Modifying binding molecules to minimize pre-existing interactions

Abstract

This disclosure relates to modifying binding molecules to minimize existing binding interactions, including engineering binding molecules to minimize or mitigate background reactivity in a sample matrix caused by nonspecific drug binding interactions.

Description

Engineering Binding Molecules to Minimize Pre-Existing Interactions The present disclosure is directed to engineering binding molecules to minimize pre-existing binding interactions, including binding molecules engineered to minimize or reduce background reactivity in a sample matrix caused by drug non-specific binding interactions. [Related Applications] This application claims the priority and benefit of U.S. Provisional Application No. 62 / 695,988, filed Jul. 10, 2018, the entire content of which is incorporated herein by reference. [Sequence Listing] This application includes a sequence listing that has been submitted in ASCII format via EFS-Web and the entire content of which is incorporated herein by reference. The ASCII copy, created on Jul. 8, 2019, is named “REGE-004_SeqList.txt” and is 32,813 bytes in size. Biotherapeutics are valuable tools for eliminating, supplementing, or replacing an individual's immune system to treat disease. Foreign biological substances have the potential to induce an immune response in the treated individual. The immune response can be triggered by administration of a biotherapeutic agent. However, an individual may have serum protein elements that can generate signals prior to the introduction of the biologic agent, thus creating a high background or noise when tested in an anti-drug antibody (ADA) assay. The standard assay employed during the development and monitoring of biotherapeutic agents is the anti-drug antibody (ADA) assay. This assay is used to detect whether an individual's immune system has produced antibodies against the administered biologic agent. For an effective assay, the signal-to-noise ratio must be a proportion at which meaningful data can be obtained and analyzed. In some individuals, there is a pre-existing high background (noise) even without administration of any biologic agent, which confounds the detection of true treatment-emergent ADA due to the background signal, reducing the effectiveness of the ADA assay. The compositions and methods of the present invention provide a means to reduce or eliminate the drug non-specific, pre-existing background reactivity present in some human serum or plasma samples observed in some ADA assays. It should also be understood that the drug non-specific binding may or may not interfere with the ability of the therapeutic agent to be active or remain active in circulation. The present disclosure is directed to binding molecules engineered to reduce pre-existing drug non-specific background reactivity in an individual sample before or after administration of the binding molecule. The present invention is directed to engineering binding molecules to minimize pre-existing binding interactions, including binding molecules engineered to minimize or reduce background reactivity in a sample matrix caused by drug non-specific binding interactions. The present disclosure further provides binding molecules that are specific to one or more specific targets. In one aspect, the present disclosure is directed to binding molecules that are specific to IL4Rα (interleukin 4α) or IL13R (interleukin 13). In another aspect, the present disclosure provides binding molecules directed to IgG4 antibodies or fragments thereof. As understood herein, a binding molecule is a molecule that specifically interacts with a specific target. Examples of such binding molecules include, but are not limited to, antibodies (including monoclonal antibodies) and fragments thereof, engineered antibodies, fusion proteins, and other similar antigen-binding molecules well known to those skilled in the art. In one aspect, the target is IL4Rα. In another aspect of the present invention, there is disclosed a non-naturally occurring binding molecule comprising a C-terminal heavy chain sequence LSPG (SEQ ID NO: 21) or an antigen-binding portion thereof, which binding molecule is engineered to minimize or reduce background reactivity caused by drug non-specific binding interactions in the sample matrix. The present disclosure provides a non-naturally occurring binding molecule comprising a C-terminal heavy chain sequence SEQ ID NO: 21 or an antigen-binding portion thereof, wherein the binding molecule can reduce interactions with pre-existing serum proteins, and thus can reduce high background signals during ADA assays. In certain embodiments of the present invention, the binding molecule comprises a CH domain sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, or an antigen-binding portion thereof. In certain embodiments, the binding molecule comprises a CH domain sequence selected from the group consisting of SEQ ID NO: 7 and SEQ ID NO: 8, or an antigen-binding portion thereof. In certain embodiments, the binding molecule comprises a CH domain comprising the amino acid sequence of SEQ ID NO: 7. In certain embodiments of the present invention, the binding molecule comprises a truncated CH domain (REGN-E), wherein the sequence is SEQ ID NO: 22. In certain embodiments of the non-naturally occurring binding molecules of the present disclosure, the binding molecule comprises V H CDR1 region, V H CDR2 region, V H CDR3 region, V L CDR1 region, V L CDR2 region, V L CDR3 region and C H3 domains, the V H The CDR1 region contains the amino acid sequence of SEQ ID NO: 9; the V H The CDR2 region contains the amino acid sequence of SEQ ID NO: 10; the V H The CDR3 region contains the amino acid sequence of SEQ ID NO: 11; the V L The CDR1 region contains the amino acid sequence of SEQ ID NO: 12; the V L The CDR2 region contains the amino acid sequence of LGS; the V L The CDR3 region contains the amino acid sequence of SEQ ID NO: 14; the C H The 3 domains are selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7. In certain embodiments of the present invention, the binding molecule comprises V H CDR1 region, V H CDR2 region, V H CDR3 region, V L CDR1 region, V L CDR2 region, V L CDR3 region, and C H 3 domains, the V H The CDR1 region contains the amino acid sequence of SEQ ID NO: 9; the V H The CDR2 region contains the amino acid sequence of SEQ ID NO: 10; the V H The CDR3 region contains the amino acid sequence of SEQ ID NO: 11; the V L The CDR1 region contains the amino acid sequence of SEQ ID NO: 12; the V L The CDR2 region contains the amino acid sequence of LGS; the V LThe CDR3 region contains the amino acid sequence of SEQ ID NO: 14; the C H 3 structure contains the amino acid sequence of SEQ ID NO: 21. The present disclosure provides a non-naturally occurring binding molecule, wherein the binding molecule comprises a V H CDR1 region, a V H CDR2 region, a V H CDR3 region, a V L CDR1 region, a V L CDR2 region, a V L CDR3 region and a C H 3 domain, wherein the V H CDR1 region contains the amino acid sequence of SEQ ID NO: 9; the V H CDR2 region contains the amino acid sequence of SEQ ID NO: 10; the V H CDR3 region contains the amino acid sequence of SEQ ID NO: 11; the V L CDR1 region contains the amino acid sequence of SEQ ID NO: 12; the V L CDR2 region contains the amino acid sequence of LGS; the V L CDR3 region contains the amino acid sequence of SEQ ID NO: 14; the C H 3 domain contains the following amino acid sequences: SEQ ID NO: 5, or SEQ ID NO: 6, or SEQ ID NO: 7, or SEQ ID NO: 21, or SEQ ID NO: 22. In certain embodiments of the present invention, the binding molecule comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 15, and the light chain variable region contains the amino acid sequence of SEQ ID NO: 16. In certain embodiments of the present invention, the binding molecule comprises an IgG4 C H 1 region, the IgG4 C HRegion 1 contains the amino acid sequence of SEQ ID NO: 1. In certain embodiments of the present invention, the binding molecule comprises an IgG4 C H region 2, and the IgG4 C H region 2 contains the amino acid sequence of SEQ ID NO: 2. In certain embodiments of the non-naturally occurring binding molecules of the present disclosure, the binding molecule comprises a hinge region. In certain embodiments, the hinge region comprises the sequence APEFLG (SEQ ID NO: 17). In certain embodiments of the non-naturally occurring binding molecules of the present disclosure, the binding molecule comprises an IgG4 constant region, and the IgG4 constant region contains the amino acid sequence of SEQ ID NO: 8. The present disclosure provides an assay that includes (a) a solid support to which a first component is operably linked; (b) at least one capture agent to which a second component is operably linked, wherein the capture agent comprises a first non-naturally occurring binding molecule, such as a monoclonal antibody, and the first non-naturally occurring binding molecule (such as a monoclonal antibody) comprises sequences encoding V H the CDR1 region, sequences encoding V H the CDR2 region, sequences encoding V H the CDR3 region, sequences encoding V L the CDR1 region, sequences encoding V L the CDR2 region, sequences encoding V L the CDR3 region, and sequences encoding the heavy chain constant region, wherein the heavy chain constant region contains the sequence of SEQ ID NO: 21, and (c) at least one detection agent to which a detectable label is operably linked, wherein the detection agent comprises a second non-naturally occurring binding molecule (such as a monoclonal antibody), and the sequence encoding the detection agent contains sequences encoding the V H CDR1 region of the first non-naturally occurring binding molecule of (b), sequences encoding the V H CDR2 region of the first non-naturally occurring binding molecule of (b), sequences encoding the V HThe sequence of the CDR3 region, the V of the first non-naturally occurring binding molecule encoding (b) L The sequence of the CDR1 region, the V of the first non-naturally occurring binding molecule encoding (b) L The sequence of the CDR2 region, the V of the first non-naturally occurring binding molecule encoding (b) L The sequence of the CDR3 region and wherein the first component and the second component selectively bind to each other. In certain embodiments of the disclosed assay, the first non-naturally occurring binding molecule comprises a sequence encoding a heavy chain variable region and a sequence encoding a light chain variable region, and wherein the second non-naturally occurring binding molecule comprises a sequence encoding the heavy chain variable region of the first non-naturally occurring binding molecule and a sequence encoding the light chain variable region of the first non-naturally occurring binding molecule. In certain embodiments of the disclosed assay, the first non-naturally occurring binding molecule comprises a sequence encoding a heavy chain constant region, the heavy chain constant region comprising SEQ ID NO: 7. In certain embodiments of the disclosed assay, the first non-naturally occurring binding molecule comprises: V H CDR1 region, the V H CDR1 region comprises the amino acid sequence of SEQ ID NO: 9; V H CDR2 region, the V H CDR2 region comprises the amino acid sequence of SEQ ID NO: 10; V H CDR3 region, the V H CDR3 region comprises the amino acid sequence of SEQ ID NO: 11; V L CDR1 region, the V L CDR1 region comprises the amino acid sequence of SEQ ID NO: 12; V L CDR2 region, the V L CDR2 region comprises the amino acid sequence of LGS; V L CDR3 region, the V L CDR3 region comprises the amino acid sequence of SEQ ID NO: 14. In certain embodiments of the disclosed assays, the first non-naturally occurring binding molecule comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 15 and the light chain variable region comprising the amino acid sequence of SEQ ID NO: 16. In one embodiment of the invention, the C H 3 domain (such as dupilumab) of the IgG4 antibody is converted to an IgG1 C H 3 domain. In a further embodiment, the C H 3 domain of the IgG4 antibody (such as dupilumab) is truncated. In one aspect, the truncation occurs at serine 444. In certain embodiments of the disclosed assays, the detection agent comprises dupilumab. In certain embodiments, the second non-naturally occurring binding molecule comprises dupilumab. In certain embodiments of the disclosed assays, the first component comprises streptavidin. In certain embodiments, the second component comprises biotin. The present disclosure provides an assay comprising (a) a solid support, wherein the first component is operably linked to the solid support; (b) at least one capture agent, wherein the second component is operably linked to the at least one capture agent and wherein the capture agent comprises a non-naturally occurring binding molecule or a composition of the present disclosure, and (c) at least one detection agent, wherein a detectable label is operably linked to the detection agent and wherein the detection agent comprises dupilumab; wherein the first component and the second component selectively bind to each other. In certain embodiments, the first component comprises streptavidin. In certain embodiments, the second component comprises biotin. In certain embodiments, a binding molecule that does not specifically bind to the sequence of the variable region of dupilumab does not bind to the at least one capture agent. In certain embodiments, a binding molecule that specifically binds to the sequence of the variable region of dupilumab binds to the at least one capture agent and to at least one detection agent. The present disclosure provides a method for determining the degree of immunogenicity of a biotherapy in an individual, the method comprising (a) contacting a biological sample from the individual with an assay of the present invention under conditions suitable to permit binding of at least one antibody in the biological sample to at least one capture agent and at least one detection agent, wherein prior to the contacting step, the individual has been administered the binding molecule therapy, (b) detecting a signal from the at least one detection agent, and (c) when the signal from (b) is above a threshold, determining that the degree of immunogenicity of the individual is high, or (d) when the signal from (b) is below a threshold, determining that the degree of immunogenicity of the individual is low. Certain embodiments of the present invention are directed to methods for determining the degree of immunogenicity of a biotherapy, wherein the biotherapy comprises a binding molecule as described herein. In one aspect, the biologic agent comprises dupilumab. In particular embodiments of the present invention, a method for determining the degree of immunogenicity of a biotherapy is disclosed, wherein the threshold is a predetermined value. In certain aspects, the threshold is a safety threshold. In certain embodiments of the present invention, the amount of the biotherapy is a therapeutically effective dose, wherein a therapeutically effective dose is an amount of a therapeutic agent (such as a binding molecule of the present invention) that, when administered to an individual, has an amount sufficient to achieve a desired purpose. In some embodiments of the present invention, the degree of immunogenicity is a baseline degree. In certain aspects, the degree of immunogenicity is a subsequent degree or a post-treatment degree. In other embodiments of the present invention, the individual is a participant in a clinical trial. In one aspect, the individual is a patient receiving medical treatment. In another aspect, the medical treatment is beginning and the degree of immunogenicity is a baseline degree. In yet another aspect, the medical treatment is ongoing and the degree of immunogenicity is a subsequent degree. In yet another aspect, the medical treatment is ending and the degree of immunogenicity is a final degree. In another aspect, the individual is a healthy individual. In still other embodiments of the present invention, the individual has an inflammatory disease or disorder, an autoimmune disease or disorder, an allergic disease or disorder, an immune disease or disorder, or a benign proliferative disease or disorder. In one aspect, the individual has atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis, nasal polyps, ABPA (allergic bronchopulmonary aspergillosis), bullous pemphigoid, chronic obstructive pulmonary disease (COPD), HFE (hand-foot eczema), prurigo nodularis, or any type 2 inflammatory response or a combination thereof. The individual may have any disease or medical condition. The present disclosure provides non-naturally occurring monoclonal antibodies that comprise a C-terminal heavy chain sequence that comprises a sequence selected from the group consisting of SEQ ID NO: 7 and SEQ ID NO: 13, or an antigen-binding portion thereof. The C-terminal heavy chain sequence can be a C H 3 domain sequence. The C-terminal heavy chain sequence can comprise SEQ ID NO: 7. The C-terminal heavy chain sequence can comprise SEQ ID NO: 13. The C-terminal heavy chain sequence can comprise C H domain sequence, the C H domain sequence comprises SEQ ID NO: 8. The C-terminal heavy chain sequence can comprise C H domain sequence, the C H domain sequence comprises SEQ ID NO: 22. A non-naturally occurring monoclonal antibody that comprises a C H 3 domain, the C H 3 domain is composed of a sequence selected from the group consisting of SEQ ID NO: 7 and SEQ ID NO: 13, or an antigen-binding portion thereof. A non-naturally occurring monoclonal antibody that comprises a C-terminal heavy chain sequence that is composed of a sequence selected from the group consisting of SEQ ID NO: 8 and SEQ ID NO: 22, or an antigen-binding portion thereof. A non-naturally occurring monoclonal antibody, wherein the antibody comprises: V H CDR1 region, the V H CDR1 region comprises the amino acid sequence of SEQ ID NO: 9; V H CDR2 region, the V H CDR2 region comprises the amino acid sequence of SEQ ID NO: 10; V H CDR3 region, the V H CDR3 region comprises the amino acid sequence of SEQ ID NO: 11; V L CDR1 region, the V LThe CDR1 region contains the amino acid sequence of SEQ ID NO: 12; V L CDR2 region, the V L CDR2 region contains the amino acid sequence of LGS; V L CDR3 region, the V L CDR3 region contains the amino acid sequence of SEQ ID NO: 14; and C H 3 domain, the C H 3 domain is composed of an amino acid sequence selected from the group consisting of: SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 13, or an antigen-binding portion thereof. The aforementioned antibody may further comprise: V H CDR1 region, the V H CDR1 region contains the amino acid sequence of SEQ ID NO: 9; V H CDR2 region, the V H CDR2 region contains the amino acid sequence of SEQ ID NO: 10; V H CDR3 region, the V H CDR3 region contains the amino acid sequence of SEQ ID NO: 11; V L CDR1 region, the V L CDR1 region contains the amino acid sequence of SEQ ID NO: 12; V L CDR2 region, the V L CDR2 region contains the amino acid sequence of LGS; V L CDR3 region, the V L CDR3 region contains the amino acid sequence of SEQ ID NO: 14. This disclosure provides non-naturally occurring monoclonal antibodies, wherein the antibody comprises: V H CDR1 region, the V H The CDR1 region contains the amino acid sequence of SEQ ID NO: 9; V H The CDR2 region, the V H The CDR2 region contains the amino acid sequence of SEQ ID NO: 10; V H The CDR3 region, the V H The CDR3 region contains the amino acid sequence of SEQ ID NO: 11; V L The CDR1 region, the V L The CDR1 region contains the amino acid sequence of SEQ ID NO: 12; V L The CDR2 region, the V L The CDR2 region contains the amino acid sequence of LGS; V L The CDR3 region, the V L The CDR3 region contains the amino acid sequence of SEQ ID NO: 14; and C H The C3 domain, the C H The C3 domain contains an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 13, or an antigen-binding portion thereof. The aforementioned antibody may comprise a heavy chain variable region and a light chain variable region, or an antigen-binding portion thereof, the heavy chain variable region containing the amino acid sequence of SEQ ID NO: 15, and the light chain variable region containing the amino acid sequence of SEQ ID NO: 16. The aforementioned antibody may comprise an IgG4 C H The C1 region, the IgG4 C H The C1 region contains the amino acid sequence of SEQ ID NO: 1. The aforementioned antibody may comprise an IgG4 C H The C2 region, the IgG4 C H The C2 region contains the amino acid sequence of SEQ ID NO: 2. IgG4 C H The C2 region may comprise a hinge region. The hinge region may comprise the sequence of APEFLG (SEQ ID NO: 17). The antibodies of the present disclosure can reduce high background signals during immunogenicity analysis. The present disclosure provides an assay comprising: (a) a solid support to which a first component is operably linked; (b) at least one capture agent to which a second component is operably linked, wherein the capture agent comprises a first non-naturally occurring monoclonal antibody as recited in any one of claims 1 to 14, and (c) at least one detection agent to which a detectable label is operably linked, wherein the detection agent comprises a second non-naturally occurring monoclonal antibody as recited in any one of claims 1 to 11, and wherein the first component and the second component selectively bind to each other. The detection agent may comprise dupilumab. The second non-naturally occurring monoclonal antibody may comprise dupilumab. The first component may comprise streptavidin. The second component may comprise biotin. The present disclosure provides an assay comprising: (a) a solid support to which a first component is operably linked; (b) at least one capture agent to which a second component is operably linked and wherein the capture agent comprises a non-naturally occurring monoclonal antibody as recited in any one of claims 1 to 14; and (c) at least one detection agent to which a detectable label is operably linked and wherein the detection agent comprises dupilumab; wherein the first component and the second component selectively bind to each other. The first component may comprise streptavidin. The second component may comprise biotin. In some aspects, the at least one capture agent does not bind to an antibody that does not specifically bind to the variable region sequence of dupilumab. In some aspects, the at least one capture agent and the at least one detection agent bind to an antibody that specifically binds to the variable region sequence of dupilumab. The present disclosure provides a method for determining the degree of immunogenicity of a monoclonal antibody therapy in an individual, comprising: (a) contacting a biological sample from the individual with an assay as recited in any one of claims 15 to 24 under conditions suitable to allow at least one antibody in the biological sample to bind to at least one capture agent and at least one detection agent, wherein the individual has been administered the monoclonal antibody therapy prior to the contacting step; (b) detecting a signal from the at least one detection agent, and (c) when the signal from (b) is above a threshold, determining that the degree of immunogenicity of the individual is high; or (d) when the signal from (b) is below a threshold, determining that the degree of immunogenicity of the individual is low. The monoclonal antibody therapy may comprise the antibodies of the present disclosure. The monoclonal antibody therapy may include dupilumab. The threshold value may be a predetermined value. The threshold value may be a safety threshold value. The amount of the monoclonal antibody therapy may be a therapeutically effective dose. The degree of immunogenicity may be a baseline degree. The degree of immunogenicity may be a subsequent degree. The individual may be a participant in a clinical trial. The individual may be a patient receiving medical treatment. The medical treatment may be starting and the degree of immunogenicity may be a baseline degree. The medical treatment may be ongoing and the degree of immunogenicity may be a subsequent degree. The medical treatment may be ending and the degree of immunogenicity may be a final degree. The individual may be a healthy individual. The individual may have an inflammatory disease or disorder, an autoimmune disease or disorder, an allergic disease or disorder, an immune disease or disorder, or a benign proliferative disease or disorder. The individual may have atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis, nasal polyps, or a combination thereof. Any of the above aspects may be combined with any other aspect. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. In this patent specification, unless the context clearly dictates otherwise, the singular forms also include the plural forms; for example, the terms "a", "an", and "the" should be understood as singular or plural, and the term "or" should be understood as inclusive. For example, "element" refers to one or more elements. Throughout this patent specification, the word "comprising" or variations such as "comprises" or "comprising" will be understood to imply the inclusion of the specified element, integer, or step, or group of elements, integers, or steps, without excluding any other element, integer, or step, or group of elements, integers, or steps. "About" can be understood to be within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless the context clearly dictates otherwise, all numerical values provided herein are modified by the term "about". It should be understood that this disclosure is not limited to the compositions and methods and the experimental conditions described herein, as these may vary. It should also be understood that the terms used herein are only for describing certain embodiments and are not intended to be limiting, as the scope of this disclosure is only limited by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. However, any compositions, methods, and materials similar or equivalent to those described herein may be used to practice or test the invention. The entire contents of all publications mentioned are incorporated herein by reference. As used herein, the term "human IL4R" (hIL-4R) refers to the human cytokine receptor that specifically binds to interleukin-4 (IL-4), IL-4Rα (SEQ ID NO: 18). The term "human interleukin-13" (hIL-13) refers to the cytokine that specifically binds to the IL-13 receptor, and the "hIL-13 / hIL-13R1 complex" refers to the complex formed by the binding of hIL-13 to the hIL-13R1 complex, which binds to the hIL-4 receptor to initiate biological activity. As used herein, the term "binding molecule" refers to a molecule that specifically interacts with and binds to a specific target. The target can include biological or small (chemical) molecules. The target molecule can define an antigen or antigenic portion. Examples of binding molecules include, but are not limited to, antibodies (including monoclonal antibodies, bispecific antibodies, and antibody fragments), fusion proteins, and other antigen-binding molecules known to those skilled in the art. The term "antibody" as used herein is an example of a binding molecule and refers to an immunoglobulin that typically comprises four polypeptide chains (two heavy (H) chains and two light (L) chains interconnected by disulfide bonds). Each heavy chain comprises a heavy chain variable region (HCVR or V H ) and a heavy chain constant region. The heavy chain constant region comprises three domains, C H 1, C H 2, and C H 3. Each light chain comprises a light chain variable region (LCVR or V L ) and a light chain constant region. The light chain constant region comprises one domain (CL1). The V H and V L regions can be further subdivided into hypervariable regions, called complementarity determining regions (CDRs), which are interspersed with more conserved regions, called framework regions (FRs). Each V H and V L is composed of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Antibodies can include antibodies of the IgG1, IgG2, IgG3, or IgG4 subclasses. Antibodies can also comprise combinations of regions from different subclasses. IgG4 antibodies can include, but are not limited to, dupilumab and cemiplimab. Other examples of "binding molecules" include, but are not limited to, bispecific antibodies, trispecific antibodies, tetraspecific antibodies, and pentaspecific antibodies. In some aspects, the bispecific antibodies, trispecific antibodies, tetraspecific antibodies, and pentaspecific antibodies may comprise the Fc portion of an antibody. In some aspects, the bispecific antibodies, trispecific antibodies, tetraspecific antibodies, and pentaspecific antibodies may comprise an IgG4 backbone. Another example of a "binding molecule" is an antibody-drug conjugate (ADC). In some aspects, the ADC may comprise an IgG4 backbone. Another example of a "binding molecule" is a bispecific T cell engager (BiTE). In some aspects, the BiTE may comprise an IgG4 backbone. Another example of a "binding molecule" is a TRAP fusion protein. In some aspects, the TRAP fusion protein has an IgG4 backbone. Another example of a "binding molecule" is a fynomer. As used herein, the "antigen-binding portion" of an antibody (or simply, "antibody portion" or "antibody fragment") refers to one or more antibody fragments that retain the ability to specifically bind to an antigen (such as hIL-4Rα). The antigen-binding function of an antibody has been shown to be performed by fragments of the full-length antibody. Examples of binding fragments included within the term "antigen-binding portion" of an antibody include (i) Fab fragments, comprising the V L 、V H 、C L 1 and C H 1 domains of a monovalent fragment; (ii) F(ab’)2 fragments, bivalent fragments comprising two F(ab)’ fragments linked by a disulfide bridge in the hinge region; (iii) Fc fragments comprising the V H and C H 1 domains; (iv) Fv fragments comprising the V L and V H domains of a single arm of an antibody; (v) dAb fragments (Ward et al. (1989) Nature 241:544-546), which comprise the V H domain; and (vi) CDRs. In addition, although the two domains of the Fv fragment, V L and V Hare encoded by different genes, which can be joined by recombinant methods using a synthetic linker that allows them to be made into a single continuous chain, where the V L and V H regions pair to form a monovalent molecule (referred to as a single-chain Fv (scFv); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883. These single-chain antibodies are also intended to be included in the "antigen-binding portion" of the term antibody. Other forms of single-chain antibodies, such as diabodies (see, e.g., Holliger et al. (1993) Proc. Natl. Acad Sci. USA 90:6444-6448) are also included. As used herein, a "neutralizing" or "blocking" antibody refers to an antibody that binds to hIL-4Rα and can result in inhibition of the biological activity of hIL-4 and / or hIL-13. Inhibition of the biological activity of hIL-4 and / or IL-13 can be evaluated by measuring one or more indicators of hIL-4 and / or hIL-13 biological activity known in the art, such as cell activation induced by hIL-4 and / or IL-13 and hIL-4 binding to hIL-4Rα (see the following examples). "CDR" or complementarity-determining region is a hypervariable region that is interspersed within more conserved regions, called "framework regions" (FR). In different embodiments of the anti-hIL-4Rα antibodies or fragments of the present disclosure, the FR can be identical to the human germline sequence or can be natural or artificially modified. The term "epitope" is an antigenic determinant that interacts with the specific antigen-binding site in the variable region of an antibody molecule, called a paratope. A single antigen can have more than one epitope. Epitopes can be conformational or linear. Conformational epitopes are produced by amino acids that are spatially juxtaposed from different segments of a linear polypeptide chain. Linear epitopes are epitopes produced by adjacent amino acid residues in a polypeptide chain. In some cases, an epitope can include a sugar, phosphoryl, or sulfonyl moiety on the antigen. The term "immunogenicity" refers to the ability of an antigen or immunogen to induce an immune response in a human or animal. Protein therapeutics have the ability to elicit adverse immune responses that can interfere with the pharmacokinetics and efficacy of the drug. The immune response can be in the form of the production of anti-drug antibodies (ADA). When referring to a nucleic acid or a fragment thereof, the term "substantial identity" or "substantially identical" means that when optimally aligned with another nucleic acid (or its complementary strand) with appropriate nucleotide insertions or deletions, as measured by any well-known sequence identity algorithm such as FASTA, BLAST, or Gap discussed below, it has nucleotide sequence identity in at least about 95%, more preferably at least about 96%, 97%, 98%, or 99% of the nucleotide bases. When applied to polypeptides, the term "substantial similarity" or "substantially similar" means that when two peptide sequences are optimally aligned (such as using default gap weights, by the programs GAP or BESTFIT), they share at least 95% sequence identity, even more preferably at least 98% or 99% sequence identity. Generally, the positions of the non-identical residues differ from each other due to conservative amino acid substitutions. "Conservative amino acid substitution" is one in which an amino acid residue is replaced by another amino acid residue having a side chain (R group) with similar chemical properties (such as charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially change the functional properties of the protein. In cases where two or more amino acid sequences differ from each other due to conservative substitutions, the percentage of sequence identity or degree of similarity can be adjusted upward to correct for the conservative nature of the substitution. Methods for making this adjustment are well known to those skilled in the art. See, for example, Pearson (1994) Methods Mol. Biol. 24: 307-331 (which is incorporated herein by reference). Examples of groups of amino acids with side chains having similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartic acid and glutamic acid, and (7) sulfur-containing side chains: cysteine and methionine. In certain embodiments, conservative amino acid substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative substitution is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443-45 (which is incorporated herein by reference). A "moderately conservative" substitution is any change having a non-negative value in the PAM250 log-likelihood matrix. The sequence similarity of polypeptides (which is also referred to as sequence identity) is typically measured using sequence analysis software. Protein analysis software uses similarity measures attributable to various substitutions, deletions, and other modifications, including conservative amino acid substitutions, to match similar sequences. For example, GCG software contains programs such as Gap and Bestfit, which can be used with default parameters to determine the sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different biological species, or the sequence homology or sequence identity between a wild-type protein and its mutant. See, for example, GCG Version 6.1. Polypeptide sequences can also be compared using the program FASTA in GCG Version 6.1, which uses default or recommended parameters. FASTA (e.g., FASTA2 and FASTA3) provides alignments and the percentage of sequence identity between the best overlapping regions of the query and search sequences (Pearson (2000), supra). Another algorithm for comparing the sequences of the present disclosure with databases containing a large number of sequences from different organisms is the computer program BLAST, particularly BLASTP or TBLASTN, which uses default parameters. See, for example, Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402 (each of which is incorporated herein by reference). Methods for generating human antibodies include, for example, those described in U.S. Patent No. 6,596,541, Green et al. (1994) Nature Genetics 7:13-21, U.S. Patent No. 5,545,807, 6,787,637. Rodents can be immunized by any method known in the art (see, e.g., Harlow and Lane (1988) Antibodies: A Laboratory Manual 1988 Cold Spring Harbor Laboratory; Malik and Lillehoj (1994) Antibody Techniques, Academic Press, CA). The antibodies of the present disclosure are typically prepared using the VELOCIMMUNE® technology (U.S. Patent No. 6,596,541). Transgenic mice in which the endogenous immunoglobulin heavy and light chain variable regions are replaced with the corresponding human variable regions are challenged with the desired antigen, and lymphocytes (such as B cells) are recovered from the mice that express the antibody. The lymphocytes can be fused with a myeloma cell line to prepare immortalized hybridoma cell lines, and the hybridoma cell lines are screened and selected to identify hybridoma cell lines that can produce antibodies specific for the desired antigen. The DNA encoding the heavy and light chain variable regions can be isolated and ligated to the required isotype heavy and light chain constant regions. The antibody proteins can be produced in cells (such as CHO cells). Alternatively, the DNA encoding the antigen-specific chimeric antibody or the variable regions of the light and heavy chains can be directly isolated from antigen-specific lymphocytes. The DNA encoding the variable regions of the heavy and light chains of the antibody can be isolated and operably linked to the DNA encoding the human heavy and light chain constant regions. The DNA can then be expressed in cells capable of expressing a fully human antibody. In a particular embodiment, the cells are CHO cells. The antibody can be therapeutically useful for blocking ligand-receptor interactions or inhibiting receptor component interactions, rather than killing cells by fixing complement (complement-dependent cytotoxicity) (CDC) and participating in antibody-dependent cell-mediated cytotoxicity (ADCC). The constant region of the antibody is important for the ability of the antibody to fix complement and mediate cell-dependent cytotoxicity. Therefore, the isotype of the antibody can be selected based on whether the antibody mediates cytotoxicity. Human immunoglobulins can exist in two forms that are related to hinge heterogeneity. In one form, the immunoglobulin molecule comprises a stable four-chain construct of about 150-160 kDa, wherein the dimer is held together by interchain heavy chain disulfide bonds. In the second form, the dimer is not linked by interchain disulfide bonds and forms a molecule (half-antibody) of about 75-80 kDa, composed of covalently coupled light and heavy chains. These forms are extremely difficult to separate even after affinity purification. The frequency of the second form in the various intact IgG isotypes is due to, but not limited to, structural differences related to the isotype of the hinge region of the antibody. In fact, a single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the occurrence of the second form (Angal et al. (1993) Molecular Immunology 30:105) to the extent typically observed with the human IgG1 hinge. The present disclosure encompasses antibodies having one or more mutations in the hinge, C H 2 or C H 3 region, which mutations may be, for example, those required to increase the yield of the desired antibody form. First, a high-affinity chimeric antibody having a human variable region and a murine constant region is isolated. As described below, antibodies having the desired characteristics (including binding affinity for hIL-4Rα, the ability to block the binding of hIL-4 to hIL-4Rα, and / or selectivity for human proteins) are characterized and selected. The murine constant region is replaced with the desired human constant region to generate the fully human antibodies of the present disclosure, such as wild-type or modified IgG4 or IgG1 (e.g., SEQ ID NO: 4, 19, 20, and 23). Although the constant region selected may vary according to the particular use, the high-affinity antigen-binding and target specificity characteristics reside in the variable region. Immunogenicity analysis The present disclosure provides non-naturally occurring binding molecules (such as monoclonal antibodies or antigen-binding portions thereof) comprising a C-terminal heavy chain sequence LSPG (SEQ ID NO: 21). In certain embodiments, the C-terminal heavy chain is a human C H 3 domain. In certain embodiments, the human antibody belongs to the IgG4 class. In certain embodiments, the heavy chain sequence comprises SEQ ID NO: 7. In certain embodiments, the heavy chain sequence comprises SEQ ID NO: 8. The present disclosure provides immunogenicity assays that include anti-IL-4Rα binding molecules, such as the antibodies or antigen-binding fragments thereof of the present disclosure. The immunogenicity assays of the present disclosure can be in the form of anti-drug antibody (ADA) assays. The ADA assays of the present disclosure can be ADA bridging assays or direct enzyme-linked immunosorbent (ELISA) assays. In the ADA bridging assay, a biotinylated form of the binding molecule in question is bound to streptavidin on a plate. Then, the binding molecule-like antibodies present in the sample bind simultaneously to the biotinylated binding molecule and a labeled form of the same binding molecule, forming a bridging interaction with the detectable signal from the label. Suitable labels are known to those of ordinary skill in the art. Exemplary labels include ruthenium, horseradish peroxidase, alkaline phosphatase, and fluorophores. The ADA bridging assay can include titrating the sample undergoing the bridging reaction to generate a standard curve and using cold, unlabeled competing antibodies to inhibit the reaction. The drug-specific bridging ADA assay is illustrated in Panel A of FIG. 4; the non-drug-specific bridging assay due to pre-existing reactivity in current ADA assays is illustrated in Panel B; and the non-drug-specific bridging ADA assay due to the use of REGN-C as a capture agent is illustrated in Panel C. In Panels B and C, the pre-existing reactivity is represented by the smaller dashed-outline cross structures. It should be understood that in embodiments of the disclosure, the assay reagent molecule analogs REGN-A, B, C, D, and E are interchangeable, thus allowing those of skill in the art to design various assay arrangements as described. In one aspect, to reduce background, those of skill in the art can combine REGN-D with any of the following reagents to form novel combinations: REGN-B, REGN-C, or REGN-E (D + B, D + C, D + E, or reversed to B + D, C + D, or E + D), such that the alignment order is "capture agent" + "detection agent". In another aspect, those of skill in the art can make the following pairings to reduce background signal: B + B, C + C, or E + E. In certain embodiments of the present disclosure, the degree of immunogenicity (or ADA) is determined using a binding molecule, such as an antibody (or antigen-binding portion thereof). In certain embodiments, the degree of immunogenicity of dupilumab is evaluated using IgG4 antibodies. One problem that is typically addressed in these immunogenicity studies is high background signal, which is typically associated with the IgG4 antibodies used. To reduce this high background, the present disclosure provides IgG4 antibodies that include any of the following heavy chain sequences to result in lower background signal: SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 7, or SEQ ID NO: 8. In certain embodiments of the present disclosure, the binding molecule (such as an antibody or an antigen-binding portion thereof) comprises (1) one or more V L chain sequences selected from the group consisting of: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V H chain sequences selected from the group consisting of: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) a C H 3 or CH sequence selected from the group consisting of: SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 7, and SEQ ID NO: 8, wherein the binding molecule shows reduced background reactivity in an immunogenicity (ADA) assay compared to a binding molecule comprising (1) one or more V L chain sequences selected from the group consisting of: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V H chain sequences selected from the group consisting of: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) a C H 3 or CH sequence not selected from the group consisting of: SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 7, and SEQ ID NO: 8. In certain embodiments, the CH domain sequence of (3) may include SEQ ID NO: 5 and SEQ ID NO: 6. In one embodiment of the present invention, the C H 3 domain of the IgG4 antibody (such as dupilumab) is converted to the IgG1 C H 3 domain resulting in a lower background signal (Figure 6B). In a further embodiment, the C H 3 domain of the IgG4 antibody (such as dupilumab) is truncated. In one aspect, the truncation occurs at serine 444 resulting in a lower background signal (Figure 6C). In some aspects, the present disclosure provides a binding molecule comprising (1) one or more V L Heavy chain sequences: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V selected from the group consisting of H Heavy chain sequences: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) C selected from the group consisting of H 3 or C H Sequences: SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 13, SEQ ID NO: 7, and SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising: (1) one or more V selected from the group consisting of L Heavy chain sequences: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V selected from the group consisting of H Heavy chain sequences: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) C H 3 or C H sequence, wherein the C H 3 or C H sequence comprises one or more sequences selected from the group consisting of SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 13, SEQ ID NO: 7, and SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising (1) one or more V selected from the group consisting of L Heavy chain sequences: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V selected from the group consisting of H Heavy chain sequences: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) C H 3 sequence, wherein the C HThe 3 sequence comprises one or more sequences selected from the group consisting of: SEQ ID NO: 7, , SEQ ID NO: 13 or SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising (1) one or more V L chain sequences selected from the group consisting of: SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V H chain sequences selected from the group consisting of: SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) a C H sequence, wherein the C H sequence comprises one or more sequences selected from the group consisting of: SEQ ID NO: 8, SEQ ID NO: 21, and SEQ ID NO: 22. In some aspects, the present disclosure provides a binding molecule comprising a C H sequence, wherein the C H sequence comprises one or more sequences selected from the group consisting of: SEQ ID NO: 8, SEQ ID NO: 21, and SEQ ID NO: 22. In some aspects, the present disclosure provides a binding molecule comprising a C H 3 sequence, wherein the C H 3 sequence comprises one or more sequences selected from the group consisting of: SEQ ID NO: 7, SEQ ID NO: 13, and SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising an IgG4 C H domain. The IgG4 C H domain comprises an amino acid substitution wherein the proline at position 445 is replaced with leucine. In some aspects, the present disclosure provides a binding molecule comprising SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 5. In some aspects, the present disclosure provides a binding molecule comprising SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 6. In some aspects, the present disclosure provides a binding molecule comprising (1) one or more V L chain sequences selected from the group consisting of SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V H chain sequences selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) a C H sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 5. In some aspects, the present disclosure provides a binding molecule comprising (1) one or more V L chain sequences selected from the group consisting of SEQ ID NO: 12, LGS, SEQ ID NO: 14, and combinations thereof, (2) one or more V H chain sequences selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and combinations thereof, and (3) a C H sequence comprising SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 6. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 99% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 98% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 97% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 96% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 95% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 94% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 93% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 92% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 91% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 90% sequence identity to SEQ ID NO: 7. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 99% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 98% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 97% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 96% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 95% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 94% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 93% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 92% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 91% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 90% sequence identity to SEQ ID NO: 8. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 99% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 98% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 913% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 96% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 95% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 94% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 93% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 92% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 91% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 90% sequence identity with SEQ ID NO: 13. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 99% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 98% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 97% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 96% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 95% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 94% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 93% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 92% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 91% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 90% sequence identity with SEQ ID NO: 7, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 99% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 98% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 97% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 96% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 95% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 94% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 93% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 92% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 91% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, the present disclosure provides a binding molecule comprising at least one polypeptide sequence having at least about 90% sequence identity to SEQ ID NO: 8, wherein the at least one polypeptide sequence comprises SEQ ID NO: 21. In some aspects, compared to a binding molecule comprising a C H sequence containing SEQ ID NO: 3, any binding molecule of the present disclosure may show reduced background reactivity in an immunogenicity (ADA) assay. In some aspects, compared to a binding molecule comprising a C HCompared to the binding molecules of the sequences, any binding molecule of the present disclosure may show reduced background reactivity in an immunogenicity (ADA) assay. Therapeutic Administration and Formulations The present disclosure provides therapeutic compositions comprising the anti-IL-4Rα binding molecules of the present disclosure. When administering the therapeutic compositions according to the present disclosure, they will be administered together with suitable carriers, excipients, and other agents incorporated into the formulations to provide improved transfer, delivery, tolerability, etc. Many suitable formulations can be found in the formulations known to all pharmaceutical chemists: Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, vesicles containing lipids (cationic or anionic) such as LIPOFECTIN™, DNA conjugates, anhydrous absorbent pastes, water-in-oil and oil-in-water emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al. “Compendium of excipients for parenteral formulations” PDA (1998) J Pharm Sci Technol 52:238-311. The dosage may vary depending on the age and size of the individual to whom it is to be administered, the target disease, condition, route of administration, etc. When using the binding molecules of the present disclosure to treat various conditions and diseases associated with IL-4Rα in adult patients, it is advantageous to administer the binding molecules of the present disclosure via the intravenous route, usually as a single dose of about 0.01 to about 20 mg / kg body weight, more often as a single dose of about 0.02 to about 7, about 0.03 to about 5, or about 0.05 to about 3 mg / kg body weight. In some aspects, when using the binding molecules of the present disclosure to treat various conditions and diseases associated with IL-4Rα, the dosing regimen may be to administer 300 mg once every two weeks (Q2W) and can be extended to once every four weeks (Q4W). The frequency and duration of treatment can be adjusted according to the severity of the condition. A variety of delivery systems are known and can be used to administer the pharmaceutical compositions of the present disclosure, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant virus, receptor-mediated endocytosis (see, e.g., Wu et al. (1987) J. Biol. Chem. 262:4429-4432). Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The compositions can be administered by any convenient route, for example, by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (such as oral mucosa, rectal, and intestinal mucosa, etc.) and can be administered together with other bioactive agents. Administration can be systemic or local. The pharmaceutical compositions can also be delivered in vesicles, such as liposomes (see Langer (1990) Science 249:1527-1533; Treat et al. (1989) in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez Berestein and Fidler (eds.), Liss, New York, pp. 353-365; Lopez-Berestein, ibid., pp. 317-327). In certain embodiments, the pharmaceutical compositions can be delivered in a controlled release system. In one embodiment, a pump can be used (see Langer, supra; Sefton (1987) CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974)). In yet another embodiment, the controlled release system can be placed near the target of the composition, so that only a fraction of the systemic dose is required (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138, 1984). Discussion of other controlled release systems is provided in the review by Langer (1990) Science 249:1527-1533). The injection preparation may include dosage forms for intravenous, subcutaneous, intradermal, and intramuscular injections, drip infusion, etc. These injection preparations can be prepared by known methods. For example, the injection preparation can be prepared, for example, by dissolving or suspending the above-mentioned antibody or its salt in a sterile aqueous medium or an oily medium commonly used for injection or emulsifying it in a sterile aqueous medium or an oily medium commonly used for injection. Examples of aqueous media that can be used for injection include, for example, physiological saline, isotonic solutions containing glucose and other adjuvants, etc., which can be used in combination with appropriate solubilizers, such as alcohols (e.g., ethanol), polyhydric alcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 moles) adduct of hydrogenated castor oil)], etc. Examples of oily media that can be used include, for example, sesame oil, soybean oil, etc., which can be used in combination with solubilizers, such as benzyl benzoate, benzyl alcohol, etc. The injection preparation thus prepared is preferably filled in appropriate ampoules. Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared in unit dosage forms that are suitable for dosing the active ingredient. These unit dosage forms include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The content of the foregoing binding molecule is usually about 5 to 500 mg per unit dosage form; especially in the injectable form, in certain embodiments, the content of the foregoing binding molecule is about 5 to 100 mg and in other dosage forms, the content of the foregoing binding molecule is about 10 to 250 mg. Single and combination therapies. The binding molecules of the present disclosure can be used to treat diseases and disorders that are improved, inhibited, or alleviated by reducing IL-4 activity. These disorders include those characterized by abnormal or excessive expression of IL-4 or by abnormal host responses to IL-4 production. This disclosure encompasses combination therapies in which the anti-IL-4Rα binding molecule (such as an antibody or antibody fragment) is administered in combination with a second therapeutic agent. Co-administration and combination therapies are not limited to simultaneous administration, but include treatment regimens in which the anti-IL-4Rα binding molecule is administered at least once during a treatment course involving administration of at least one other therapeutic agent to the patient. The second therapeutic agent can be another IL-4 antagonist (such as another binding molecule) or a soluble cytokine receptor, an IgE antagonist, an anti-asthma drug (corticosteroid, non-steroidal agent, β-agonist, leukotriene antagonist, xanthine, fluticasone, salmeterol, albuterol) that can be delivered by inhalation or other suitable means. In a particular embodiment, the anti-IL-4Rα binding molecule (such as the binding molecules of this disclosure) can be administered with an IL-1 antagonist (such as rilonacept) or an IL-13 antagonist. In some aspects, the anti-IL-4Rα binding molecule (such as the binding molecules of this disclosure) can be administered in combination with binding molecules that target cytokines and / or receptors in type 1 or type 2 inflammatory responses. The second agent can include one or more leukotriene receptor antagonists for treating conditions such as allergic inflammatory conditions, such as asthma and allergies. Examples of leukotriene receptor antagonists include, but are not limited to, montelukast, pranlukast, and zafirlukast. The second agent can include cytokine inhibitors, such as one or more of the following: TNF (etanercept, ENBREL TM ), IL-9, IL-5, or IL-17 antagonists. Therapeutic Uses This disclosure provides compositions and methods for treating a disease or condition in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of a binding molecule of this disclosure. This disclosure provides compositions and methods for treating a disease or condition in an individual in need thereof, the method comprising administering to the individual a therapeutically effective amount of a composition of this disclosure. In certain embodiments of this disclosure, the disease or condition is a type 1 or type 2 inflammatory disease or condition. In certain embodiments of this disclosure, the disease or condition is an autoimmune disease or condition. In certain embodiments of this disclosure, the disease or condition is an allergic disease or condition. In certain embodiments of this disclosure, the disease or condition is an immune disease or condition. In certain embodiments of the present disclosure, the disease or disorder is a benign proliferative disease or disorder. In certain embodiments of the present disclosure, the disease or disorder is a malignant proliferative disease or disorder. In certain embodiments of the present disclosure, the disease or disorder is atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis, nasal polyps, or a combination thereof. In certain embodiments of the present disclosure, the binding molecule of the present invention is administered systemically. In one aspect, the binding molecule is administered intravenously or subcutaneously. In another aspect, the binding molecule is administered by injection or infusion. In another aspect, the binding molecule is administered by subcutaneous injection. In certain embodiments of the present disclosure, the binding molecule is administered systemically. In one aspect, the binding molecule is administered by subcutaneous injection. In another aspect, the therapeutically effective dose comprises a subcutaneous injection of about 75 mg, 150 mg, 300 mg, or 600 mg. In another aspect, the therapeutically effective dose comprises at least one subcutaneous injection, at least two subcutaneous injections, at least three subcutaneous injections, or at least four subcutaneous injections of about 75 mg, 150 mg, 300 mg, or 600 mg. In yet another aspect, the therapeutically effective dose comprises a subcutaneous injection of about 75 mg, 150 mg, 300 mg, or 600 mg once a week, once every two weeks, once every four weeks, or a maintenance dose is administered long-term to control disease symptoms. In certain embodiments of the present disclosure, the binding molecule is administered systemically. In one aspect, the binding molecule is administered by subcutaneous injection. In another aspect, the therapeutically effective dose comprises an initial dose of about 600 mg. In yet another aspect, the initial dose comprises an injection of 300 mg each in one of two different injection sites. In yet another aspect, including those in which the therapeutically effective dose comprises an initial dose, the therapeutically effective dose comprises a maintenance dose of about 300 mg. In another aspect, the maintenance dose is administered every other week. In certain embodiments of the method of the present disclosure, the binding molecule is administered systemically. In one aspect, the binding molecule is administered intravenously at a dose of about 1.0 mg / kg, 3.0 mg / kg, 8.0 mg / kg, or 12.0 mg / kg. In certain embodiments of the present disclosure, the binding molecule is administered in combination with a second therapeutic agent. In one aspect, the second therapeutic agent comprises an immunosuppressant. In one aspect, the second therapeutic agent comprises an agonist antibody. In one aspect, the second therapeutic agent comprises an immune activator. In another aspect, the second therapeutic agent comprises an IL-1β inhibitor, an IL-5 inhibitor, an IL-9 inhibitor, an IL-3 inhibitor, an IL-13 inhibitor, an IL-17 inhibitor, an IL-25 inhibitor, a TNFα inhibitor, an eotaxin-3 inhibitor, an IgE inhibitor, a prostaglandin D2 inhibitor, an immunosuppressant, a corticosteroid, a glucocorticoid, a proton pump inhibitor, a non-steroidal anti-inflammatory drug (NSAID), or a combination thereof. In certain embodiments of the present disclosure, the binding molecule is administered in combination with a second therapeutic agent. In one aspect, the second therapeutic agent comprises a corticosteroid. In a particular aspect, the corticosteroid is a topical corticosteroid. It should be understood that the compositions, formulations, kits, and methods of the present disclosure are not limited to any one disease and / or medical condition. The compositions, formulations, kits, and methods of the present disclosure can be applied to any disease and / or medical condition in which a patient exhibits an existing reactivity to a therapeutic antibody. Table 1: Exemplary sequences of the present disclosure. Examples Example 1: Substituted IgG4 C H The 3 constant regions reduce the existing immunoreactivity in some patient samples. Competitive inhibition studies were conducted to characterize the high existing reactivity signals observed in some samples. Commercially available antibody reagents or monoclonal antibodies specifically constructed for this purpose were used in these studies. A list of antibody constructs or reagents that help to elucidate the specificity of this existing reactivity is provided in Table 2. Table 2 The antibody constructs at 200 μg / mL were used as competitive inhibitors in the anti-drug antibody (ADA) confirmation assay format. The assay format is shown in Figure 4 and the assay results are shown in Figure 1 (Figure 1 and Figure 4). A high inhibition percentage in the assay indicates that the designated competitor is able to inhibit the reactive signal already present in these samples, suggesting that the competitor molecule contains a region of pre-existing reactive binding. A lower inhibition percentage indicates that the competitor molecule does not contain a region of pre-existing reactive binding. The reagents listed in Table 1 above can be broadly classified into three categories, IgG4 backbone / Fc hinge region constructs, commercially acquired isotype antibodies, and bioengineered constructs of dupilumab. Preliminary experiments examined whether this high assay reactivity was directed against the CDR of dupilumab or the IgG4 backbone. REGN-A is a human monoclonal antibody with the same IgG4 backbone as dupilumab but different CDR sequences and does not bind to IL-4Rα. As shown in Figure 1, a competitive inhibition experiment using both dupilumab and REGN-A demonstrated significant inhibition of the high assay signal in 6 baseline samples from the patients examined. Since the CDR region sequences of these two antibodies are different, this inhibition result indicates that the high signal reactivity is not directed against the CDR portion of dupilumab, but rather against some common antibody backbone sequences. Both dupilumab and REGN-A contain the IgG1 "CPPC" hinge region sequence that stabilizes the antibody hinge region in these IgG4 molecules. To determine whether the pre-existing immunoreactivity was targeted to this CPPC mutation, a commercially available IgG4κ antibody with a wild-type (wt) CPSC sequence in the hinge region was examined in the competitive inhibition experiment. As can be seen from Figure 1, this wild-type IgG4 antibody also showed significant inhibition similar to that of dupilumab and REGN-A. This result indicates that the high baseline assay signal is not directed against the CPPC hinge region mutation in dupilumab, but most likely against the wild-type constant region of the IgG4 molecule. These results indicate that the high signal observed at baseline is not specific to dupilumab. Additional experiments were conducted to determine whether this high assay pre-existing reactivity was directed against constant region sequences common between the different IgG subtypes. Using the same competitive inhibition method, the effects of three commercially acquired human IgG1κ, IgG2κ, IgG3κ antibodies on this pre-existing reactivity were examined. None of these antibodies were able to inhibit the high baseline signal in these samples (see Figure 1). This suggests that the pre-existing reactivity is most likely related to a region specific to the IgG4 constant region sequence and not shared by any of the IgG subtypes tested. Constructed similar to dupilumab, but with IgG4 C H domain 3 replaced by IgG1 C H3 - domain substituted human monoclonal antibody (REGN - B) (see Figure 2). This antibody was examined in competition studies to determine whether the existing reactivity targets the C H 3 - domain. REGN - B did not significantly inhibit the high signal in the samples (see Figure 1), indicating that the existing reactivity most likely targets some regions within the C H 3 - domain of the IgG4 molecule. To further identify the regions that might be associated with these high signals within the C H 3 - domain of dupilumab, amino acid sequence alignments of the C H 3 - domains of IgG4, IgG1, and IgG2 antibodies were performed (see Figure 3). It was noted that there were differences at six individual amino acid positions between the IgG4 and IgG1 C H 3 - domain sequences, and differences at five individual amino acid positions between the IgG4 and IgG2 C H 3 - domain sequences (see Figure 3). An IgG4 construct with leucine (L) substituted by proline (P) at position 445 was obtained and examined in a competitive inhibition assay. This construct did not show significant inhibition of these high assay signals. This indicates that the existing reactivity might be specific to the L445 region of dupilumab. Another sample with a high assay signal was examined using this construct with L substituted by P at position 445 in the competitive inhibition assay and a similar low level of inhibition was observed. This seems to confirm that the existing reactivity is specifically targeted to the region around L445. Thus, the change from leucine (in wt IgG4) to proline (present at the same position in wt IgG1, IgG2, and IgG3) in dupilumab eliminates the existing reactivity that leads to high signals in the ADA assay. An IgG4 construct with leucine (L) substituted by proline (P) at position 445 was obtained and examined in a competitive inhibition assay. This construct did not indicate significant inhibition of these high assay signals. This suggests that the existing reactivity may be specific to the L445 region of dupilumab. Another sample with a high assay signal was examined using this construct with L substituted by P at position 445 in the competitive inhibition assay and a similar low level of inhibition was observed. This seems to confirm that the existing reactivity specifically targets the region around L445. Thus, the change in dupilumab from leucine (in wt IgG4) to proline (the same position as present in wt IgG1, IgG2, and IgG3) eliminates the existing reactivity that results in high signals in the ADA assay. A second dupilumab (REGN-C) based on a human monoclonal antibody construct was generated. This construct is identical to dupilumab except for a point mutation at residue 445 in the antibody sequence where leucine was changed to proline (abbreviated as L>P). REGN-C could not significantly inhibit the high signal in the baseline sample of the test (see Figures 1 and 2). This confirms that the existing reactivity specifically targets the region around L445 in dupilumab and suggests that using this engineered dupilumab in the ADA assay eliminates most (if not all) of the high-level background signals observed in the current ADA assay. Further experiments were conducted using two additional antibodies, REGN-F and REGN-G, to demonstrate that the modification of residue 445 in the antibody sequence from leucine to proline eliminated the pre-existing reactivity. As shown in Figure 7, competitive inhibition experiments using dupilumab, REGN-F, and REGN-G showed significant inhibition of the high assay signal in the six baseline samples, thus indicating that dupilumab, REGN-F, and REGN-G exhibited a high degree of pre-existing reactivity. However, substitution of leucine (L) with proline (P) at position 445 in REGN-F and REGN-G (designated herein as REGN-F(L445P) and REGN-G(L445P), respectively) eliminated this pre-existing reactivity, as neither REGN-F nor REGN-G could inhibit the high assay signal. REGN-F, REGN-F(L445P), REGN-G, and REGN-G(L445P) were also tested as reagents in a drug-specific bridging anti-drug antibody assay (similar to that described in Figure 4). As shown in Figures 8 and 9, high assay signals indicative of a high degree of pre-existing reactivity were demonstrated when both REGN-F and REGN-G were used as capture and detection agents. In contrast, the assay signal was significantly reduced when REGN-F(L445P) was used as the capture agent in combination with REGN-F as the detection agent (Figure 8), or when REGN-G was used as the capture agent in combination with REGN-G(L445P) as the detection agent (Figure 9), demonstrating that the L445P substitution eliminated the pre-existing reactivity. REGN-F and REGN-G contain different variable domains that are also different from the variable domains of the antibodies tested in the results shown in Figure 1. Thus, the results shown in Figures 7 to 9 demonstrate that the pre-existing reactivity is not related to the variable domain and CDR, but is specific to the region around L445. In addition, the L445P substitution was shown to be generally applicable to IgG4 antibodies to reduce pre-existing reactivity regardless of the nature of their CDRs. At least most of the regions of pre-existing reactivity in the dupilumab antibody sequence have been identified. The high signal in the ADA assay appears to be due to some matrix components in these serum samples, which can bridge the labeled dupilumab molecules in the assay by binding at or near L445 in the C H 3 domain. This indicates that the pre-existing reactivity is not specific to the dupilumab drug, but can bind to any IgG4 molecule. In addition, the results suggest that using a modified version of dupilumab with the L445P mutation in the ADA assay eliminates most, if not all, of the high background signal. Example 2: Development of a modified anti-drug antibody (ADA) assay to reduce background immunoreactivity in patient samples Develop a modified ADA assay using biotinylated REGN-C (with the L445P mutation) as a capture agent. Figure 4 illustrates the differences in the assay design between the current ADA assay and the modified ADA assay using REGN-C. For clarity, the current ADA assay will be referred to as "ADA assay #1", and the modified ADA assay will be called "ADA assay #2". Figure 5 shows a comparative analysis of the ADA screening signals obtained from all patient baseline samples using the current ADA assay #1 versus ADA assay #2. Figure 5A shows a plot of the signal-to-noise ratio generated by these high ADA signal baseline samples in the current ADA screening assay (assay #1), and Figure 5B shows a plot of the signal-to-noise ratio generated by the exact same samples in ADA assay #2. The assay format of ADA assay #2 significantly reduces the high signals observed in the current ADA screening assay. Some samples in ADA assay #2 still demonstrate reactivity, but the number of screening positives decreases more, consistent with the expected false positive rate of the screening assay, and the signal response of those positive samples is generally much lower than that observed using the current ADA assay. In ADA assay #2, the observation that in most cases these high assay signals decline to near baseline values should allow for improved detection of treatment-emergent and drug-specific ADA in the patient population. Incorporated by reference Unless expressly excluded or otherwise limited, the entire content of each document cited herein, including any cross-referenced or related patent or application, is incorporated herein by reference. The citation of any document is not an admission that it is prior art with respect to any disclosure herein or that it alone or in any combination with any other reference teaches, suggests, or discloses any such disclosure. Moreover, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to the term in this document shall control. Other embodiments Although certain embodiments of the present disclosure have been illustrated and described, various other changes and modifications can be made without departing from the spirit and scope of the present disclosure. The appended claims cover all such changes and modifications that are within the scope of the present disclosure. [Figure 1] is a schematic diagram depicting the specificity of the reactivity already present in the baseline patient samples when using competitive antibody constructs in anti-drug antibody (ADA) assays. The percentage of inhibition on the y-axis ranges from 0 to 100, with an increment of 25. On the x-axis are the specific competitive antibody constructs used in the bridging assay, arranged from left to right as follows: dupilumab, REGN-A, wt IgG4, wt IgG1, wt IgG2, wt IgG3, REGN-B, and REGN-C. The samples (S1 - S6) from the six patients are presented in sequence as a circle, a square, an upward-pointing triangle, a star, a diamond, and a downward-pointing triangle. The antibody reagents mentioned in the figure are used as competitive inhibitors in the ADA confirmation assay format at a dosage of 200 μg / mL. A high percentage of inhibition in the assay indicates that the designated competitor is able to inhibit the signal already present in these samples, suggesting that the competitor molecule contains a region that binds to the existing reactivity. A lower percentage of inhibition indicates that the competitor molecule does not contain a region that can bind to the existing reactivity. [Figure 2] is a graph showing the schematic representation of the three key constructs and their structural differences compared to dupilumab. REGN-B, REGN-C, and REGN-E are molecular analogs of dupilumab. All three constructs are IgG4k constructs, which have the same specificity and similar affinity for anti-IL14R as dupilumab, and also possess the same CPPC hinge region mutation sequence present in dupilumab. [Figure 3] is a graph showing the schematic representation of the amino acid sequence alignment and the comparison of the C H 3 domains. The leucine at position 445 is the third from the C-terminus of the C H 3 domain sequence, as indicated by the arrow. [Figure 4] is an embodiment that illustrates the schematic representation of drug-specific bridging (Figure A, ADA assay #1), non-drug-specific bridging due to the reactivity already present in the current ADA assay (Figure B), and non-drug-specific bridging due to the use of REGN-C as a capture agent (Figure C, ADA assay #2). Streptavidin on the plate is shown as a gray cross, the biotin moiety is shown as a small black square, and the ruthenium label is shown as a star. The biotinylated REGN-D (dupilumab) capture agent (black fork structure) is shown in Figures A and B, while biotinylated REGN-C is shown in Figure C (black and white square fork structure). The existing reactivity is shown as a smaller dashed-outline bifurcated structure in Figures B and C. [Figure 5A] is a schematic diagram showing the analysis signal from a subset of patient baseline samples, and the analysis signal is obtained using the original ADA analysis (Analysis #1) (Figure A in Figure 4 is its illustration and uses biotinylated REGN-D as the capture agent). The signal / noise ratio is represented on the y-axis on a logarithmic scale from 0 to 100. The individual patient samples sorted by the signal / noise ratio are on the x-axis. The cut-off point is indicated by a dashed line. [Figure 5B] is a schematic diagram showing the analysis signal of the baseline sample of the same patient, and the analysis signal is obtained using the modified ADA analysis (Analysis #2) (Figure C in Figure 4 is its illustration and uses biotinylated REGN-C as the capture agent). The signal / noise ratio is represented on the y-axis on a logarithmic scale from 0 to 100. The order of the individual patient samples on the x-axis is the same as that shown in Figure 5A. The cut-off point is indicated by a dashed line. [Figure 6A] is a schematic diagram showing the analysis signal from a subset of patient baseline samples, and the analysis signal is obtained using a drug-specific bridging anti-drug antibody (ADA) analysis (similar to that described in Figure 4) and using intact dupilumab as the capture agent and the detection agent. [Figure 6B] is a schematic diagram showing the analysis signal from the same subset of patient baseline samples tested in Figure 6A, and the analysis signal is obtained using the modified bridging ADA analysis and using REGN-B (Figure 2, a human IgG4 mAb, where the entire C H 3 of IgG4 is converted to IgG1 C H 3 domain) as the capture agent and the detection agent. [Figure 6C] is a schematic diagram showing the analysis signal from the same subset of patient baseline samples tested in Figure 6A, and the analysis signal is obtained using the modified bridging ADA analysis and using REGN-E (Figure 2, a human IgG4 mAb, where the C H 3 domain is truncated by a stop codon after serine 444) as the capture and detection agent. [Figure 7] is a series of schematic diagrams characterizing the specificity of pre - existing reactivity in a baseline patient sample in an anti - drug antibody (ADA) assay using competitive antibody constructs. The percent inhibition on the y - axis ranges from 0 to 100 in increments of 25. On the x - axis are the specific competitive antibody constructs used in the bridging assay, listed from left to right as: upper schematic: dupilumab, REGN - F, and REGN - F (L445P), or lower schematic: dupilumab, REGN - G, and REGN - G (L445P). The six patient samples (S1 - S6) are presented in order as circles, squares, upward triangles, stars, diamonds, and downward triangles. The antibody reagents mentioned in the figure are used as competitive inhibitors in the ADA confirmation assay format at a dose of 200 μg / mL. A high percent inhibition in the assay indicates that the designated competitor can inhibit the pre - existing signal in these samples, suggesting that the competitor molecule contains a region of pre - existing reactive binding. A lower percent inhibition indicates that the competitor molecule does not contain a region of pre - existing reactive binding. [Figure 8] is a series of schematic diagrams showing the assay signals from a subset of patient baseline samples, which were assayed using a drug - specific bridging anti - drug antibody (ADA) assay (similar to that described in Figure 4) and using REGN - F as both the capture and detection agent (upper), or using REGN - F (L445P) as the capture agent in combination with REGN - F as the detection agent. [Figure 9] is a series of schematic diagrams showing the assay signals from a subset of patient baseline samples, which were assayed using a drug - specific bridging anti - drug antibody (ADA) assay (similar to that described in Figure 4) and using REGN - G as both the capture and detection agent (upper), or using REGN - G as the capture agent in combination with REGN - G (L445P) as the detection agent.

Claims

1. A non-naturally produced monoclonal antibody comprising a C-terminal heavy chain sequence comprising a sequence selected from the group consisting of: SEQ ID NO: 7 and SEQ ID NO:

13.

2. The antibody as claimed in claim 1, wherein the C-terminal heavy chain sequence contains SEQ ID NO:

7.

3. The antibody as claimed in claim 1, wherein the C-terminal heavy chain sequence contains SEQ ID NO:

13.

4. The antibody as claimed in claim 1, wherein the C-terminal heavy chain sequence comprises: a CH domain sequence comprising SEQ ID NO:

8.

5. The antibody as claimed in claim 1, wherein the C-terminal heavy chain sequence comprises: a CH domain sequence comprising SEQ ID NO:

22.

6. The antibody of claim 1, wherein the antibody comprises: the amino acid sequence of the heavy chain variable region (VH) complementarity-determining region (CDR) CDR1 of SEQ ID NO: 9; the amino acid sequence of VH CDR2 of SEQ ID NO: 10; the amino acid sequence of VH CDR3 of SEQ ID NO: 11; the amino acid sequence of the light chain variable region (VL) CDR1 of SEQ ID NO: 12; the amino acid sequence of VL CDR2 of LGS; and the amino acid sequence of VL CDR3 of SEQ ID NO:

14.

7. The antibody of claim 1, wherein the antibody comprises VH and VL, the VH comprising the amino acid sequence of SEQ ID NO: 15, and the VL comprising the amino acid sequence of SEQ ID NO:

16.

8. The antibody of claim 1, wherein the antibody comprises an IgG4 CH1 region comprising the amino acid sequence of SEQ ID NO:

1.

9. The antibody of claim 1, wherein the antibody comprises an IgG4 CH2 region comprising the amino acid sequence of SEQ ID NO:

2.

10. An analysis comprising (a) a solid support, wherein a first component is operatively connected to the solid support; (b) at least one capture agent, wherein a second component is operatively connected to the at least one capture agent; wherein the capture agent comprises a first non-naturally generated monoclonal antibody as claimed in any one of claims 1 to 9; and (c) at least one detection agent, wherein a detectable label is operatively connected to the detection agent, wherein the detection agent comprises a second non-naturally generated monoclonal antibody as claimed in any one of claims 1 to 9, and wherein the first component and the second component selectively bind to each other.

11. As requested in item 10, wherein the assay contains dupilumab.

12. As described in claim 10, wherein the first component contains streptavidin and the second component contains biotin.

13. An analysis comprising (a) a solid support, wherein a first component is operatively connected to the solid support; (b) at least one capture agent, wherein a second component is operatively connected to the at least one capture agent and wherein the capture agent comprises a non-naturally generated monoclonal antibody as claimed in any one of claims 1 to 9; and (c) at least one detection agent, wherein a detectable label is operatively connected to the detection agent and wherein the detection agent comprises dupilumab; wherein the first component and the second component selectively bind to each other.

14. As described in claim 13, wherein the first component contains streptavidin and the second component contains biotin.

15. As claimed in claim 13, wherein i) the at least one capture agent does not bind to an antibody that does not specifically bind to the variable region sequence of dupilumab; and / or ii) the at least one capture agent and the at least one detection agent bind to an antibody that specifically binds to the variable region sequence of dupilumab.

16. A method for determining the degree of immunogenicity of a monoclonal antibody therapy in an individual, comprising (a) contacting a biological sample from the individual with an analysis as claimed in claim 13 under conditions suitable for allowing at least one antibody in the biological sample to bind with at least one capture agent and at least one detection agent, wherein the individual has been administered the monoclonal antibody therapy prior to the contact step, (b) detecting a signal from the at least one detection agent, and (c) identifying the degree of immunogenicity of the individual as high when the signal from (b) is above a threshold, or (d) identifying the degree of immunogenicity of the individual as low when the signal from (b) is below a threshold.

17. The method of claim 16, wherein the monoclonal antibody therapy comprises an antibody that includes a C-terminal heavy chain sequence comprising a sequence selected from the group consisting of: SEQ ID NO: 7 and SEQ ID NO:

13.

18. The method of request item 16, wherein the threshold is a predetermined value or a safety threshold.

19. The method of claim 16, wherein the monoclonal antibody therapy is i) initiating and the immunogenicity level is at a baseline level; ii) ongoing and the immunogenicity level is at a subsequent level; and iii) ending and the immunogenicity level is at a final level.

20. The method of claim 16, wherein the individual suffers from an inflammatory disease or condition, an allergic disease or condition, an immune disease or condition, a benign proliferative disease or condition, or any combination thereof.

21. The method of claim 20, wherein the individual suffers from an autoimmune disease or condition, atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis, nasal polyps, or any combination thereof.

22. The method of claim 16, wherein the monoclonal antibody therapy comprises dupilumab, wherein the biological sample comprises an antidrug antibody (ADA) against dupilumab.

23. The method of claim 20, wherein the biological sample comprises blood, serum or plasma.

24. A method for determining the degree of immunogenicity of a monoclonal antibody therapy in an individual, comprising (a) contacting a biological sample from the individual with an analytical contact comprising (i) a solid support comprising streptavidin operably linked to the solid support; (ii) at least one capture agent comprising biotin operably linked to an isolated monoclonal antibody comprising a C-terminal heavy chain sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 7 and SEQ ID NO: 13; and (iii) at least one detection agent comprising a detectable labeled dupilumab; wherein the streptavidin selectively binds to the biotin to bind the at least one capture agent to the support, and at least one antibody in the biological sample binds the at least one capture agent to the at least one detection agent, and wherein the individual has been administered the monoclonal antibody therapy prior to the contact step. (b) Detect a detectable signal from the at least one detection agent, and (c) when the signal from (b) is above a threshold, identify the degree of immunogenicity of the monoclonal antibody therapy in the individual as high, or (d) when the signal from (b) is below the threshold, identify the degree of immunogenicity of the monoclonal antibody therapy in the individual as low.

25. The method of claim 24, wherein the monoclonal antibody therapy comprises an antibody comprising a C-terminal heavy chain sequence comprising a sequence selected from the group consisting of SEQ ID NO: 7 and SEQ ID NO:

13.

26. The method of request item 24, wherein the threshold is a predetermined value or a safety threshold.

27. The method of claim 24, wherein the monoclonal antibody therapy is: i) initiating and the immunogenicity level is at baseline; ii) ongoing and the immunogenicity level is at subsequent level; or iii) ending and the immunogenicity level is at final level.

28. The method of claim 24, wherein the individual suffers from an inflammatory disease or condition, an autoimmune disease or condition, an allergic disease or condition, an immune disease or condition, or a benign proliferative disease or condition.

29. The method of claim 24, wherein the individual is a patient receiving medical treatment.

30. The method of claim 24, wherein the individual is a participant in a clinical trial.

31. The method of claim 24, wherein the monoclonal antibody therapy comprises dupilumab.

32. The method of claim 24, wherein the individual has been given a therapeutically effective dose of the monoclonal antibody therapy.

33. The method of claim 24, wherein the detectable marker is selected from the group consisting of: ruthenium, horseradish peroxidase, alkaline phosphatase and luciferase.

34. The method of claim 24, wherein the individual suffers from atopic dermatitis, asthma, allergic rhinitis, allergic conjunctivitis, eosinophilic esophagitis, nasal polyps, or any combination thereof.

35. The method of claim 24, wherein the at least one antibody in the biological sample is an anti-drug antibody (ADA).

Images