody (200 ng / mL; R&D SYSTEMS) and IL-4 recombinant human protein (5 ng / mL; R&D SYSTEMS) for 2 days at 37°C and 5% CO2. Cells were then incubated for 2 h with 100 μL of IL-4Rα antibody or isotype control at 66.67 nM and a 10-point titration in four-fold dilutions. 3 H]-thymidine (1 μCi thymidine / well; PerkinElmer®) for 18 hours at 37°C, and 3 The level of [H]-thymidine incorporation was measured by a microplate counter (MicroBeta2; PerkinElmer®) and expressed as cell counts per minute (CCPM). Statistical analysis was performed using GraphPad Prism 9, and curves were generated by sigmoidal curve fitting of log(Ab concentration) vs. CCPM.

[0122] The results shown in Table 9 and Figure 7 demonstrate that the exemplary IL-4Rα antibodies inhibited IL-4-induced B cell proliferation in a dose-dependent manner. Specifically, the IC for inhibition of IL-4-induced B cell proliferation by the 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C antibodies was 0.01. 50 were 0.95 nM and 1.32 nM, respectively.

[0123] [Table 11]

[0124] Inhibition of IL-4 and IL-13-induced CD23 expression in myeloid cells: Inhibition of IL-4 and IL-13-induced CD23 expression by the exemplary human IL4Rα antibodies was evaluated in myeloid cells. Human PBMCs were isolated from human blood samples by standard Ficoll-Paque™ plus (GE HEALTHCARE) density gradient centrifugation. Cells were plated at 2 x 10 in a 96-well flat-bottom plate. 5 Cells were plated at 1000 x g / well. 50 μL of 3x serially diluted antibody was added to the wells and incubated at 37°C, 5% CO2 for 30 minutes. Then, 50 μL of a triple stimulation of either recombinant human IL-4 or IL-13 (R&D SYSTEMS) in complete medium was added to the wells to a final concentration of 10 ng / mL. The plates were incubated at 37°C with 5% CO2 for 48 hours, after which the cells were washed and resuspended in FACS buffer containing Human TruStain FcX™, Brilliant Violet 785™ anti-human CD33 antibody, FITC anti-human CD3 antibody (Biolegend®), CD20 monoclonal antibody (2H7) PerCP-Cyanine 5.5, and CD23 monoclonal antibody (EBVCS 2), APC (Thermo Fisher Scientific). Cells were incubated for 30 minutes at 4°C, washed twice with FACS buffer, and resuspended in a final volume of 100 μL of FACS buffer. Viability dye, Sytox™ blue (THERMO FISHER SCIENTIFIC), was added to the wells, and samples were analyzed using a flow cytometer (LSRFortessa™ X-20; BDBIOSCIENCES). Data analysis was performed using FlowJo software. Myeloid cells were identified as Sytox™ blue-, CD3-, and CD20-negative, CD33-positive cells. Data are presented as sigmoidal curve fits of percent inhibition versus log(Ab concentration) for two donors, and statistical analysis was performed using GraphPad Prism 9.

[0125] The results shown in Table 10 and Figures 8A and 8B demonstrate that the exemplary IL-4Rα antibodies inhibited both IL-4 (Figure 8A) and IL-13 (Figure 8B)-induced CD23 expression on bone marrow cells. Specifically, the IC for inhibition of IL-4-induced CD23 expression by the 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C antibodies was 0.01. 50 The IC values ​​for inhibition of IL-13-induced CD23 expression by the 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C antibodies were 4.44 nM and 18.25 nM, respectively. 50 The values ​​were 1.28 nM and 5.37 nM, respectively.

[0126] [Table 12]

[0127] Example 4. Effector function activity of human IL-4Rα antibody Human Fcγ Receptor Binding. The binding affinity of the exemplified anti-IL-4Rα antibodies to human Fcγ receptors was assessed by surface plasmon resonance (SPR) analysis. Series S CM5 chips (Cytiva P / N BR100530) were prepared using the manufacturer's EDC / NHS amine coupling method (Cytiva P / N BR100050). Briefly, the surfaces of all four flow cells (FCs) were activated by injecting a 1:1 mixture of EDC / NHS at 10 μL / min for 7 minutes. Protein A (Calbiochem P / N 539202) was diluted to 100 μg / mL in 10 mM acetate buffer, pH 4.5, and immobilized to approximately 4000 RU on all four FCs by injecting at a flow rate of 10 μL / min for 7 minutes. Unreacted sites were blocked by injecting ethanolamine at 10 μL / min for 7 minutes. Non-covalently associated proteins were removed using 2 x 10 µL injections of glycine at pH 1.5. The running buffer was 1x HBS-EP+ (TEKNOVA, P / N H8022). FcγR extracellular domain (ECD)-FcγRI (CD64), FcγRIIA_131R, and FcγRIIA_131H (CD32a), FcγRIIIA_158V, FcγRIIIA_158F (CD16a), and FcγRIIb (CD32b) were produced from stable CHO cell expression and purified using IgG Sepharose and size-exclusion chromatography. For FcγRI binding, antibodies were diluted to 2.5 µg / mL in running buffer, and approximately 150 RU of each antibody was captured in FCs 2–4 (RU capture). FC1 was the reference FC; therefore, no antibody was captured in FC1. FcγRI ECD was diluted to 200 nM in running buffer and then serially diluted two-fold to 0.78 nM in running buffer. Duplicate injections of each concentration were injected into all FCs at 40 μL / min for 120 seconds, followed by a 1200-second dissociation phase. Regeneration was performed by injecting 15 μL of 10 mM glycine, pH 1.5, into all FCs at 30 μL / min. Reference-subtracted data were collected for FC2, FC1, FC3-FC1, and FC4-FC1, and measurements were taken at 25°C.Affinity (KD) was calculated using either steady-state equilibrium analysis or a "1:1 (Langmuir) binding" model for BIA analysis using Scrubber2 Biacore software. For FcγRIIa, FcγRIIb, and FcγRIIIa binding, antibodies were diluted to 5 μg / mL in running buffer, and approximately 500 RU of each antibody was captured in FCs 2–4. FC1 served as the reference FC. Fcγ receptor ECDs were diluted to 10 μM in running buffer and then serially diluted 2-fold in running buffer to 39 nM. Duplicate injections of each concentration were injected into all FCs at 40 μL / min for 60 s, followed by a 120-s dissociation phase. Regeneration was performed by injecting 15 μL of 10 mM glycine, pH 1.5, into all FCs at 30 μL / min. Reference subtracted data were collected as FC2 FC1, FC3-FC1, and FC4-FC1, and measurements were obtained at 25°C. Affinity (KD) was calculated using steady-state equilibrium analysis with Scrubber2 Biacore® evaluation software. Each receptor was assayed at least twice.

[0128] The results shown in Table 11 show the binding affinities (K D ) is a summary of

[0129] [Table 13]

[0130] C1q Binding. Binding of the exemplified anti-IL-4Rα antibodies to human C1q was assessed by ELISA. 96-well microplates were coated with 100 μL / well of each exemplified antibody diluted in DPBS (Dulbecco's HyClone) at 10 μg / mL to 0.19 μg / mL and incubated overnight at 4°C. The coating reagent was removed, and the plates were blocked with 200 μL / well of casein blocking buffer (Thermo) and incubated for 2 hours at room temperature (RT). The plates were washed three times with wash buffer (1x TBE containing 0.05% Tween 20), and 100 μL / well of 10 μg / mL human C1q (MS Biomedical) diluted in casein blocking reagent was added and incubated for 3 hours at room temperature. Humanized IgG1 and humanized IgG4P isotype control antibodies were used as positive and negative controls, respectively. The plate was then washed three times with wash buffer, followed by the addition of 100 μL / well of a 1:800 dilution of sheep anti-human C1q-HRP (Abcam #ab46191) in casein blocker and incubation for 1 hour at room temperature. The plate was then washed six times with wash buffer, and 100 μL / well of TMB substrate (Pierce) was added to each well and incubated for 7 minutes. The reaction was stopped by the addition of 100 μL / well of 1N HCl. Optical density was immediately measured at 450 nm using a colorimetric microplate reader. Data were analyzed using SoftMax Pro 7.1 data acquisition and analysis software.

[0131] The results shown in Figure 9 demonstrate that the exemplified antibodies 5559 IgG1A 124C / 378C and 5559 IgG4P KRQ 124C / 378C did not bind to complement component C1q.

[0132] Antibody-Dependent Cellular Cytotoxicity (ADCC): In vitro ADCC assays of the exemplified antibodies were evaluated in either reporter gene-based ADCC assays or primary human NK and Th2 cell-based ADCC assays.

[0133] For the reporter gene-based ADCC assay, Daudi cells (ATCC, #CCL-213) expressing human IL-4Rα and human CD20 were used as the target cell line, and Jurkat cells expressing functional FcγRIIIa(V158)-NFAT-Luc (Eli Lilly and Company) were used as the effector cell line. All test antibodies and cells were diluted in assay medium containing RPMI-1640 (phenol red-free) with 0.1 mM non-essential amino acids (NEAA), 1 mM sodium pyruvate, 2 mM L-glutamine, 500 U / mL penicillin-streptomycin, and 0.1% w / v BSA. Test antibodies were initially diluted to a 3x concentration of 3.3 μg / mL and then serially diluted 7x at a 1:4 ratio. 50 μL / well of each antibody was aliquoted in duplicate into a white, opaque-bottom 96-well plate (Costar, #3917). CD20 antibody was used as a positive control. Daudi target cells were then cultured at 5 × 10 in 50 μL aliquots. 4 100,000 cells / well were added to the plate and incubated at 37°C for 1 hour. Jurkat V158 cells were then added to the wells at 150,000 cells / well in 50 μL aliquots and incubated at 37°C for 4 hours, followed by the addition of 100 μL / well of One-Glo luciferase substrate (Promega, #E8130). The plate contents were mixed using a plate shaker at low speed, incubated at room temperature for 5 minutes, and the luminescence signal was read on a BioTek microplate reader (BioTek Instruments) using an integration of 0.2 cps. Data were analyzed using GraphPad Prism 9, and the relative luminescence units (RLU) for each antibody concentration were plotted in a scatter format of antibody concentration vs. RLU. Results were representative of two independent experiments.

[0134] For primary human NK and Th2 cell-based ADCC assays, human Th2 cells cocultured with primary human B cells and primary human NK cells were used. Human primary B cells, NK cells, and naive CD4 T cells were isolated from freshly purified human PBMCs by immunomagnetic negative selection according to the manufacturer's protocol (Human B cell Enrichment Kit, Stemcell Technologies #19054; Human NK Cell Isolation Kit, Stemcell Technologies #17955; Human Naive CD4 T Cell Isolation Kit II, Stemcell Technologies #17555). Human Th2 cells were differentiated in vitro by culturing purified naive CD4 T cells with anti-human CD3 (BioXCell #BE0001-2), anti-human CD28 (BioLegend #302934), anti-human IFNγ (R&D Systems #MAB285-500), recombinant human IL-2 (R&D Systems #202-IL-050 / CF), and recombinant human IL-4 (R&D Systems #6507-IL-100 / CF) for 14 days. Cell purity was assessed using flow cytometry staining on a BD LSRFortessa Cell Analyzer. NK cells were differentiated by CD56 + (anti-human CD56-PE / Dazzle-594, BioLegend #318348) and FcγRIII + (anti-human CD16-SuperBright-702, Fisher Scientific #67-0168-42), and B cells were identified as CD19 + (anti-human CD19-PE-Cy5, Fisher Scientific #15-0199-42) and IL-4Rα + (5559-Alexa Fluor-647, Lilly), and Th2 cells were identified as CD4 + (anti-human CD4-eFluor-450, Fisher Scientific #48-0047-42), GATA3 + (Anti-human GATA3-PerCP / Cyanine5.5, BioLegend #653812) and IL-4Rα +It was confirmed that 5×10 4 B cells or Th2 cells / well were treated with 30 μg / mL or 5 μg / mL of 5559 IgG1A 124C / 378C antibody, respectively, and 250,000 NK cells and 37 o Cells were co-cultured at 4°C for 24 hours. Positive control wells were treated with anti-human CD52 antibody (Eli Lilly and Company). ADCC was measured using the CytoTox-Glo Cytotoxicity Assay (Promega #G9292) according to the manufacturer's protocol. Relative luminescence was detected using a Biotek Cytation 5 Imaging Multi-Mode Reader. Data represent three technical replicates per donor. Statistical analysis was performed using GraphPad Prism 9. Data represent the mean + SD of relative luminescence units. Treatment differences were assessed using one-way ANOVA for each cell type individually, and group comparisons to the No Ab group were assessed using Tukey's test for multiple comparisons, with a significance level of 0.05.

[0135] The results of the reporter gene-based assay, as demonstrated in Figure 10A, show that the exemplified IL-4Rα antibodies 5559 IgG1A 124C / 378C, 5559 IgG4P KRQ 124C / 378C, and 5559 IgG4P 124C / 378C significantly lacked or had no ADCC activity when compared to the positive control.

[0136] Furthermore, as shown in FIG. 10B, the results of primary human NK and Th2 cell-based ADCC assays indicate that the exemplary IL-4Rα antibody 5559 IgG1A 124C / 378C did not induce ADCC activity in primary human B cells and human Th2 cells when compared to the positive control, and showed significant ADCC activity in both primary human B cells (p<0.0001) and Th2 cells (p=0.032) when compared to the negative control group.

[0137] Complement-dependent cytotoxicity (CDC): In vitro CDC assays of the exemplified antibodies were performed using Daudi cells (ATCC, #CCL-213). All test antibodies, complement, and cells were diluted in assay medium consisting of RPMI-1640 (phenol red-free) containing 0.1 mM non-essential amino acids (NEAA), 1 mM sodium pyruvate, 2 mM L-glutamine, 500 U / mL penicillin-streptomycin, and 0.1% w / v BSA. Test antibodies were initially diluted to a 3x concentration of 100 μg / mL, then serially diluted 7x at a 1:4 ratio. 50 μL / well of each antibody (including a CD20 positive control antibody) was aliquoted in duplicate into white, opaque-bottom 96-well plates (Costar, #3917). Daudi target cells were plated in 5x10 wells. 4 Cells / well were added at 50 μL / well and incubated for 1 hour at 37°C. Next, human serum complement (Quidel, #A113), quickly thawed in a 37°C water bath, was diluted 1:6 in assay medium and added to the assay plate at 50 μL / well. The plate was incubated at 37°C for 2 hours, followed by the addition of 100 μL / well of CellTiter Glo substrate (Promega, #G7571). The plate contents were mixed using a plate shaker at low speed, incubated at room temperature for 5 minutes, and the luminescence signal was read on a BioTek microplate reader (BioTek Instruments) using an integration of 0.2 cps. Data were analyzed using GraphPad Prism 9, and the relative luminescence units (RLU) for each antibody concentration were plotted in a scatter format of antibody concentration vs. RLU. Results are representative of two independent experiments.

[0138] The results, shown in Figure 11, demonstrate that the exemplary IL-4Rα antibodies 5559 IgG1A 124C / 378C, 5559 IgG4P KRQ 124C / 378C, and 5559 IgG4P 124C / 378C did not induce CDC activity when compared to the positive control.

[0139] Example 5. Biophysical properties of human IL-4Rα antibodies The biophysical properties of the exemplified human IL-4Rα antibodies 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P 124C / 378C, and 5559 IgG4P KRQ 124C / 378C were evaluated.

[0140] Aggregation from cell culture: Exemplary antibodies were transiently expressed in CHO cells. Antibody titers and percentages of high molecular weight (% HMW) species after Protein A affinity chromatography purification are listed in Table 12. The results shown in Table 12 indicate that incorporation of an engineered cysteine ​​into the exemplary antibodies or introduction of the KRQ mutation into the exemplary 5559 IgG4P antibody did not significantly affect antibody titer or antibody aggregation.

[0141] Viscosity: Exemplary antibody samples were concentrated to approximately 125 mg / mL in a common formulation buffer matrix at pH 6 containing 5 mM histidine along with excipients. The viscosity of each antibody was measured using a VROC® initium (RheoSense) at 15°C using an average of nine replicate measurements. As shown in Table 12, the results indicate that the 5559 IgG4P KRQ 124C / 378C and 5559 IgG4P GNKRQ 124C / 378C antibodies exhibited significantly improved viscosities of 11.6 cP and 9.6 cP, respectively, compared to 5559 IgG4P 124C / 378C lacking the KRQ amino acid residue substitution. The results further showed that 5559 IgG4P KRQ 124C / 378C and 5559 IgG4P GNKRQ 124C / 378C had viscosities comparable to 5559 IgG1A 124C / 378C. The low viscosity of the exemplified antibodies indicated the desirable development properties of the antibodies.

[0142] Thermal stability: Differential scanning calorimetry (DSC) was used to assess the stability of the exemplified antibodies to thermal denaturation. The thermal melting temperatures of the antibodies in PBS, pH 7.2 buffer are listed in Table 12. Although the thermal transition temperatures for each domain were not fully characterized in either the IgG1 or IgG4P constructs, the data presented in Table 12 and Figures 12A and 12B indicate that the incorporation of engineered cysteines into the exemplified antibodies or the introduction of the KRQ mutation into the exemplified 5559 IgG4P antibody did not negatively impact the thermal stability of the antibodies or alter their structural integrity.

[0143] Aggregation due to temperature stress: The solution stability over time of the exemplified antibodies was evaluated at approximately 100 mg / mL in a common 5 mM histidine pH 6.0 buffer containing excipients. Concentrated samples were incubated at 5°C and 35°C, respectively, for a period of 4 weeks. After incubation, the samples were analyzed for the percentage of high molecular weight (% HMW) species using size exclusion chromatography (SEC). The exemplary results, shown in Table 12, demonstrate that incorporation of an engineered cysteine ​​into the exemplified antibodies or introduction of a KRQ mutation into the exemplified 5559 IgG4P antibody did not affect the aggregation profile of the antibodies over a 4-week period at either 5°C or 35°C. Specifically, the exemplary results demonstrate that the antibodies have comparable solution stability.

[0144] [Table 14] The present invention includes the following aspects. <1> An antibody or antigen-binding fragment thereof that specifically binds to an epitope of human IL-4Rα , wherein the epitope is i.D12, M14, S15, I16, Y37, L39, F41, L42, L43, E 45, H47, T48, C49, I50, E52, H62, L64, M65, D66, D 67, V68, V69, D72, R99, P121, P123, P124, D125, P 192 (amino acid residue positions correspond to SEQ ID NO: 15); ii.D12, M14, S15, I16, Y37, L39, F41, L43, E45, H47, T48, C49, I50, H62, L64, M65, D66, D67, V69, D72, R99, P121, P123, P124, D125 (amino acid residue positions are SEQ ID NO: Corresponding to No. 15); or iii.D12, M14, S15, I16, L39, F41, L42, T48, C49 , I50, E52, H62, L64, M65, D66, D67, V68, V69, D72 , R99, P121, P123, P124, D125, P192 (amino acid residue positions are (corresponding to column number 15) an epitope of human IL-4Rα comprising one or more amino acid residues selected from An antibody or antigen-binding fragment thereof that specifically binds. <2> The epitopes are D12, M14, S15, I16, Y37, L39, T48, C4 9, I50, E52, H62, M65, R99, P121, P123, P124, D12 5, P192, containing one or more amino acid residues selected from <1> The antibody or its An antigen-binding fragment of. <3> The epitope is selected from the group consisting of R99, P121, P123, P124, D125, and P192. comprising one or more selected amino acid residues, <1> ~ <2> or the antibody according to any one of An antigen-binding fragment thereof. <4> The epitope is at least one selected from D66, D67, and D125. containing the amino acid residues <1> The antibody or antigen-binding fragment thereof described in <5> The epitope comprises at least one amino acid residue selected from D66 and D67. include, <1> The antibody or antigen-binding fragment thereof described in <6> The epitope comprises at least one amino acid residue selected from D66 and DI25 Including, <1> The antibody or antigen-binding fragment thereof described in <7> the epitope comprises amino acid residue D66; <1> the antibody or antigen-binding fragment thereof according to Piece. <8> Inhibits the binding of human IL-4 to human IL-4Rα <1> ~ <7> Either The antibody or antigen-binding fragment thereof described above. <9> Inhibits the binding of human IL-13 to human IL-4Rα, <1> ~ <8> Either The antibody or antigen-binding fragment thereof described above. <10> An antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, , a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH is a heavy chain complementary the VL comprises light chain complementarity determining regions HCDR1, HCDR2, and HCDR3; including regions LCDR1, LCDR2, and LCDR3; HCDR1 comprises SEQ ID NO: 1, HCDR2 comprises SEQ ID NO:2, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO:4, LCDR2 comprises SEQ ID NO:5; An antibody or its derivatives that specifically binds to human IL-4Rα, wherein LCDR3 comprises SEQ ID NO: 6. Antigen-binding fragment. <11> the VH comprises SEQ ID NO: 7 and the VL comprises SEQ ID NO: 8; <10> The antibody or is its antigen-binding fragment. <12> An antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, , a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH is a heavy chain complementary the VL comprises light chain complementarity determining regions HCDR1, HCDR2, and HCDR3; including regions LCDR1, LCDR2, and LCDR3; HCDR1 comprises SEQ ID NO: 42, HCDR2 comprises SEQ ID NO:2, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 22; LCDR2 comprises SEQ ID NO:5; An antibody or its derivatives that specifically binds to human IL-4Rα, wherein LCDR3 comprises SEQ ID NO: 6. Antigen-binding fragment. <13> the VH comprises SEQ ID NO: 44 and the VL comprises SEQ ID NO: 45; <12> The anti- or an antigen-binding fragment thereof. <14> The antibody i. a heavy chain (HC) comprising SEQ ID NO: 33 and a light chain (LC) comprising SEQ ID NO: 10; ii. a heavy chain (HC) comprising SEQ ID NO: 35 and a light chain (LC) comprising SEQ ID NO: 10; iii. a heavy chain (HC) comprising SEQ ID NO: 9 and a light chain (LC) comprising SEQ ID NO: 10; iv. a heavy chain (HC) comprising SEQ ID NO: 13 and a light chain (LC) comprising SEQ ID NO: 10; v. a heavy chain (HC) comprising SEQ ID NO: 31 and a light chain (LC) comprising SEQ ID NO: 10; vi. a heavy chain (HC) comprising SEQ ID NO: 37 and a light chain (LC) comprising SEQ ID NO: 10; vii. a heavy chain (HC) comprising SEQ ID NO: 52 and a light chain (LC) comprising SEQ ID NO: 10; viii. A heavy chain (HC) comprising SEQ ID NO: 50 and a light chain (LC) comprising SEQ ID NO: 10, or teeth ix. A heavy chain (HC) comprising SEQ ID NO: 46 and a light chain (LC) comprising SEQ ID NO: 47 Including, <10> ~ <13> 1. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 9. <15> including the human IgG1 isotype, <1> ~ <13> or an antibody thereof Antigen-binding fragment. <16> the antibody comprises an alanine at amino acid residue 322 (EU numbering), <15> Described in An antibody or antigen-binding fragment thereof. <17> including the human IgG4 isotype, <1> ~ <13> or an antibody thereof Antigen-binding fragment. <18> The antibody is selected from the group consisting of: at amino acid residue 137 (EU numbering) glycine , asparagine at amino acid residue 203 (EU numbering); lysine at amino acid residue 274 (EU numbering); an arginine at amino acid residue 355 (EU numbering), or Glutamine at amino acid residue 419 (EU numbering) including one or more of: <17> The antibody or antigen-binding fragment thereof described in <19> The antibody Lysine at amino acid residue 274 (EU numbering), arginine at amino acid residue 355, and and glutamine at amino acid residue 419 (EU numbering). Including, <17> 2. The antibody or antigen-binding fragment according to claim 1. <20> The antibody Glycine at amino acid residue 137 (EU numbering), and 203 (EU numbering) ) asparagine, amino acid residue 274 (EU numbering) lysine, amino acid residue 355 arginine at 419 and glutamine at amino acid residue 419 (EU numbering) Including, <17> 2. The antibody or antigen-binding fragment according to claim 1. <21> An antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, , a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH is a heavy chain complementary the VL comprises light chain complementarity determining regions HCDR1, HCDR2, and HCDR3; including regions LCDR1, LCDR2, and LCDR3; HCDR1 comprises SEQ ID NO: 19, HCDR2 comprises SEQ ID NO: 20, HCDR3 comprises SEQ ID NO: 3, LCDR1 comprises SEQ ID NO: 22; LCDR2 comprises SEQ ID NO:5; An antibody or its derivatives that specifically binds to human IL-4Rα, wherein LCDR3 comprises SEQ ID NO: 24 An antigen-binding fragment of. <22> the VH comprises SEQ ID NO: 25 and the VL comprises SEQ ID NO: 26; <21> The anti- or an antigen-binding fragment thereof. <23> comprising an HC comprising SEQ ID NO: 27 and an LC comprising SEQ ID NO: 28; <21> or <22> of Any of the antibodies or antigen-binding fragments thereof described above. <24> The antibody a cysteine ​​at amino acid residue 124 (EU numbering); a cysteine ​​at amino acid residue 378 (EU numbering), or A cysteine ​​at amino acid residue 124 (EU numbering) and a cysteine ​​at amino acid residue 378 (E cysteine ​​in U numbering), Including, <14> ~ <20> or <23> 1. The antibody or antigen-binding fragment according to any one of claims 1 to 9. <25> SEQ ID NO: 9, 10, 13, 27, 28, 31, 33, 35, 37, 46, 47, 50 or A nucleic acid comprising a sequence encoding 52. <26> <25> A vector comprising the nucleic acid described in . <27> A first nucleic acid encoding SEQ ID NO: 9, 13, 31, 33, 35, 37, 46, 50 or 52 a nucleic acid sequence and a second nucleic acid sequence encoding SEQ ID NO: 10 or 47, <26> Described in Vector of. <28> A first nucleic acid sequence encoding SEQ ID NO:27 and a second nucleic acid sequence encoding SEQ ID NO:28 columns, including <26> The vector described in <29> Nucleic acid sequences encoding SEQ ID NO: 9, 13, 31, 33, 35, 37, 46, 50 or 52 a first vector containing a nucleic acid sequence encoding SEQ ID NO: 10 or 47; and a second vector containing a nucleic acid sequence encoding SEQ ID NO: 10 or 47. A composition comprising: <30> A first vector containing a nucleic acid sequence encoding SEQ ID NO:27 and a second vector containing a nucleic acid sequence encoding SEQ ID NO:28. and a second vector comprising a nucleic acid sequence comprising the nucleic acid sequence. <31> <26> or <28> A cell comprising the vector according to any one of the preceding items. <32> the cell is a mammalian cell; <31> The cell described in <33> <31> ~ <32> and subjecting the cells described in any one of the above to conditions such that the antibody is expressed. and recovering the expressed antibody from the culture medium. A process for producing the antibody or antigen-binding fragment thereof. <34> <33> An antibody or antigen-binding fragment thereof produced by the method described in claim 1. <35> <1> ~ <24> or <34> An antibody-drug conjugate comprising the antibody described in any one of Route. <36> <1> ~ <24> or <34> The antibody or antigen-binding fragment thereof according to any one of the preceding claims, and a drug A pharmaceutical composition comprising a physiologically acceptable excipient, diluent or carrier. <37> Methods for treating an IL-4R-associated disorder in a subject in need thereof a therapeutically effective amount of <1> ~ <24> or <34> The antibody or antibody mixture according to any one of or an antigen-binding fragment thereof, or <36> administering to the subject the pharmaceutical composition described in and treating an IL-4R-associated disorder in a subject in need thereof. method. <38> the IL-4R-associated disorder is an immunoinflammatory disorder; <37> The method described below. <39> the immune inflammatory disorder is a type 2 inflammatory disorder; <38> The method described below. <40> The type 2 inflammatory disorder is atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, Chronic sinusitis (CRS), allergic disease, chronic obstructive pulmonary disease (COPD), or chronic Congenital urticaria (CSU), <39> The method described below. <41> the IL-4R-associated disorder is cancer; <37> The method described below. <42> For use in treatment, <1> ~ <24> or <34> The antibody or its An antigen-binding fragment of. <43> for use in the treatment of IL-4R-associated disorders, <1> ~ <24> or <34> or an antibody or antigen-binding fragment thereof according to any one of <36> The pharmaceutical composition described in <44> the IL-4R-associated disorder is an immunoinflammatory disorder; <43> Antibodies for use as described in or an antigen-binding fragment thereof, or a pharmaceutical composition thereof. <45> the immune inflammatory disorder is a type 2 inflammatory disorder; <44> Antibodies or or an antigen-binding fragment thereof, or a pharmaceutical composition thereof. <46> The type 2 inflammatory disorder is atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, Chronic sinusitis (CRS), allergic disease, chronic obstructive pulmonary disease (COPD) or chronic idiopathic selected from chronic urticaria (CSU), <45> an antibody or its antibody for use according to the A pharmaceutical composition comprising: a hydroxybenzoate; ... <47> the IL-4R-associated disorder is cancer; <43> ANTIBODY OR PHARMACEUTICAL COMPOSITION FOR THE USE OF thing. <48> In the manufacture of a medicament for treating an IL-4R-associated disorder, <1> ~ <24> or < 34> the use of an antibody according to any one of < 34>. <49> the IL-4R-associated disorder is an immunoinflammatory disorder; <48> Use as described in. <50> the immune inflammatory disorder is a type 2 inflammatory disorder; <49> Use as described in. <51> The type 2 inflammatory disorder is atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, Chronic sinusitis (CRS), allergic disease, chronic obstructive pulmonary disease (COPD), or chronic Congenital urticaria (CSU), <50> Use as described in. <52> the IL-4R-associated disorder is cancer; <51> Use as described in.

[0145] Sequence Listing 5559 IgG1A 124C / 378C HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 9 5559 HC for IgG1A 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLV TVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 11 5559 HC DNA for IgG1A 124C / 378C LC DNA for Array No. 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0146] 5559 IgG4P KRQ 124C / 378C HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 13 5559 HC for IgG4P KRQ 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 14 5559 HC DNA for IgG4P KRQ 124C / 378C LC DNA for SEQ ID NO: 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC SEQ ID NO: 15 Human IL-4Rα Extracellular Domain MKVLQEPTCVSDYMSISTCEWKMNGPTNCSTELRLLYQLVFLLSEAHTCIPENNGGAGCVCHLLMDDVVSADNYTLDLWAGQQLLWKGSFKPSEHVKPRAPGNLTVHTNVSDTLLLTWSNPYPPDNYLYNHLTYAVNIWSENDPADFRIYNVTYLEPSLRIAASTLKSGISYRARVRAWAQCYNTTWSEWSPSTKWHNSYREPFEQH SEQ ID NO: 16 Cynomolgus monkey IL-4Rα extracellular domain MKVLQEPTCVSDYMSISTCEWKMGGPTNCSAELRLLYQLVFQSSETHTCVPENNGGVGCVCHLLMDDVVSMDNYTLDLWAGQQLLWKGSFKPSEHVKPRAPGNLTVHTNVSDTVLLTWSNPYPPDNYLYNDLTYAVNIWSENDPAYSRIHNVTYLKPTLRIPASTLKSGISYRARVRAWAQHYNTTWSEWSSPSTKWYNSYREPFEQR SEQ ID NO: 17 human IL-4 MGLTSQLLPPLFFLLACAGNFVHGHKCDITLQEIIKTLNSLTEQKTLCTELTVTDIFAASKNTTEKETFCRAATVLRQFYSHHEKDTRCLGATAQQFHRHKQLIRFLKRLDRNLWGLAGLNSCPVKEANQSTLENFLERLKTIMREKYSKCSS SEQ ID NO: 18 human IL-13 MHPLLNPLLLALGLMALLLTTVIALTCLGGFASPGPVPPSTALRELIEELVNITQNQKAPLCNGSMVWSINLTAGMYCAALESLINVSGCSAIEKTQRMLSGFCPHKVSAGQFSSLHVRDTKIEVAQFVKDLLLHLKKLFREGRFN

[0147] 5F3 IgG4PAA SEQ ID NO: 19 HCDR1 (North) for 5F3 IgG4PAA AASGFTFSISSMN SEQ ID NO: 20 HCDR2 (North) for 5F3 IgG4PAA YISRATGAIY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY SEQ ID NO: 22 LCDR1 (North) for 5F3 IgG4PAA and 8660 IgG4P 124C / 378C RASQGISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 24 LCDR3 (North) for 5F3 IgG4PAA LQHNSYPRT SEQ ID NO: 25 VH for 5F3 IgG4PAA QVQLVESGGGLVQPGGSLRLSCAASGFTFSISSMNWVRQAPGKGLEWVSYISRATGAIYYADSVKGRFTISRNNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 26 VL for 5F3 IgG4PAA DIQMTQSPSAMSASVGDRVTITCRASQGISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPRTFGQGTKVEIK SEQ ID NO: 27 5F3 HC for IgG4PAA QVQLVESGGGLVQPGGSLRLSCAASGFTFSISSMNWVRQAPGKGLEWVSYISRATGAIYYADSVKGRFTISRNNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 28 LC for 5F3 IgG4PAA DIQMTQSPSAMSASVGDRVTITCRASQGISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHNSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 29 HC DNA for 5F3 IgG4PAA LC DNA for Array No. 30 5F3 IgG4PAA GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGGCATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGCATAATAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGAACTGTGGCGGCGCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCCGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0148] 5559 IgG4P 124C / 378C HCDR1 (North) for Array No. 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 31 5559 HC for IgG4P 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 32 5559 HC DNA for IgG4P 124C / 378C LC DNA for Array No. 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0149] 5559 IgG1A HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 33 5559 HC for IgG1A QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 34 5559 HC DNA for IgG1A LC DNA for Array No. 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0150] 5559 IgG4P KRQ HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 35 5559 HC for IgG4P KRQ QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 36 5559 HC DNA for IgG4P KRQ LC DNA for Array No. 125559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0151] 5559 IgG4P HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 37 5559 HC for IgG4P QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 38 5559 HC DNA for IgG4P LC DNA for Array No. 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC SEQ ID NO: 39 Human IL-4Rα MGWLCSGLLFPVSCLVLLQVASSGNMKVLQEPTCVSDYMSISTCEWKMNGPTNCSTELRLLYQLVFLLSEAHTCIPENNGGAGCVCHLLMDDVVSADNYTLDLWAGQQLLWKGSFKPSEHVKPRAPGNLTVHTNVSDTLLLTWSNPYPPDNYLYNHLTYAVNIWSENDPADFRIYNVTYLEPSLRIAASTLKSGISYRARVRAWAQ CYNTTWSEWSPSTKWHNSYREPFEQHLLLGVSCIVILAVCLLCYVSITKIKKEWWDQIPNPARSRLVAIIIQDAQGSQWEKRSRGQEPAKCPHWKNCLTKLLPCFLEHNMKRDEDPHKAAKEMPFQGSGKSAWCPVEISKTVLWPESISVVRCVELFEAPVECEEEEEVEEEKGSFCASPESSRDDFQEGREGIVARLTESLFL DLLGEENGGFCQQDMGESCLLPPSGSTSAHMPWDEFPSAGPKEAPPWGKEQPLHLEPSPPASPTQSPDNLTCTETPLVIAGNPAYRSFSNSLSQSPCPRELGPDPLLARHLEEVEPEMPCVPQLSEPTTVPQPEPETWEQILRRNVLQHGAAAAPVSAPTSGYQEFVHAVEQGGTQASAVVGLGPPGEAGYKAFSSLLASSAVSPE KCGFGASSGEEGYKPFQDLIPGCPGDPAPVPVPLFTFGLDREPPRSPQSSHLPSSSPEHLGLEPGEKVEDMPKPPLPQEQATDPLVDSLGSGIVYSALTCHLCGHLKQCHGQEDGGQTPVMASPCCGCCCGDRSSPPTTPLRAPDPSPGGVPLEASLCPASLAPSGISEKSKSSSSFHPAPGNAQSSSQTPKIVNFVSVGPTYMRVS SEQ ID NO: 40 Cynomolgus monkey IL-4Rα MGWLCSGLLFPVSCLVLLQVASSGCSCVSPGSMKVLQEPTCVSDYMSISTCEWKMGGPTNCSAELRLLYQLVFQSSETHTCVPENNGGVGCVCHLLMDDVVSMDNYTLDLWAGQQLLWKGSFKPSEHVKPRAPGNLTVHTNVSDTVLLTWSNPYPPDNYLYNDLTYAVNIWSENDPAYSRIHNVTYLKPTLRIPASTLKSGISYRARV RAWAQHYNTTWSEWSSPSTKWYNSYREPFEQRLLWGVSAACVFILFFCLSCYFSVTKIKKEWWDQIPNPARSHLVAIIIQDAQESQWEKRSRGQEAAKCPYWKNC LTKLLPCFLEHNMKRDEDPHKAVKDLPFRGSGKSAWCPVEISKTVLWPESISVVRCVELFEAPVECKEEEEVEEEKGSFCTSSESNRDDFQEGREGIVARLTES LFLDLLGGENGGFFQQDMGESCLLPPLGSTSAHVPWDEFPSAGSKEVPPWGKEQPLHQEPSPPASPTQSPDNPTCTEMPLVISSNPAYRSFSNSLSQSPCPRELGPDPLLARHLEEVDPEMPCAPQLSEPTTVAPAEPETWEQILRRNVLQHGAAAAPASAPTSGYREFVHAVQQGGIQASAVAGLGPPGEAGYKAFSSLLASSAVSP GECGFGASSGEEGYKPFQDLTPGCPGDPAPVPVPLFTFGLDREPPHSPQSSHLPSNSPEHLALEPGEKVEDMQKPPLPPEQATDPLGDSLGSGIVYSALTCHLCGHLKQCHGQEDGGQAPVVASPCCGCCCGDRSSPPTTPLRAPDPSLGGVPLEASLCPASLAPSGISEKSKSSLSFHPAPGSAQSSSQTPQIVNFVSVGPTCMRVS SEQ ID NO: 41 human CD23 MEEGQYSEIEELPRRRCCRRGTQIVLLGLVTAALWAGLLTLLLLWHWDTTQSLKQLEERAARNVSQVSKNLESHHGDQMAQKSQSTQISQELEELRAEQQRLKSQDLELSWNLNGLQADLSSFKSQELNERNEASDLLERLREEVTKLRMELQVSSGFVC NTCPEKWINFQRKCYYFGKGTKQWVHARYACDDMEGQLVSIHSPEEQDFLTKHASHTGSWIGLRNLDLKGEFIWVDGSHVDYSNWAPGEPTSRSQGEDCVMMRGSGRWNDAFCDRKLGAWVCDRLATCTPPASEGSAESMGPDSRPDPDGRLPTPSAPLHS

[0152] 8660 IgG4P 124C / 378C SEQ ID NO: 42 8660 HCDR1 (North) for IgG4P 124C / 378C AASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY SEQ ID NO: 22 LCDR1 (North) for 5F3 IgG4PAA and 8660 IgG4P 124C / 378C RASQGISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 44 8660 VH for IgG4P 124C / 378C QVQLVESGGGLVQPGGSLRLSCAASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 45 8660 VL for IgG4P 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQGISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 46 8660 HC for IgG4P 124C / 378C QVQLVESGGGLVQPGGSLRLSCAASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPCVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 47 8660 LC for IgG4P 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQGISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 48 8660 HC DNA for IgG4P 124C / 378C LC DNA for Accession No. 498660 IgG4P 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGGCATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0153] 5559 IgG4P GNKRQ 124C / 378C HCDR1 (North) for Accession No. 15559 IgG1A, 15559 IgG1A 124C / 378C, 15559 IgG4P, 15559 IgG4P 124C / 378C, 15559 IgG4P KRQ, 15559 IgG4P KRQ 124C / 378C, 15559 IgG4P GNKRQ 124C / 378C, and 15559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 50 5559 HC for IgG4P GNKRQ 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTL VTVSSASTKGPCVFPLAPCSRSTSGSTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVNHKPSNTKVDKRVESKYGPP CPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSLG SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 51 5559 HC DNA for IgG4P GNKRQ 124C / 378C LC DNA for Array No. 125559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

[0154] 5559 IgG1AAA 124C / 378C HCDR1 (North) for SEQ ID NO: 1 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C VASGFTFSHSSMN SEQ ID NO: 2: HCDR2 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C YISRATGAVY HCDR3 (North) for SEQ ID NO: 3 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA AREPVFDY LCDR1 (North) for SEQ ID NO: 4 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C RASQDISNYLA LCDR2 (North) for SEQ ID NO: 5 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, 5559 IgG1AAA 124C / 378C, and 5F3 IgG4PAA YAASSLQS SEQ ID NO: 6: LCDR3 (North) for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 8660 IgG4P 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C LQWSSYPRT SEQ ID NO: 7: VH for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLVTVSS SEQ ID NO: 8: VL for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK SEQ ID NO: 52 5559 HC for IgG1AAA 124C / 378C QVQLVESGGGLVQPGGSLRLSCVASGFTFSHSSMNWVRQAPGKGLEWVSYISRATGAVYYADSVKGRFTISRDNAKNSLYLQMNSLRDEDTAVYYCAREPVFDYWGQGTLV TVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 10: LC for 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C DIQMTQSPSAMSASVGDRVTITCRASQDISNYLAWFQQKPGKVPTRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQWSSYPRTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 53 5559 HC DNA for IgG1AAA 124C / 378C LC DNA for Array No. 12 5559 IgG1A, 5559 IgG1A 124C / 378C, 5559 IgG4P, 5559 IgG4P 124C / 378C, 5559 IgG4P KRQ, 5559 IgG4P KRQ 124C / 378C, 5559 IgG4P GNKRQ 124C / 378C, and 5559 IgG1AAA 124C / 378C GACATCCAGATGACCCAGTCTCCATCTGCCATGTCTGCATCTGTGGGAGACAGAGTCACCATCACTTGTCGGGCGAGTCAGGACATTAGCAATTATTTAGCCTGGTTTCAGCAGAAACCAGGGAAAGTCCCTACGCGCCTGATCTATGCTGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACAATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCTACAGTGGTCCAGTTACCCTCGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGC

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, the antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the VH comprising heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3; HCDR1 comprises SEQ ID NO: 1; HCDR2 comprises SEQ ID NO:2; HCDR3 comprises SEQ ID NO:3; LCDR1 comprises SEQ ID NO:4; LCDR2 comprises SEQ ID NO:5; LCDR3 comprises SEQ ID NO: 6; An antibody or an antigen-binding fragment thereof that specifically binds to human IL-4Rα.

2. The antibody or antigen-binding fragment thereof of claim 1, wherein the VH comprises SEQ ID NO: 7 and the VL comprises SEQ ID NO:

8.

3. An antibody or antigen-binding fragment thereof that specifically binds to human IL-4Rα, the antibody comprising a heavy chain variable region (VH) and a light chain variable region (VL), the VH comprising heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprising light chain complementarity determining regions LCDR1, LCDR2, and LCDR3; HCDR1 comprises SEQ ID NO: 42; HCDR2 comprises SEQ ID NO:2; HCDR3 comprises SEQ ID NO:3; LCDR1 comprises SEQ ID NO: 22; LCDR2 comprises SEQ ID NO:5; LCDR3 comprises SEQ ID NO: 6; An antibody or an antigen-binding fragment thereof that specifically binds to human IL-4Rα.

4. The antibody or antigen-binding fragment thereof of claim 3, wherein the VH comprises SEQ ID NO: 44 and the VL comprises SEQ ID NO:

45.

5. The antibody i. a heavy chain (HC) comprising SEQ ID NO: 33 and a light chain (LC) comprising SEQ ID NO: 10; ii. a heavy chain (HC) comprising SEQ ID NO: 35 and a light chain (LC) comprising SEQ ID NO: 10; iii. A heavy chain (HC) comprising SEQ ID NO: 9 and a light chain (LC) comprising SEQ ID NO: 10; iv. a heavy chain (HC) comprising SEQ ID NO: 13 and a light chain (LC) comprising SEQ ID NO: 10; v. a heavy chain (HC) comprising SEQ ID NO: 31 and a light chain (LC) comprising SEQ ID NO: 10; vi. a heavy chain (HC) comprising SEQ ID NO: 37 and a light chain (LC) comprising SEQ ID NO: 10; vii. a heavy chain (HC) comprising SEQ ID NO: 52 and a light chain (LC) comprising SEQ ID NO: 10; viii. a heavy chain (HC) comprising SEQ ID NO: 50 and a light chain (LC) comprising SEQ ID NO: 10, or ix. A heavy chain (HC) comprising SEQ ID NO: 46 and a light chain (LC) comprising SEQ ID NO: 47 The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, comprising:

6. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, comprising a human IgG1 isotype.

7. The antibody or antigen-binding fragment thereof of claim 6, wherein the antibody comprises an alanine at amino acid residue 322 (EU numbering).

8. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 4, comprising a human IgG4 isotype.

9. The antibody is selected from the group consisting of: glycine at amino acid residue 137 (EU numbering), asparagine at amino acid residue 203 (EU numbering); lysine at amino acid residue 274 (EU numbering); an arginine at amino acid residue 355 (EU numbering), or Glutamine at amino acid residue 419 (EU numbering) 9. The antibody or antigen-binding fragment thereof of claim 8, comprising one or more of:

10. The antibody a lysine at amino acid residue 274 (EU numbering), an arginine at amino acid residue 355, and a glutamine at amino acid residue 419 (EU numbering); 9. The antibody or antigen-binding fragment of claim 8, comprising:

11. The antibody a glycine at amino acid residue 137 (EU numbering), an asparagine at amino acid residue 203 (EU numbering), a lysine at amino acid residue 274 (EU numbering), an arginine at amino acid residue 355, and a glutamine at amino acid residue 419 (EU numbering).

9. The antibody or antigen-binding fragment of claim 8, comprising:

12. The antibody a cysteine ​​at amino acid residue 124 (EU numbering), a cysteine ​​at amino acid residue 378 (EU numbering), or a cysteine ​​at amino acid residue 124 (EU numbering) and a cysteine ​​at amino acid residue 378 (EU numbering); The antibody or antigen-binding fragment of any one of claims 5 to 11, comprising: Claim 13: (a) SEQ ID NO: 33 and SEQ ID NO: 10; (b) SEQ ID NO: 9 and SEQ ID NO: 10; (c) SEQ ID NO: 37 and SEQ ID NO: 10; (d) SEQ ID NO: 31 and SEQ ID NO: 10; (e) SEQ ID NO: 35 and SEQ ID NO: 10; (f) SEQ ID NO: 13 and SEQ ID NO: 10; (g) SEQ ID NO: 50 and SEQ ID NO: 10; (h) SEQ ID NO: 52 and SEQ ID NO: 10; or (i) SEQ ID NO: 46 and SEQ ID NO: 47; A vector comprising a nucleic acid sequence encoding 14. (a) a first nucleic acid sequence encoding SEQ ID NO:33, and a second nucleic acid sequence encoding SEQ ID NO:10; (b) a first nucleic acid sequence encoding SEQ ID NO:9 and a second nucleic acid sequence encoding SEQ ID NO:10; (c) a first nucleic acid sequence encoding SEQ ID NO:37 and a second nucleic acid sequence encoding SEQ ID NO:10; (d) a first nucleic acid sequence encoding SEQ ID NO:31 and a second nucleic acid sequence encoding SEQ ID NO:10; (e) a first nucleic acid sequence encoding SEQ ID NO:35 and a second nucleic acid sequence encoding SEQ ID NO:10; (f) a first nucleic acid sequence encoding SEQ ID NO: 13 and a second nucleic acid sequence encoding SEQ ID NO: 10; (g) a first nucleic acid sequence encoding SEQ ID NO:50 and a second nucleic acid sequence encoding SEQ ID NO:10; (h) a first nucleic acid sequence encoding SEQ ID NO:52 and a second nucleic acid sequence encoding SEQ ID NO:10; or (i) a first nucleic acid sequence encoding SEQ ID NO:46, and a second nucleic acid sequence encoding SEQ ID NO:47; The vector of claim 13, comprising:

15. (a) a first vector comprising a nucleic acid sequence encoding SEQ ID NO:33, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10; (b) a first vector comprising a nucleic acid sequence encoding SEQ ID NO:9, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10; (c) a first vector comprising a nucleic acid sequence encoding SEQ ID NO:37, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10; (d) a first vector comprising a nucleic acid sequence encoding SEQ ID NO:31, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10; (e) a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 35, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10; (f) a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 13, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10; (g) a first vector comprising a nucleic acid sequence encoding SEQ ID NO:50, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO:10; (h) a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 52 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10; or (i) a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 46, and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 47; A composition comprising:

16. A cell comprising the vector of claim 13 or 14.

17. The cell of claim 16 , wherein the cell is a mammalian cell.

18. 18. A process for producing an antibody or antigen-binding fragment thereof, comprising culturing a cell described in claim 16 or 17 under conditions such that the antibody is expressed, and recovering the expressed antibody from the culture medium.

19. 20. An antibody or antigen-binding fragment thereof produced by the process of claim 18.

20. 20. An antibody drug conjugate comprising the antibody of any one of claims 1 to 12 or 19.

21. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 12 or 19, and a pharmaceutically acceptable excipient, diluent or carrier.

22. 1. A pharmaceutical product for use in a method of treating an IL-4R-associated disorder in a subject in need thereof, comprising: The pharmaceutical product comprises the antibody or antigen-binding fragment thereof according to any one of claims 1 to 12 or 19, or the pharmaceutical composition according to claim 21; The method comprises administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof described in any one of claims 1 to 12 or 19, or a pharmaceutical composition described in claim 21.

23. The pharmaceutical product of claim 22, wherein the IL-4R-associated disorder is an immunoinflammatory disorder.

24. 24. The pharmaceutical product of claim 23, wherein the immunoinflammatory disorder is a type 2 inflammatory disorder.

25. 25. The pharmaceutical product of claim 24, wherein the type 2 inflammatory disorder is selected from atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic disease, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU).

26. 20. The antibody or antigen-binding fragment thereof of any one of claims 1 to 12 or 19 for use in therapy.

27. The antibody or antigen-binding fragment thereof of any one of claims 1 to 12 or 19, or the pharmaceutical composition of claim 21, for use in treating an IL-4R-associated disorder.

28. The antibody or antigen-binding fragment thereof or pharmaceutical composition for use according to claim 27, wherein the IL-4R-associated disorder is an immunoinflammatory disorder.

29. 29. The antibody or antigen-binding fragment thereof or pharmaceutical composition for use according to claim 28, wherein the immunoinflammatory disorder is a type 2 inflammatory disorder.

30. 30. The antibody or antigen-binding fragment thereof or pharmaceutical composition for use according to claim 29, wherein the type 2 inflammatory disorder is selected from atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU).

31. 20. Use of the antibody or antigen-binding fragment thereof of any one of claims 1 to 12 or 19 in the manufacture of a medicament for treating an IL-4R-associated disorder.

32. The use according to claim 31, wherein the IL-4R-associated disorder is an immunoinflammatory disorder.

33. 33. The use of claim 32, wherein the immunoinflammatory disorder is a type 2 inflammatory disorder.

34. 34. The use of claim 33, wherein the type 2 inflammatory disorder is selected from atopic dermatitis, eosinophilic esophagitis, nasal polyposis, asthma, chronic rhinosinusitis (CRS), allergic diseases, chronic obstructive pulmonary disease (COPD), or chronic spontaneous urticaria (CSU).

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