Anti-il-18BP antibodies

Abstract

Antibodies that bind to interleukin-18 binding protein (IL-18BP), and related compositions, are provided that can be used in any of a variety of therapeutic or diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 437,526, filed on January 6, 2023, U.S. Provisional Patent Application No. 63 / 590,348, filed on October 13, 2023, and U.S. Provisional Patent Application No. 63 / 596,580, filed on November 6, 2023, the contents of which are incorporated herein by reference in their entirety. Background Art

[0003] Interleukin-18 (IL-18) is an immunostimulatory cytokine with anti-tumor activity. It plays a key role in linking inflammatory immune responses to tumor progression. Although recombinant human IL-18 has been evaluated as a cancer immunotherapy, this approach is ineffective due, at least in part, to a feedback loop in humans, in which administration of IL-18 leads to increased production of IL-18BP, which neutralizes the administered IL-18 cytokine (see, e.g., Robertson et al., Clinical Cancer Res. 12:4265-4273, 2006).

[0004] IL-18BP is a high-affinity IL-18 decoy receptor that is frequently raised in tumors. Studies have shown that IL-18BP is a secretory immune checkpoint and an obstacle to IL-18 immunotherapy (see, for example, Zhou et al., Nature (Nature) 583 (7817): 609-614, 2020). It is believed that IL-18BP inhibits the pro-inflammatory activity of IL-18 by sequestering IL-18 away from cell surface receptors. The affinity of IL-18 for IL-18BP is higher than the affinity of IL-18 for IL-18 receptors, and IL-18BP is often present in an amount exceeding IL-18, thereby ensuring strict regulation. It has also been shown that IL-18BP especially balances Th1 and Th2 immune responses and plays a key role in autoimmune diseases (see, for example, Park et al., Biomedicines (Biomedicines) 10 (7): 1750, 2022). Therefore, there is a need for agents and methods that specifically modulate the activity of IL-18BP, and such agents and methods are provided herein. Summary of the Invention

[0005] The present invention relates to antibodies and related compositions that bind to interleukin-18 binding protein (IL-18BP) and can be used in any of a variety of therapeutic and diagnostic methods, including the treatment or diagnosis of cancer and other diseases.

[0006] Aspects of the present invention include an antibody that binds to interleukin-18 binding protein (IL-18BP), wherein at least one antibody competes with IL-18 for binding to IL-18BP. In another aspect, the present invention provides an antibody that binds to IL-18BP and interferes with the binding of IL-18 to IL-18BP. In another aspect, the present invention provides an antibody that binds to IL-18BP and is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18. In some embodiments, the present invention provides an antibody that binds to a preformed IL-18-IL-18BP complex. In some embodiments, the present invention provides an antibody that binds to free IL-18BP. In some embodiments, the antibody binds to a conformational epitope of IL-18BP. In some embodiments, the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In some embodiments, the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In some embodiments, the antibody binds to a linear epitope of IL-18BP. In some embodiments, the antibody binds to the binding interface between IL-18 and the mature form of IL-18BP. In some embodiments, the antibody binds to amino acid residues S75, H79, T116, and S119 of SEQ ID NO: 372.

[0007] In some embodiments, the antibody binds to human IL-18BP and cynomolgus monkey IL-18BP but not to mouse IL-18BP. In some embodiments, the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

[0008] In some embodiments, the present invention further comprises an antibody that binds to interleukin-18 binding protein (IL-18BP), wherein at least one antibody comprises: a heavy chain variable region (V H ), which comprises a complementarity determining region V selected from Table A1 H CDR1, V H CDR2 and V H CDR3 sequences and variants thereof that specifically bind to IL-18BP; and light chain variable region (V L ), which comprises a complementarity determining region V selected from Table A1 L CDR1, V L CDR2 and V LCDR3 sequences and their variants that specifically bind to IL-18BP.

[0009] In some embodiments, the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein said V H In some embodiments, the V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein said V L There are at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in the framework regions.

[0010] Also included are isolated polynucleotides encoding the anti-IL-18BP antibodies described herein; expression vectors comprising the isolated polynucleotides; and isolated host cells comprising the vectors. Also provided are one or more isolated polynucleotides encoding the anti-IL-18BP antibodies described herein. For example, provided herein is a first polynucleotide encoding a V or V sequence of an antibody disclosed herein. H region; and a second polynucleotide encoding the V L district.

[0011] Certain embodiments include a pharmaceutical composition comprising an anti-IL-18BP antibody described herein and a pharmaceutically acceptable carrier. In some embodiments, the composition is a sterile injectable solution optionally suitable for intravenous, intramuscular, subcutaneous, or intraperitoneal administration.

[0012] Also included are methods of treating a disease or condition in a subject in need thereof, comprising administering to the subject a pharmaceutical composition as described herein. In some embodiments, the disease or condition is a cancer or tumor or a proliferative disease or condition, optionally selected from the group consisting of a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enteritis, a proliferative diabetic retinopathy, and a proliferative nephropathy. In some embodiments, the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or the proliferative disease or condition is associated with increased expression of IL-18BP and / or IL-18. In some embodiments, the cancer is selected from one or more of the following: bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma), 'slymphoma), Hodgkin's lymphoma), liver cancer (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, urothelial cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and stomach cancer.

[0013] Also included are methods for screening anti-IL-18BP antibodies for the ability to block or inhibit binding between IL-18 and IL-18BP, comprising (a) determining the binding affinity of the antibody to (i) IL-18BP alone and (ii) a low IL-18 fusion protein, wherein the low IL-18 fusion protein comprises IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein binds to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein; (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) if the binding affinity of (i) is significantly stronger than the binding affinity of (ii), identifying or selecting the antibody as capable of blocking or inhibiting binding between IL-18 and IL-18BP.

[0014] In some aspects, the present invention includes an isolated low IL-18 fusion protein comprising, in an N-terminal to C-terminal orientation, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and an IL-18BP. In some embodiments, the low IL-18 fusion protein comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to a sequence from Table S1.

[0015] Also included are methods of stimulating an immune response in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein. In some embodiments, the immune response is an IL-18-mediated immune response. In specific embodiments, the IL-18-mediated immune response comprises inducing IFN-γ, CXCL10, and / or TNFα in the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1A-Figure 1C The repertoire and low IL-18 design of the antibodies of the invention are shown. Figure 1A The repertoire of the antibodies of the invention is shown. Figure 1B A schematic diagram showing an exemplary low IL-18 expression cassette. The signature of the gene from N' to C' terminus includes: the mycosin signal peptide, the human IL-18 coding region, a flexible gly-ser linker interspersed with a tobacco etch virus (TEV) protease cleavage site, the human IL-18BP coding sequence, and a 6x HIS tag. Figure 1C A view showing a crystal structure-derived model of human IL-18 in complex with Ectromeliavirus IL-18BP (as indicated). The IL-18BP N-terminus and IL-18 C-terminus are indicated by boxes and arrows.

[0017] Figure 2A-2G The effect of anti-IL-18BP antibody on IL-18-mediated induction of IFNγ in healthy human peripheral blood mononuclear cells (PBMC) in vitro is shown. Anti-IL-18BP antibody exhibits a dose-dependent increase in IFNγ.

[0018] Figure 3A-3G Figure 3 depicts the ability of anti-IL-18BP antibodies to release IL-18 from pre-existing complexes with IL-18BP. Anti-IL-18BP antibodies were added to human PBMCs after IL-18 was allowed to form a pre-complex with IL-18BP, which exhibited a dose-dependent increase in IFNγ.

[0019] Figure 4The ability of anti-human IL-18BP antibodies to elicit IL-18-induced cytokine CCL2 production in human PBMCs as measured by ELISA is depicted.

[0020] Figure 5A-5B Results of cynomolgus macaque PBMC analysis are shown. Figure 5A The effect of anti-IL-18BP antibody on IL-18-mediated in vitro induction of IFNγ in cynomolgus macaque PBMCs is shown. Figure 5B The presentation depicts the ability of anti-IL-18BP antibodies to release IL-18 from a pre-existing complex with IL-18BP via IFNγ release.

[0021] Figure 6 Depicted is the ability of IL-18BP antibodies to disrupt the IL-18:IL-18BP complex and induce IFNγ secretion from NK cells. DETAILED DESCRIPTION

[0022] The present invention relates to antibodies that specifically bind to interleukin-18 binding protein (IL-18BP). Some embodiments include specific humanized antibodies that are capable of binding to IL-18BP, blocking or reducing the inhibitory binding of IL-18BP to its ligand IL-18, and thereby increasing IL-18-mediated downstream signaling. Therefore, in certain embodiments, the anti-IL-18BP antibody is an IL-18BP antagonist or inhibitor.

[0023] The IL-18BP antagonist antibodies described herein are useful for treating and preventing a variety of diseases and conditions, such as cancer and other diseases and conditions. Some embodiments therefore relate to the use of anti-IL-18BP antibodies for diagnosing, assessing, and treating diseases and conditions, including those associated with IL-18 and / or IL-18BP activity or aberrant expression thereof.

[0024] Unless specifically indicated to the contrary, the practice of the present invention employs conventional methods of virology, immunology, microbiology, molecular biology and recombinant DNA technology within the skill of the art, many of which are described below for illustrative purposes. Such techniques are fully explained in the literature. See, e.g., Current Protocols in Molecular Biology or Current Protocols in Immunology, John Wiley & Sons, New York, NY (2009); Ausubel et al., Short Protocols in Molecular Biology, 3rd ed., Wiley & Sons, 1995; Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd ed., 2001); Maniatis et al., Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, Volumes I and II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Nucleic Acid Hybridization. Hybridization (B. Hames and S. Higgins, eds., 1985); Transcription and Translation (B. Hames and S. Higgins, eds., 1984); Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984), and other similar references.

[0025] definition

[0026] Unless otherwise defined, all technical terms, notation, and other technical and scientific terms or nouns used herein are intended to have the same meanings as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. In some cases, terms with commonly understood meanings are defined herein for clarity and / or ease of reference, and the inclusion of such definitions herein should not necessarily be construed to indicate a substantial difference from what is generally understood in the art.

[0027] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0028] As used herein, the term "about" will be understood by one of ordinary skill in the art and will vary to some extent depending on the context in which it is used. In some embodiments, the term "about," when referring to a measurable value such as an amount, duration, etc., is intended to encompass art-accepted variations based on the standard error in making such measurements. In some embodiments, the term "about," when referring to such a value, is intended to encompass variations of ±10% from the specified value.

[0029] As used herein, the term "antibody" encompasses not only intact polyclonal or monoclonal antibodies, but also antigen-binding fragments thereof (such as dAb, Fab, Fab', F(ab')2, Fv), single chains (scFv), synthetic variants thereof, naturally occurring variants, fusion proteins comprising an antibody portion having an antigen-binding fragment with the desired specificity, humanized antibodies, chimeric antibodies, and any other modified configuration of an immunoglobulin molecule comprising an antigen-binding site or fragment (epitope recognition site) with the desired specificity. Certain features and properties of antibodies (and antigen-binding fragments thereof) are described in more detail herein.

[0030] As used herein, the term "antigen-binding fragment" refers to a polypeptide fragment that contains at least one CDR of an immunoglobulin heavy chain and / or light chain and binds to an antigen of interest. In this regard, an antigen-binding fragment of an antibody described herein may comprise a V or V region from an antibody that binds to a target molecule. H and V L In certain embodiments, the antigen-binding fragments of the invention comprise 1, 2, 3, 4, 5 or all 6 CDRs of the sequences disclosed herein. H and V L All 6 CDRs of the sequence.

[0031] The binding properties of antibodies and their antigen-binding fragments can be quantified using methods well known in the art (see Davies et al., Annual Review of Biochemistry (Annual Rev. Biochem.) 59:439-473, 1990). In certain embodiments, antibodies as described herein include heavy chain and light chain complementary determining regions (CDR) sets inserted between heavy and light chain framework regions (FR) sets, which provide support for CDRs and define the spatial relationship of CDRs relative to each other. As used herein, the term "CDR set" refers to three hypervariable regions in the heavy or light chain V region. Starting from the N-terminus of the heavy or light chain, these regions are respectively represented as "CDR1," "CDR2," and "CDR3." Thus, an antigen binding site includes six CDRs, comprising a CDR set from each of the heavy and light chain V regions. A polypeptide comprising a single CDR (e.g., CDR1, CDR2, or CDR3) is referred to herein as a "molecular recognition unit." Crystallographic analysis of various antigen-antibody complexes has shown that the amino acid residues of the CDRs form extensive contacts with the bound antigen, with the most extensive antigen contact being with heavy chain CDR3. Therefore, the molecular recognition unit is primarily responsible for the specificity of the antigen-binding site.

[0032] The structure and location of immunoglobulin variable domains can be determined by reference to Kabat, EA et al., Sequences of Proteins of Immunological Interest. 4th ed. US Department of Health and Human Services 1987 and its updated versions.

[0033] Also included are "monoclonal" antibodies, which refer to a homogeneous antibody population, wherein the monoclonal antibody comprises amino acids (naturally occurring amino acids and non-naturally occurring amino acids) that participate in the selective binding of the epitope. The term "monoclonal antibody" encompasses not only complete monoclonal antibodies and full-length monoclonal antibodies, but also fragments thereof (such as Fab, Fab', F(ab')2, Fv), single chains (ScFv), variants thereof, fusion proteins comprising antigen-binding portions, humanized monoclonal antibodies, chimeric monoclonal antibodies, and any other modified configurations of immunoglobulin molecules comprising antigen-binding fragments (epitope recognition sites) having the desired specificity and capable of binding to epitopes. It is not desired to limit the source of the antibody or its preparation method (e.g., by fusion tumors, phage selection, recombinant expression, transgenic animals). The term includes whole immunoglobulins and the fragments described above under the definition of "antibody," etc.

[0034] In certain embodiments, antibodies are made human-like by, for example, producing chimeric antibodies. Chimeric antibodies are generally prepared using recombinant technology, with an antigen binding site derived from an immunoglobulin from a non-human species and the remaining immunoglobulin structure of the molecule based on the structure and / or sequence of a human immunoglobulin. The antigen binding site may comprise a complete variable domain fused to a constant domain or only comprise CDRs (all or part) on an appropriate framework region transplanted into the variable domain. The epitope binding site may be wild type or modified by one or more amino acid substitutions. This eliminates the constant region as an immunogen in a human individual, but retains the possibility of an immune response to the foreign variable region (LoBuglio et al., PNAS USA 86:4220-4224, 1989; Queen et al., PNAS USA.86:10029-10033, 1988; Riechmann et al., Nature 332:323-327, 1988). Illustrative methods for antibody humanization include those described in U.S. Patent No. 7,462,697.

[0035] Another approach focuses not only on providing constant regions of human origin, but also on modifying the variable regions, thereby reshaping them into as close a human form as possible. It is known that the variable regions of the heavy and light chains contain three complementary determining regions (CDRs), which vary in response to the relevant epitopes and determine the binding capacity, flanked by four framework regions (FRs) that are relatively conserved in a given species and are assumed to provide a skeleton for the CDRs. When a non-human antibody is prepared for a specific epitope, the variable region can be "reshaped" or "humanized" by transplanting the CDRs derived from the non-human antibody onto the FRs present in the human antibody to be modified. The application of this method to various antibodies has been reported in Sato et al., Cancer Res. 53:851-856, 1993; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen et al., Science 239:1534-1536, 1988; Kettleborough et al., Protein Engineering 4:773-3783, 1991; Maeda et al., Human Antibodies Hybridoma 2:124-134, 1991; Gorman et al., PNAS USA 88:4181-4185, 1991; Tempest et al., Bio / Technology 9:266-271, 1991; Co et al., PNAS USA.88:2869-2873, 1991; Carter et al., PNAS USA.89:4285-4289, 1992; and Co et al., Journal of Immunology (J Immunol.) 148:1149-1154, 1992. In some embodiments, the humanized antibody retains all CDR sequences (e.g., a humanized mouse antibody containing all six CDRs from a mouse antibody). In some embodiments, only some CDR sequences are transplanted from non-human antibodies (Bowers et al., Journal of Biological Chemistry (J.Biol.Chem.) 288:7688-7696, 2013). In certain embodiments, the humanized antibody has one or more CDRs (one, two, three, four, five, six) that are changed relative to the original antibody, which is also referred to as one or more CDRs "derived from" one or more CDRs from the original antibody.

[0036] In certain embodiments, the antibody is a "chimeric" antibody. In this regard, a chimeric antibody comprises an antigen-binding fragment of an antibody operably connected or otherwise fused to a heterologous Fc portion of a different antibody. In certain embodiments, the Fc domain or heterologous Fc domain is derived from a human. In certain embodiments, the Fc domain or heterologous Fc domain is derived from a mouse. In other embodiments, the heterologous Fc domain may be derived from different Ig classes of the parent antibody, including IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), and IgM. In other embodiments, the heterologous Fc domain may include CH2 domains and CH3 domains from one or more of the different Ig classes. As noted above for humanized antibodies, the antigen-binding fragment of a chimeric antibody may comprise only one or more of the CDRs of an antibody as described herein (e.g., 1, 2, 3, 4, 5, or 6 CDRs of an antibody as described herein), or may comprise entire variable domains (VL, VH, or both).

[0037] Throughout this specification, unless the context requires otherwise, the word "comprise" or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer, or groups of elements or integers, but not the exclusion of any other element or integer, or groups of elements or integers.

[0038] "Consisting of is intended to include and be limited to anything between the phrases "consisting of." Thus, the phrase "consisting of indicates that the listed elements are required or mandatory, and no other elements may be present. "Consisting essentially of is intended to include any element listed between the phrases, and be limited to other elements that do not interfere with or contribute to the activity or function of the listed elements as specified herein. Thus, the phrase "consisting essentially of indicates that the listed elements are required or mandatory, but other elements are optional and may or may not be present depending on whether they substantially affect the activity or function of the listed elements.

[0039] "Immune response" means any immune response originating from the immune system, including responses from cells and humors, innate and adaptive immune systems. Exemplary cellular immune cells include, for example, lymphocytes, macrophages, T cells, B cells, NK cells, neutrophils, eosinophils, dendritic cells, mast cells, monocytes and all subsets thereof. Cellular responses include, for example, effector functions, cytokine release, phagocytosis, efferocytosis, translocation, migration, proliferation, differentiation, activation, inhibition, cell-cell interactions, apoptosis, etc. Humoral responses include, for example, IgG, IgM, IgA, IgE, responses and their corresponding effector functions.

[0040] "Expression control sequences" include nucleic acid or amino acid regulatory sequences, such as promoters, leaders, enhancers, introns, recognition motifs for RNA or DNA binding proteins, polyadenylation signals, terminators, internal ribosome entry sites (IRES), secretion signals, subcellular localization signals, and the like, which can affect the transcription or translation or subcellular or cellular location of a coding sequence in a host cell. Exemplary expression control sequences are described in Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990).

[0041] As used herein, the term "isolated" polypeptide or protein means a protein of the invention that (1) is free of at least some other proteins with which it is normally found in nature, (2) is substantially free of other proteins from the same source (e.g., the same species), (3) is expressed by cells from a different species, (4) has been separated from at least about 50% of the polynucleotides, lipids, carbohydrates, or other substances with which it is associated in nature, (5) is not associated (by covalent or non-covalent interactions) with portions of proteins to which "isolated proteins" are associated in nature, (6) is operatively associated (by covalent or non-covalent interactions) with polypeptides with which it is not associated in nature, or (7) does not occur in nature. Such isolated proteins may be encoded by genomic DNA, cDNA, mRNA or other RNA, may be of synthetic origin, or any combination thereof. In certain embodiments, an isolated protein is substantially free of proteins or polypeptides or other contaminants found in its natural environment that would interfere with its use (therapeutic, diagnostic, prophylactic, research, or otherwise).

[0042] Certain embodiments include biologically active "variants" and "fragments" of the polypeptides (e.g., antibodies) described herein, and polynucleotides encoding the same. "Variants" contain one or more substitutions, additions, deletions, and / or insertions relative to a reference polypeptide or polynucleotide (see, e.g., Tables and Sequence Listings). Variant polypeptides or polynucleotides comprise an amino acid or nucleotide sequence having at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity or similarity or homology to a reference sequence as described herein, and substantially retain an activity of that reference sequence. Also included are sequences that consist of a reference sequence or differ from a reference sequence by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150 or more amino acid or nucleotide additions, deletions, insertions or substitutions and that substantially retain the activity of that reference sequence. In certain embodiments, additions or deletions comprise C-terminal and / or N-terminal additions and / or deletions.

[0043] As used herein, the term "sequence identity" or, for example, comprising "a sequence that is at least 50% identical to..." refers to the degree to which sequences are identical on a nucleotide-by-nucleotide basis or an amino acid-by-amino acid basis within a comparison window. Thus, the "percentage of sequence identity" can be calculated by comparing two optimally aligned sequences within a comparison window, determining the number of positions at which identical nucleic acid bases (e.g., A, T, C, G, I) or identical amino acid residues (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys, and Met) appear in the two sequences to obtain the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window (i.e., the window size), and multiplying the result by 100 to obtain the percentage of sequence identity. Optimal sequence alignment for alignment over a comparison window can be performed by computerized implementation of an algorithm (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Version 7.0, Genetics Computer Group, 575 Science Drive Madison, Wis., USA) or by testing and selecting the best alignment (i.e., that produces the highest homology percentage over the comparison window) produced by any of the various methods selected. Reference can also be made to the BLAST family of programs, as disclosed in, for example, Altschul et al., Nucl. Acids Res. 25:3389, 1997.

[0044] The terms "individual," "subject," and "patient" are used interchangeably herein and refer to any subject in need of treatment or therapy. The subject can be a mammalian subject. Mammalian subjects include, for example, humans, non-human primates, rodents (e.g., rats, mice), lagomorphs (e.g., rabbits), ungulates (e.g., cattle, sheep, pigs, horses, goats, etc.), etc. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human primate, such as a cynomolgus macaque. In some embodiments, the subject is a companion animal (e.g., a cat, a dog).

[0045] As used herein, the terms "therapeutically effective amount," "therapeutic dose," "prophylactically effective amount," or "diagnostically effective amount" are the amount of an agent (e.g., an anti-IL-18BP antibody, immunotherapeutic agent) required to elicit a desired biological response upon administration.

[0046] As used herein, "treatment" of a subject (e.g., a mammal, such as a human primate or non-human primate) or cell is any type of intervention used to attempt to alter the natural course of a disease or condition. Treatment includes, but is not limited to, the administration of a pharmaceutical composition, and can be performed prophylactically or following the initiation of a pathological event or exposure to a pathogen. Also included are "preventative" treatments that may be directed to reducing the rate of progression of the disease or condition being treated, delaying the onset of that disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate complete eradication, cure, or prevention of a disease or condition or its associated symptoms.

[0047] Anti-IL-18BP antibody

[0048] Certain embodiments include antibodies that bind to IL-18BP. In some embodiments, the antibodies modulate (e.g., interfere with, antagonize, inhibit) the binding of IL-18BP to its ligand, interleukin 18 (IL-18). In certain embodiments, the antibodies are characterized by or comprise a heavy chain variable region (V H ), which comprises a complementarity determining region V H CDR1, V H CDR2 and V H CDR3 sequence; and light chain variable region (V L ), which comprises a complementarity determining region V L CDR1, V L CDR2 and V L CDR3 sequence. Illustrative V H , V H CDR1, V H CDR2, V H CDR3; V L, V L CDR1, V L CDR2 and V L The CDR3 sequences are provided in Table A1 and Table A2 below.

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058] Thus, in certain embodiments, the antibody comprises a V H A sequence comprising a complementarity determining region V selected from Table A1 H CDR1, V H CDR2 and V H CDR3 sequences and variants thereof that bind to IL-18BP; and V L A sequence comprising a complementarity determining region V selected from Table A1 L CDR1, V L CDR2 and V L CDR3 sequences and variants thereof that bind to IL-18BP. In certain embodiments, the antibody comprises V H Sequence comprising V H CDR1, V H CDR2 and V H CDR3 sequence; and V L Sequence comprising V L CDR1, V L CDR2 and V L CDR3 sequences, where all CDR sequences are from a single named antibody in Table A1 (e.g., SA04a).

[0059] In certain embodiments, the CDR sequences are as follows:

[0060] The V H CDR1, V H CDR2 and VH The CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 4-6, respectively;

[0061] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 7-9, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 10-12, respectively;

[0062] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 13-15, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 16-18, respectively;

[0063] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 19-21, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 22-24, respectively;

[0064] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 28-30, respectively;

[0065] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 31-33, respectively, and the V L CDR1, V LCDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 34-36, respectively;

[0066] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 37-39, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 40-42, respectively;

[0067] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 46-48, respectively;

[0068] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 49-51, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 52-54, respectively;

[0069] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 58-60, respectively;

[0070] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 64-66, respectively;

[0071] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 70-72, respectively;

[0072] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 73-75, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 76-78, respectively;

[0073] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 79-81, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 82-84, respectively;

[0074] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 85-87, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 88-90, respectively;

[0075] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 91-93, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 94-96, respectively;

[0076] The V H CDR1, V H CDR2 and V HThe CDR3 sequences comprise SEQ ID NOs: 97-99, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 100-102, respectively;

[0077] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 103-105, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 106-108, respectively;

[0078] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 112-114, respectively;

[0079] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 118-120, respectively;

[0080] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 124-126, respectively;

[0081] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and the V LCDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 130-132, respectively;

[0082] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 136-138, respectively;

[0083] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 142-144, respectively;

[0084] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 148-150, respectively;

[0085] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 154-156, respectively;

[0086] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and the V L CDR1, V L CDR2 and V LThe CDR3 sequences comprise SEQ ID NOs: 160-162, respectively;

[0087] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 166-168, respectively;

[0088] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 172-174, respectively;

[0089] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 178-180, respectively;

[0090] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 184-186, respectively;

[0091] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 190-192, respectively;

[0092] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 193-195, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 196-198, respectively;

[0093] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 199-201, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 202-204, respectively;

[0094] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 205-207, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 208-210, respectively;

[0095] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 211-213, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 214-216, respectively;

[0096] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 217-219, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 220-222, respectively;

[0097] The V H CDR1, V HCDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 223-225, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 226-228, respectively;

[0098] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 229-231, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 232-234, respectively;

[0099] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 235-237, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 238-240, respectively;

[0100] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 241-243, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 244-246, respectively;

[0101] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 250-252, respectively;

[0102] The V H CDR1, V H CDR2 and V HThe CDR3 sequences comprise SEQ ID NOs: 253-255, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 256-258, respectively;

[0103] The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 259-261, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 262-264, respectively;

[0104] Variants of the aforementioned CDRs are also included. Exemplary variants bind to IL-18BP and in any one or more of the individual CDRs (e.g., V H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2 and / or V L The present invention relates to a method for the expression of a single CDR3 sequence in a subject. The method comprises the following steps: (1) having 1, 2 or 3 total alterations in any one or more of the CDR3 sequences. Exemplary "alterations" include amino acid substitutions, additions and deletions.

[0105] Illustrative V H and V L Sequence and V H / V L The sequence pairs are provided in Table A2 below. (CDRs are underlined)

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116] Thus, in certain embodiments, the antibody binds to IL-18BP and comprises a V selected from Table A2. H Sequence and corresponding V L In certain embodiments, V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, including, for example, wherein V H There are at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 changes in one or more framework regions. L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, including, for example, wherein V L In certain embodiments, V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, and V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2 and H The regions are from the same single named antibody (e.g., SA04a). In certain embodiments, V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, and V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2 and is from the same single named antibody (e.g., SA04a) as the VH region, wherein no changes are found in the CDRs, as indicated by underlining in Table A2. Thus, the antibody may comprise a VH region. H and V L A sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to the corresponding sequence from a single-named antibody in Table A2 (e.g., SA04a), wherein the antibody comprises the CDRs of the single-named antibody (e.g., SA04a) as listed in Table A1.

[0117] In some embodiments, the VH and V L as follows:

[0118] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 265, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 266;

[0119] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 267, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 268;

[0120] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 269, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 270;

[0121] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 271, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 272;

[0122] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 273, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 274;

[0123] The VH comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 275, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 276;

[0124] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 277, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 278;

[0125] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 279, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 280;

[0126] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 281, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 282;

[0127] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 283, and wherein said V L comprising a sequence at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 284;

[0128] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 285, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 286;

[0129] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 287, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 288;

[0130] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 289, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 290;

[0131] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 291, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 292;

[0132] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 293, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 294;

[0133] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 295, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 296;

[0134] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 297, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 298;

[0135] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 299, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 300;

[0136] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 301, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 302;

[0137] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 303, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 304;

[0138] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 305, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 306;

[0139] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 307, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308;

[0140] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 309, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 310;

[0141] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 311, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 312;

[0142] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 313, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 314;

[0143] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 315, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 316;

[0144] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 317, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 318;

[0145] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 319, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 320;

[0146] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 321, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 322;

[0147] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 323, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 324;

[0148] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 325, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 326;

[0149] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 327, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 328;

[0150] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 329, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 330;

[0151] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 331, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 332;

[0152] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 333, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 334;

[0153] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 335, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 336;

[0154] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 337, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 338;

[0155] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 339, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 340;

[0156] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 341, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 342;

[0157] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 343, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 344;

[0158] The V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 345, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 346;

[0159] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 347, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 348;

[0160] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 349, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 350; or

[0161] The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 351, and wherein said V L Comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO:352.

[0162] It also includes variants thereof that bind to IL-18BP, such as those in the aforementioned V H and / or V L Variants having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in one or more framework regions of any one or more of the sequences. Exemplary "alterations" include amino acid substitutions, additions and deletions.

[0163] As noted above, the antibodies described herein bind to IL-18BP. In certain embodiments, the antibodies bind to human IL-18BP, cynomolgus macaque IL-18BP, and / or mouse IL-18BP, or a region or fragment or epitope thereof.

[0164] Human interleukin-18 binding protein, or IL-18BP, is encoded by the IL18BP gene (see Gene ID: 10068; and UniProt: 095998) and has at least three isoforms. In some embodiments, the antibodies of the present invention bind to isoform A of IL-18BP. In some embodiments, the antibodies of the present invention bind to isoform B of IL-18BP. In some embodiments, the antibodies of the present invention bind to isoforms A and C of IL-18BP. In some embodiments, the antibodies of the present invention bind to all isoforms of IL-18BP. It is an inhibitor of early Th1 cytokine responses and the proinflammatory cytokine IL-18. For example, IL-18BP binds to IL-18, inhibits the binding of IL-18 to its receptor, and thereby inhibits IL-18-induced IFN-γ production, as well as other IL-18 signaling activities. The amino acid sequences of human, cynomolgus macaque, and mouse IL-18BP isoforms are provided in Table B1 below. The signal peptides are underlined in the table below.

[0165]

[0166] Thus, in certain embodiments, the antibody binds to the mature IL-18BP sequence in Table Bl, eg, at a region excluding the signal peptide (underlined).

[0167] In certain embodiments, the antibodies bind to a conformational epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In exemplary embodiments, the antibodies of the invention bind to at least two residues selected from the group consisting of T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In exemplary embodiments, the antibodies of the invention bind to residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372. In exemplary embodiments, such antibodies comprise sequence V H , which comprises at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 347, and sequence V L, which comprises at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 348. In exemplary embodiments, the VHCDR1, VHCDR2 and VHCDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the VLCDR1, VLCDR2 and VLCDR3 sequences comprise SEQ ID NOs: 250-252, respectively.

[0168] In certain embodiments, the antibodies bind to an epitope comprising the IL-18 binding interface of the mature form of IL-18BP. The residues on IL-18BP that interact with IL-18 have been identified as: R61, Y69, S75, H79, T116, S119, and R131. In exemplary embodiments, the antibodies of the invention bind to residues S75, H79, T116, S119, which are also recognized by IL-18. In exemplary embodiments, such antibodies comprise sequence V H , which comprises at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 347, and sequence V L , which comprises at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 348. In exemplary embodiments, the VHCDR1, VHCDR2 and VHCDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the VLCDR1, VLCDR2 and VLCDR3 sequences comprise SEQ ID NOs: 250-252, respectively.

[0169] In certain embodiments, the antibody binds to a linear epitope of the mature IL-18BP sequence of SEQ ID NO: 372 (mature human isoform A). In certain embodiments, the antibody has orthologous specificity or orthologous cross-reactivity for IL-18BP. For example, in certain embodiments, the antibody binds to human IL-18BP and cynomolgus macaque IL-18BP, but does not bind (specifically or substantially) to mouse IL-18BP. In some embodiments, the antibody binds to human IL-18BP, cynomolgus macaque IL-18BP, and mouse IL-18BP.

[0170] In some embodiments, the antibody binds to IL-18 with a binding affinity greater than the binding affinity between IL-18 and IL-18BP (K for human IL-18 and IL-18BP). Dor a specificity measurement of 655 ± 136 pM). In some cases, the antibody binds to human IL-18BP with a stronger binding affinity than about 1 pM to about 10 pM to about 600 pM or 65 pM, or about, at least about, or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM, or optionally with an affinity ranging from about 1 pM to about 600 pM, 1 pM to about 500 pM, 1 pM to about 400 pM, 1 pM to about 300 pM, about 1 pM to about 200 pM, about 1 pM to about 100 pM, about 1 pM to about 50 pM, about 1 pM to about 40 pM, about 1 pM to about 30 pM, about 1 pM to about 20 pM, about 1 pM to about 10 pM, about 1 pM to about 5 pM, about 5 pM to about 600 pM, about 5 pM to about 500 pM, about 5 pM to about 400 pM, about 5 pM to about 300 pM, about 5 pM to about 200 pM, about 5 pM to about 100 pM, about 5 pM to about 50 pM, about 5 pM to about 40 pM, about 5 pM to about 30 pM, about 5 pM to about 20 pM, about 5 pM to about 10 pM, about 10 pM to about 600 pM, about 10 pM to about 500 pM, about 10 pM to about 400 pM, about 10 pM to about 300 pM, about 10 pM to about 200 pM, about 10 pM to about 100 pM, about 10 pM to about 50 pM, about 10 pM to about 40 pM, about 10 pM to about 30 pM, about 10 pM to about 20 pM, or about 20 pM to about 600 pM, about 20 pM to about pM, about 30 pM to about 40 pM, about 30 pM to about 300 pM, about 30 pM to about 200 pM, about 30 pM to about 100 pM, about 20 pM to about 50 pM, about 20 pM to about 40 pM, about 20 pM to about 30 pM, or about 30 pM to about 600 pM, about 30 pM to about 500 pM, about 30 pM to about 400 pM, about 30 pM to about 300 pM, about 30 pM to about 200 pM, about 30 pM to about 100 pM, about 30 pM to about 50 pM, or about 30 pM to about 40 pM. DThis can be determined by biomembrane interferometry (BLI) analysis as described herein. For example, binding kinetic measurements are obtained on a Fortébio (now Sartorius) Octet RED96e instrument by loading the mAb onto an anti-human constant domain (AHC) biosensor (FortéBio) in 10× kinetic buffer consisting of PBS containing 0.1% BSA, 0.02% Tween 20 for 90-120 seconds to achieve a spectral shift value between 0.8 and 1.2 nm. Association can then be performed in the presence of a 2-fold dilution series of hIL-18BP and allowed to proceed for 90-120 seconds. Dissociation can be measured for 300 to 1200 seconds. For weaker variants, the dilution series can start at 100 nM, or for the most potent mAb, at 10 nM.

[0171] In some embodiments, the antibody or its antigen-binding fragment is an IL-18BP antagonist. In some cases, the antibody or its antigen-binding fragment antagonizes the binding and / or signaling activity between IL-18BP and its ligand IL-18. In some embodiments, the antibody or its antigen-binding fragment antagonizes or reduces the binding and / or signaling activity between IL-18BP and IL-18 by about or at least about 10%-1000% (e.g., about 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% or more), for example, in a cell-based assay. In some cases, antagonistic anti-IL-18BP antibodies or antigen-binding fragments thereof block the inhibitory activity of IL-18BP on IL-18, thereby increasing IL-18-mediated signaling, such as IL-18-mediated induction of IFN-γ, CXCL10, and TNFα. These functional activities can be measured by assays disclosed herein. For example, the antibody can be incubated with IL-18BP (e.g., human IL-18BP) followed by addition of IL-18 (e.g., recombinant human IL-18). The resulting solution can then be added to IL-18 reporter HEK 293 cells, which respond to externally added IL-18 by expressing an NF-κB / AP-1 inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. The effect of the antibody can then be analyzed by comparing the effect on the reporter cells to that of a suitable control (e.g., an isotype control antibody). Further details are disclosed in the Materials and Methods section herein. Another possible assay involves measuring the abrogation of the inhibitory effect of anti-IL-18BP mAbs on IFNγ expression by KG-1 cells. Briefly, IL-18BP can be pre-blocked with serial dilutions of the antibody, IL-18 can then be added to the mixture, and this mixture can be added to KG-1 cells and incubated. Secreted IFN-γ can then be measured by conventional means (such as ELISA), and the effect of the test antibody can be compared to the effect of a suitable control (such as an isotype control antibody). Further details are disclosed in the Materials and Methods section herein. Another possible analysis involves incubating PBMCs with the test antibody, IL-12, and IL-18 and measuring IFNγ and / or CCL2 by conventional means. The effect of the test antibody can be compared to the effect of a suitable control (such as an isotype control antibody). Further details are disclosed in the Materials and Methods section herein. Another analysis involves incubating NK cells and pre-complexed hIL-18 / hIL-18BP, followed by the addition of IL-12, and then adding serial dilutions of the test antibody. The effect of the test antibody can be compared to the effect of a suitable control (such as an isotype control antibody). Further details are disclosed in the Materials and Methods section herein.

[0172] Certain embodiments include methods of screening anti-IL-18BP antibodies for the ability to block or inhibit binding between IL-18 and IL-18BP, comprising (a) determining the binding affinity of the antibody to (i) IL-18BP alone and (ii) a low IL-18 fusion protein, wherein the low IL-18 fusion protein comprises IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein binds to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein; (b) comparing the binding affinity of (i) with the binding affinity of (ii); and (c) if the binding affinity of (i) is significantly stronger than the binding affinity of (ii), identifying or selecting the antibody as capable of blocking or inhibiting binding between IL-18 and IL-18BP. Certain embodiments comprise (c) if the binding affinity of (i) is about or at least about 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 200-fold, 300-fold, 400-fold, 500-fold, 600-fold, 700-fold, 800-fold, 900-fold or 1000-fold or more greater than the binding affinity of (ii), then the antibody is identified or selected as capable of blocking or inhibiting the binding between IL-18 and IL-18BP. In certain embodiments, the IL-18 and the IL-18BP are mouse IL-18 and mouse IL-18BP. In some embodiments, the IL-18 and the IL-18BP are human IL-18 and human IL-18BP. In some embodiments, the low IL-18 fusion protein comprises a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and an IL-18BP in an N-terminal to C-terminal orientation. In specific embodiments, the low IL-18 fusion protein comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to a sequence from Table S1.

[0173] For illustrative purposes only, various conventional methods may be used, including Assays (e.g., using appropriately labeled soluble reagents bound to a sensor chip), FACS analysis using cells expressing IL-18BPr on the cell surface (native or recombinant), immunoassays, fluorescent staining assays, ELISA assays, and microcalorimetric methods, such as isothermal titration calorimetry (ITC), can be used to detect and quantify the binding interaction (e.g., binding affinity) between any combination of IL-18BP, IL-18 (e.g., low IL-18), and / or anti-IL-18BP antibodies described herein, or the binding / signaling between IL-18BP and IL-18. Similarly, the functional properties of anti-IL-18BP antibodies can be assessed using a variety of methods known to those skilled in the art: affinity / binding assays (e.g., surface plasmon resonance, competitive inhibition assays); cytotoxicity assays, cell viability assays, cell proliferation or differentiation assays, and inhibition of cancer cell and / or tumor growth using in vitro or in vivo models. Other assays can test the ability of the antibodies described herein to modulate (e.g., inhibit) IL-18BP and / or IL-18-mediated responses. The antibodies described herein can also be tested for in vitro and in vivo efficacy. Such analyses can be performed using recognized protocols known to those skilled in the art (see, for example, Current Protocols in Molecular Biology (Greene Publ. Assoc. Inc. & John Wiley & Sons, Inc., NY, NY); Current Protocols in Immunology (Editors: John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strober 2001 John Wiley & Sons, NY, NY); or commercially available kits.

[0174] In certain embodiments, the Fc region of an antibody comprises, consists of, or consists essentially of an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3, and IgG4), or IgM Fc domain, optionally a human Fc domain, or a hybrid and / or variant thereof. In certain embodiments, the Fc region comprises, consists of, or consists essentially of an Fc from human IgG1 or IgG4 (see, e.g., Allberse and Schuurman, Immunology 105:9-19, 2002), or a fragment or variant thereof.

[0175] In certain embodiments, the antibodies comprise variant or otherwise modified Fc regions, including those with altered properties or biological activities relative to the wild-type Fc region. Examples of modified Fc regions include those with mutant sequences, e.g., by substitution, insertion, deletion, or truncation of one or more amino acids relative to the wild-type sequence; hybrid Fc polypeptides composed of domains from different immunoglobulin classes / subclasses; Fc polypeptides with altered glycosylation / sialylation patterns; and Fc polypeptides modified or derivatized, e.g., by biotinylation (see, e.g., U.S. Application No. 2010 / 0209424), phosphorylation, sulfation, etc., or any combination thereof. Such modifications can be used to alter (e.g., increase, decrease) the binding properties of the Fc region for one or more specific FcRs (e.g., FcγRI, FcγRIIa, FcγRIIb, FcγRIIc, FcγRIIIa, FcγRIIIb, FcRn), its pharmacokinetic properties (e.g., stability or half-life, bioavailability, tissue distribution, volume of distribution, concentration, elimination rate constant, elimination rate, area under the curve (AUC), clearance, C max 、T max 、C min , fluctuation), its immunogenicity, its complement binding or activation and / or CDC / ADCC / ADCP related activities of the Fc region and other properties described herein. Modified Fc regions of human and / or mouse origin are included.

[0176] In certain embodiments, the antibody comprises a hybrid Fc region, for example, an Fc region comprising a combination of Fc domains (e.g., hinge, CH2, CH3, CH4) of immunoglobulins from different species (e.g., human, mouse), different Ig classes, and / or different Ig subclasses. Also included are antibodies comprising a derivatized or otherwise modified Fc region. In certain aspects, the Fc region is modified by, for example, phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, acetylation, amidation, etc. relative to a wild-type or naturally occurring Fc region. In certain embodiments, the Fc region comprises a wild-type or native glycosylation pattern, or alternatively, it comprises increased glycosylation relative to the native form, reduced glycosylation relative to the native form, or it is completely deglycosylated. As an example of a modified Fc glycoform, reduced glycosylation in the Fc region can reduce binding to the C1q region of the first complement component C1, reduce ADCC-related activity, and / or reduce CDC-related activity. Therefore, certain embodiments employ a deglycosylated or non-glycosylated Fc region. For the generation of exemplary aglycosylated Fc regions, see, e.g., WO 2005 / 047337. Another example of an Fc region glycoform is generated according to the numbering system of Kabat et al. by replacing position Q295 with a cysteine ​​residue (see, e.g., U.S. application No. 2010 / 0080794). Certain embodiments include an Fc region in which approximately 80%-100% of the glycoproteins in the Fc region comprise a mature core carbohydrate structure lacking fucose (see, e.g., U.S. application No. 2010 / 0255013). Some embodiments include an Fc region that is optimized by substitution or deletion to reduce the level of fucosylation, e.g., to increase affinity for Fc RI, Fc RIa, or Fc RIIIa, and / or to improve phagocytosis by cells expressing Fc RIIa (see, e.g., U.S. application Nos. 2010 / 0249382 and 2007 / 0148170).

[0177] As another example of a modified Fc glycoform, the Fc region of an antibody may comprise oligomannose-type N-glycans and, optionally, have one or more of the following: increased ADCC effector activity, increased binding affinity for FcγRIIIA (and certain other FcRs), similar or increased binding specificity for a target of an IL-18BP polypeptide, similar or higher binding affinity for a target of an IL-18BP polypeptide, and / or similar or lower binding affinity for the mannose receptor relative to a corresponding Fc region containing complex-type N-glycans (see, e.g., U.S. Application No. 2007 / 0092521 and U.S. Patent No. 7,700,321). As another example, enhanced affinity of the Fc region for FcγRs has been achieved using engineered glycoforms produced by expressing antibodies in engineered or mutant cell lines (see, e.g., Umana et al., Nat Biotechnol. 17:176-180, 1999; Davies et al., Biotechnol Bioeng. 74:288-294, 2001; Shields et al., J Biol Chem. 277:26733-26740, 2002; Shinkawa et al., 2003, J Biol Chem. 278:3466-3473, 2003; and U.S. Application No. 2007 / 0111281). Certain Fc region glycoforms comprise an increased proportion of N-glycosidically bonded complex carbohydrate chains, with fucose at the reducing end of the carbohydrate chain not bound to the N-acetylglucosamine at the 6-position (see, e.g., U.S. Application No. 2010 / 0092997). Specific embodiments may include IgG Fc regions glycosylated with at least one galactose moiety linked to a respective terminal sialic acid moiety via an α-2,6 linkage, optionally wherein the Fc region has increased anti-inflammatory activity relative to a corresponding wild-type Fc region (see, e.g., U.S. Application No. 2008 / 0206246). Certain of these and related altered glycosylation approaches have significantly enhanced the ability of an Fc region to selectively bind to an FcR, such as FcγRIII, mediate ADCC, and alter other properties of the Fc region as described herein.

[0178] Relative to the corresponding wild-type Fc sequence (e.g., same species, same Ig class, same Ig subclass), certain variants, fragments, hybrids or otherwise modified Fc regions of antibodies may have altered binding to one or more FcRs, and / or corresponding changes in effector function. For example, relative to the corresponding wild-type Fc sequence, such Fc regions may increase binding to one or more of Fcγ receptors, Fcα receptors, Fcε receptors and / or neonatal Fc receptors. In other embodiments, relative to the corresponding wild-type Fc sequence, the binding of variants, fragments, hybrids or modified Fc regions to one or more of Fcγ receptors, Fcα receptors, Fcε receptors and / or neonatal Fc receptors may be reduced. Specific FcRs are described elsewhere herein.

[0179] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations relative to the corresponding wild-type Fc sequence to increase binding to one or more of an Fcγ receptor, an Fcα receptor, an Fcε receptor, and / or a neonatal Fc receptor. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations relative to the corresponding wild-type Fc sequence to increase binding to one or more of an Fcγ receptor, an Fcα receptor, an Fcε receptor, and / or a neonatal Fc receptor. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to increase effector function. In some embodiments, at least one antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function.

[0180] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function. In some embodiments, the antibody is a partially blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the partially blocking antibody comprises an Fc domain selected from human IgG1 and IgG3 comprising one or more mutations to increase effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with high effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG1 or IgG3 comprising one or more mutations to increase effector function.

[0181] In some embodiments, the antibody comprises an Fc domain comprising one or more mutations relative to the corresponding wild-type Fc sequence to reduce binding to one or more of an Fcγ receptor, an Fcα receptor, an Fcε receptor, and / or a neonatal Fc receptor. In some embodiments, the antibody comprises an IgG1 or IgG3 Fc domain comprising one or more mutations relative to the corresponding wild-type Fc sequence to reduce binding to one or more of an Fcγ receptor, an Fcα receptor, an Fcε receptor, and / or a neonatal Fc receptor. In some embodiments, the antibody comprises an Fc domain comprising one or more mutations to reduce effector function. In some embodiments, the antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function.

[0182] In some embodiments, the antibody is a blocking antibody comprising an Fc domain with low effector activity. In some embodiments, the blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a partially blocking antibody comprising an Fc domain with low effector activity. In some embodiments, the partially blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function. In some embodiments, the antibody is a non-blocking antibody comprising an Fc domain with low effector activity. In some embodiments, the non-blocking antibody comprises an Fc domain selected from human IgG2 and IgG4 comprising one or more mutations to reduce effector function.

[0183] Specific examples of Fc variants with altered (e.g., increased, decreased) effector function / FcR binding can be found in, for example, U.S. Patent Nos. 5,624,821 and 7,425,619; U.S. Application Nos. 2009 / 0017023, 2009 / 0010921, and 2010 / 0203046; and WO 2000 / 42072 and WO 2004 / 016750. Certain examples include human Fc regions with one or more substitutions at positions 298, 333, and / or 334, such as S298A, E333A, and / or K334A (numbering based on the EU index of Kabat et al.), which have been shown to increase binding to the activating receptor FcγRIIIa and decrease binding to the inhibitory receptor FcγRIIb. These mutations can be combined to obtain double and triple mutant variants with further improved FcR binding. Certain embodiments include the S298A / E333A / K334A triple mutant, which increases binding to FcγRIIIa, decreases binding to FcγRIIb, and increases ADCC (see, e.g., Shields et al., J Biol Chem. 276:6591-6604, 2001; and Presta et al., Biochem Soc Trans. 30:487-490, 2002). See also, for example, Umana et al., supra; and U.S. Patent No. 7,662,925, for the engineered Fc glycoforms with increased binding to FcRs. Some embodiments include Fc regions comprising one or more substitutions selected from 434S, 252Y / 428L, 252Y / 434S, and 428L / 434S based on the EU index as in Kabat et al. (see U.S. application Ser. Nos. 2009 / 0163699 and 20060173170). Some embodiments include Fc regions comprising one or more substitutions selected from L234A, L235A, and G237A based on the EU index as in Kabat et al. (see U.S. application Ser. No. 17 / 779,425). Some embodiments include Fc regions comprising substitutions at L234A and L235A based on the EU index as in Kabat et al. Some embodiments include Fc regions comprising substitutions at L234A and G237A based on the EU index as in Kabat et al. Some embodiments include Fc regions comprising substitutions at L235A and G237A, based on the EU index as in Kabat et al. Some embodiments include Fc regions comprising substitutions at L234A, L235A, and G237A, based on the EU index as in Kabat et al. Some embodiments include Fc regions comprising one or more substitutions selected from the group consisting of M252Y, S254T, and T256E, based on the EU index as in Kabat et al.Some embodiments include Fc regions comprising one or more substitutions selected from M428L and N434S based on the EU index of Kabat et al. In some embodiments, the Fc substitutions disclosed herein are for an Fc domain selected from human IgG1, IgG2, IgG3, or IgG4. In some embodiments, the Fc substitutions disclosed herein are for a human IgG1 Fc domain. In some embodiments, the Fc substitutions disclosed herein are for a human IgG2 Fc domain. In some embodiments, the Fc substitutions disclosed herein are for a human IgG3 Fc domain. In some embodiments, the Fc substitutions disclosed herein are for a human IgG4 Fc domain. In some embodiments, the antibodies of the invention comprise one or more Fc substitutions disclosed herein and V. H and V L A sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to the corresponding sequence from a single named antibody in Table A2 (e.g., SA04a), wherein the antibody comprises the CDRs of the single named antibody (e.g., SA04a) as listed in Table A1.

[0184] Relative to the corresponding wild-type Fc sequence, certain variant hybrids or modified Fc regions may have altered solubility. In certain embodiments, such Fc regions may have increased solubility relative to the corresponding wild-type Fc sequence. In other embodiments, variant hybrids or modified Fc regions may have reduced solubility relative to the corresponding wild-type Fc sequence. Solubility can be measured, for example, in vitro (e.g., under physiological conditions) according to conventional techniques in the art. Exemplary solubility measurements are described elsewhere herein.

[0185] Variant Fc regions may also have one or more mutant hinge regions as described, for example, in U.S. Application No. 2003 / 0118592. For example, one or more cysteines in the hinge region may be deleted or substituted with different amino acids. The mutant hinge region may contain no cysteine ​​residues, or it may contain 1, 2, or 3 fewer cysteine ​​residues than the corresponding wild-type hinge region. In some embodiments, Fc regions having such mutant hinge regions exhibit reduced dimerization ability relative to wild-type Ig hinge regions.

[0186] In some embodiments, the antibodies or antigen-binding fragments thereof may be combined with one or more cytotoxic or chemotherapeutic agents. In some embodiments, the antibodies disclosed herein are combined with or operably linked to a radioactive isotope to form a radioconjugate and / or macrocyclic chelator suitable for binding radioactive metal ions. The antibodies can be used in any of the compositions, methods, and / or kits described herein and in combination with one or more of the additional agents described herein.

[0187] Methods of use and pharmaceutical compositions

[0188] Certain embodiments relate to methods for treating a disease or condition in a subject in need thereof, improving the symptoms of the disease or condition and / or reducing the progression of the disease or condition, comprising administering to the subject an antibody bound to IL-18BP as described herein or a pharmaceutical composition comprising the same. Also included is a method for stimulating an immune response (e.g., an IL-18 mediated immune response) in a subject in need thereof, comprising administering to the subject a pharmaceutical composition as described herein. In some cases, the antibody antagonizes the binding / signaling activity between IL-18BP and its ligand IL-18, and thereby increases IL-18 mediated signaling or activity (e.g., increased induction of IFN-γ, CXCL10, and / or TNFα). In some embodiments, as noted above, the disease or condition is a cancer or tumor, or an infectious disease. In some embodiments, the disease is any disease in which immune system activation may be beneficial.

[0189] In some embodiments, as noted above, the disease or condition is a cancer or tumor or other proliferative disease or disorder, such as a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enteritis, a proliferative diabetic retinopathy, or a proliferative nephropathy. In some cases, the cancer or tumor expresses or overexpresses IL-18BP, IL-18, or both. In some cases, the proliferative disease or disorder is associated with increased expression of IL-18BP, IL-18, or both. In some cases, the cancer is a primary cancer. In some cases, the cancer is a metastatic cancer. Certain embodiments therefore include methods of treating cancer in a patient in need thereof, reducing the severity of cancer, or preventing cancer, comprising administering to the patient a composition as described herein, including wherein the antibody is an IL-18BP antagonist, thereby treating the cancer, reducing the severity of the cancer, or preventing the cancer.

[0190] Exemplary cancers include, but are not limited to, bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma, Hodgkin's lymphoma), liver cancer (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and gastric cancer. In specific embodiments, the cancer is a metastatic cancer, eg, one that has metastasized to the bones.

[0191] Also provided is an antibody, an antigen-binding fragment thereof, or a pharmaceutical composition of the present invention for use as a medicament. The antibody, fragment thereof, or pharmaceutical composition of the present invention can be used in any of the methods of treatment disclosed herein. In specific embodiments, the antibody, fragment thereof, or pharmaceutical composition of the present invention can be used to treat any disease or condition disclosed herein (such as cancer or tumor or other proliferative disease or condition, such as lymphoproliferative disorder, myeloproliferative disorder, proliferative enteritis, proliferative diabetic retinopathy, or proliferative nephropathy), improve its symptoms, and / or reduce its progression.

[0192] Certain embodiments include combination therapies, e.g., comprising administering a pharmaceutical composition described herein (comprising an anti-IL-18BP antibody) in combination with one or more additional therapeutic agents (e.g., an immunostimulant, an immune checkpoint modulator, and / or a chemotherapeutic agent). In some embodiments, the additional therapeutic agent comprises IL-18, including human IL-18 (or a functional variant or fragment thereof).

[0193] In certain embodiments, additional therapeutic agents include immune checkpoint regulators. Specific examples of immune checkpoint regulators include " antagonists " or " inhibitors " of one or more inhibitory immune checkpoint molecules, and " agonists " of one or more stimulating immune checkpoint molecules. In general, immune checkpoint molecules are components that enhance signals (co-stimulatory molecules) or reduce signals of the immune system, and their targeting has cancer treatment potential because cancer cells can interfere with the natural functions of immune checkpoint molecules (see, for example, Sharma and Allison, Science (Science) 348: 56-61, 2015; Topalian et al., Cancer Cell (Cancer Cell) 27: 450-461, 2015; Pardoll, Nature Reviews (Nature Reviews Cancer) 12: 252-264, 2012). In certain embodiments, immune checkpoint regulators (e.g., antagonists, agonists) are "bound" or "specifically bound" to one or more immune checkpoint molecules, as described herein.

[0194] In certain embodiments, immune checkpoint regulators are polypeptides or peptides. The terms "peptide" and "polypeptide" are used interchangeably herein, however, in some cases, the term "peptide" may refer to shorter polypeptides, for example, polypeptides consisting of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45 or 50 amino acids (including all integers and ranges (e.g., 5-10, 8-12, 10-15)). Polypeptides and peptides can be made up of naturally occurring amino acids and / or non-naturally occurring amino acids as described herein.

[0195] Antibodies are also included as polypeptides. Thus, in some embodiments, the immune checkpoint modulating polypeptide agent is an "antibody" as described herein.

[0196] In some embodiments, the agent is or comprises a "ligand" of an immune checkpoint molecule, such as a natural ligand. "Ligand" generally refers to a substance or molecule that forms a complex with a target molecule (e.g., a biomolecule) for biological purposes, and includes a "protein ligand" that generally generates a signal by binding to a site on a target molecule or target protein. Therefore, some agents are protein ligands that bind to immune checkpoint molecules and generate signals in nature. Also included are "modified ligands", such as protein ligands fused to pharmacokinetic modulators, such as those derived from the Fc region of an immunoglobulin.

[0197] In some embodiments, the pharmaceutical agent is a "small molecule," which refers to an organic compound of synthetic or biological origin (biomolecule), but not typically a polymer. An organic compound refers to a large class of compounds whose molecules contain carbon, typically excluding compounds containing only carbonates, simple carbon oxides, or cyanides. A "biomolecule" generally refers to an organic molecule produced by a living organism, and also includes large polymer molecules (biopolymers), such as peptides, polysaccharides, and nucleic acids, and small molecules, such as major secondary metabolites, lipids, phospholipids, glycolipids, sterols, glycerolipids, vitamins, and hormones. A "polymer" generally refers to a macromolecule or macromolecule composed of repeating structural units, typically linked by covalent chemical bonds.

[0198] In some embodiments, the immune checkpoint modulator is an antagonist or inhibitor of one or more inhibitory immune checkpoint molecules. Exemplary inhibitory immune checkpoint molecules include programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), programmed death 1 (PD-1), cytotoxic T lymphocyte-associated protein 4 (CTLA-4), indoleamine 2,3-dioxygenase (IDO), tryptophan 2,3-dioxygenase (TDO), T cell immunoglobulin domain and mucin domain 3 (TIM-3), lymphocyte activation gene-3 (LAG-3), V-domain Ig inhibitory factor of T cell activation (VISTA), B and T lymphocyte attenuator (BTLA), CD160, and T cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0199] In certain embodiments, the agent is a PD-1 (receptor) antagonist or inhibitor, which has been shown to target and restore immune function in the tumor environment (see, for example, Phillips et al., Int Immunol. 27: 39-46, 2015). PD-1 is a cell surface receptor belonging to the immunoglobulin superfamily and expressed on T cells and progenitor B cells. PD-1 interacts with two ligands, PD-L1 and PD-L2. PD-1 acts as an inhibitory immune checkpoint molecule, for example, by reducing or preventing the activation of T cells, thereby reducing autoimmunity and promoting self-tolerance. The inhibitory effect of PD-1 is achieved at least in part by promoting apoptosis of antigen-specific T cells in lymph nodes and simultaneously reducing apoptosis of regulatory T cells (suppressor T cells). Some examples of PD-1 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-1 and reduce one or more immunosuppressive activities thereof, such as its downstream signaling or its interaction with PD-L1. Specific examples of PD-1 antagonists or inhibitors include the antibodies nivolumab, pembrolizumab, PDR001, MK-3475, AMP-224, AMP-514, and pilizumab, and antigen-binding fragments thereof (see, e.g., U.S. Pat. Nos. 8,008,449, 8,993,731, 9,073,994, 9,084,776, 9,102,727, 9,102,728, 9,181,342, 9,217,034, 9,387,247, 9,492,539, 9,492,540; and U.S. App. Nos. 2012 / 0039906, 2015 / 0203579).

[0200] In some embodiments, the agent is a PD-L1 antagonist or inhibitor. As noted above, PD-L1 is one of the natural ligands of the PD-1 receptor. General examples of PD-L1 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-L1 and reduce one or more immunosuppressive activities thereof, such as its binding to the PD-1 receptor. Specific examples of PD-L1 antagonists include antibodies atezolizumab (MPDL3280A), avelumab (MSB0010718C) and durvalumab (MEDI4736) and antigen-binding fragments thereof (see, for example, U.S. Patent Nos. 9,102,725, 9,393,301, 9,402,899, 9,439,962).

[0201] In some embodiments, the agent is a PD-L2 antagonist or inhibitor. As noted above, PD-L2 is one of the natural ligands of the PD-1 receptor. General examples of PD-L2 antagonists or inhibitors include antibodies or small molecules that specifically bind to PD-L2 and reduce one or more of its immunosuppressive activities, such as its binding to the PD-1 receptor.

[0202] In some embodiments, the agent is a CTLA-4 antagonist or inhibitor. Cytotoxic T lymphocyte-associated protein 4 (CTLA4 or CTLA-4), also known as cluster of differentiation 152 (CD152), is a protein receptor that acts as an inhibitory immune checkpoint molecule by transmitting an inhibitory signal to T cells when it is bound to CD80 or CD86 on the surface of antigen-presenting cells. General examples of CTLA-4 antagonists or inhibitors include antibodies or small molecules that specifically bind to CTLA-4. Specific examples include antibodies ipilimumab and tremelimumab and antigen-binding fragments thereof. It is believed that at least some of the activity of ipilimumab is mediated by killing CTLA-4-expressing inhibitory Tregs through antibody-dependent cell-mediated cytotoxicity (ADCC).

[0203] In some embodiments, the agent is an IDO antagonist or inhibitor, or a TDO antagonist or inhibitor. IDO and TDO are tryptophan catabolism enzymes with immunosuppressive properties. For example, IDO is known to inhibit T cells and NK cells, generate and activate Tregs and myeloid-derived suppressor cells, and promote tumor angiogenesis. General examples of IDO and TDO antagonists or inhibitors include antibodies or small molecules that specifically bind to IDO or TDO (see, e.g., Platten et al., Front Immunol. 5:673, 2014) and reduce or inhibit one or more immunosuppressive activities. Specific examples of IDO antagonists or inhibitors include indomod (NLG-8189), 1-methyl-tryptophan (1MT), β-carboline (norharman; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epasta (see, e.g., Sheridan, Nature Biotechnology. 33:321-322, 2015). Specific examples of TDO antagonists or inhibitors include 680C91 and LM10 (see, eg, Pilotte et al., PNAS USA. 109:2497-2502, 2012).

[0204] In some embodiments, the agent is a TIM-3 antagonist or inhibitor. T cell immunoglobulin domain and mucin domain 3 (TIM-3) is expressed on activated human CD4+ T cells and regulates Th1 and Th17 cytokines. TIM-3 also acts as a negative regulator of Th1 / Tc1 function by triggering cell death when interacting with its ligand galectin-9. TIM-3 contributes to the suppression of the tumor microenvironment, and its overexpression is associated with poor prognosis in a variety of cancers (see, for example, Li et al., Acta Oncol. 54: 1706-13, 2015). General examples of TIM-3 antagonists or inhibitors include antibodies or small molecules that specifically bind to TIM-3 and reduce or inhibit one or more of its immunosuppressive activities.

[0205] In some embodiments, the agent is a LAG-3 antagonist or inhibitor. Lymphocyte activation gene-3 (LAG-3) is expressed on activated T cells, natural killer cells, B cells and plasmacytoid dendritic cells. It negatively regulates cell proliferation, activation and homeostasis of T cells in a manner similar to CTLA-4 and PD-1 (see, for example, Workman and Vignali., European Journal of Immunology (European Journal of Immun.) 33:970-9, 2003; and Workman et al., Journal of Immunology (Journal of Immunology) 172:5450-5, 2004), and has been reported to play a role in Treg suppressive function (see, for example, Huang et al., Immunity (Immunity) 21:503-13, 2004). LAG3 also maintains CD8+ T cells in a tolerant state and is combined with PD-1 to maintain CD8 T cell exhaustion. General examples of LAG-3 antagonists or inhibitors include antibodies or small molecules that specifically bind to LAG-3 and inhibit one or more of its immunosuppressive activities. Specific examples include the antibody BMS-986016 and antigen-binding fragments thereof.

[0206] In certain embodiments, medicament is VISTA antagonist or inhibitor.The V domain Ig inhibitory factor (VISTA) of T cell activation is mainly expressed on hematopoietic cells, and is to suppress T cell activation, induce Foxp3 expression and be highly expressed in the inhibitory immune checkpoint regulatory factor (see, for example, Lines et al., Cancer Research (Cancer Res.) 74:1924-32,2014) in the tumor microenvironment that suppresses anti-tumor T cell response.General examples of VISTA antagonists or inhibitors include antibodies or small molecules that specifically bind to VISTA and reduce its one or more immunosuppressive activities.

[0207] In some embodiments, the agent is a BTLA antagonist or inhibitor. During T cell activation, the expression of B and T lymphocyte attenuator (BTLA; CD272) is induced, and it inhibits T cells via interaction with tumor necrosis family receptors (TNF-R) and B7 family cell surface receptors. BTLA is a ligand of tumor necrosis factor (receptor) superfamily member 14 (TNFRSF14), also known as herpes virus invasion mediator (HVEM). The BTLA-HVEM complex negatively regulates T cell immune responses, for example, by inhibiting the function of human CD8+ cancer-specific T cells (see, for example, Derré et al., Journal of Clinical Research (J Clin Invest) 120: 157-67, 2009). General examples of BTLA antagonists or inhibitors include antibodies or small molecules that specifically bind to BTLA-4 and reduce one or more of its immunosuppressive activities.

[0208] In some embodiments, the agent is an HVEM antagonist or inhibitor, e.g., an antagonist or inhibitor that specifically binds to HVEM and interferes with its interaction with BTLA or CD 160. General examples of HVEM antagonists or inhibitors include antibodies or small molecules that specifically bind to HVEM, optionally reduce the HVEM / BTLA and / or HVEM / CD160 interaction, and thereby reduce one or more of the immunosuppressive activities of HVEM.

[0209] In some embodiments, the agent is a CD160 antagonist or inhibitor, e.g., an antagonist or inhibitor that specifically binds to CD160 and interferes with its interaction with HVEM. General examples of CD160 antagonists or inhibitors include antibodies or small molecules that specifically bind to CD160, optionally reduce the CD160 / HVEM interaction, and thereby reduce or inhibit one or more of its immunosuppressive activities.

[0210] In some embodiments, the agent is a TIGIT antagonist or inhibitor. T cell Ig and ITIM domain (TIGIT) is a co-inhibitory receptor found on the surface of a variety of lymphocytes and, for example, inhibits anti-tumor immunity via Treg (Kurtulus et al., Journal of Clinical Research (J Clin Invest.) 125: 4053-4062, 2015). General examples of TIGIT antagonists or inhibitors include antibodies or small molecules that specifically bind to TIGIT and reduce one or more of its immunosuppressive activities (see, for example, Johnston et al., Cancer Cell 26: 923-37, 2014).

[0211] In certain embodiments, the immune checkpoint modulator is an agonist of one or more stimulatory immune checkpoint molecules. Exemplary stimulatory immune checkpoint molecules include OX40, CD40, glucocorticoid-induced TNFR family-related genes (GITR), CD137 (4-1BB), CD27, CD28, CD226, and herpes virus entry mediator (HVEM).

[0212] In some embodiments, the agent is an OX40 agonist. OX40 (CD134) promotes the expansion of effector T cells and memory T cells and inhibits the differentiation and activity of T regulatory cells (see, e.g., Croft et al., Immunol Rev. 229: 173-91, 2009). Its ligand is OX40L (CD252). Because OX40 signaling affects both T cell activation and survival, it plays a key role in initiating anti-tumor immune responses in lymph nodes and maintaining anti-tumor immune responses in the tumor microenvironment. General examples of OX40 agonists include antibodies or small molecules or ligands that specifically bind to OX40 and increase its one or more immunostimulatory activities. Specific examples include OX86, OX-40L, Fc-OX40L, GSK3174998, MEDI0562 (humanized OX40 agonist), MEDI6469 (murine OX4 agonist), and MEDI6383 (OX40 agonist), and antigen-binding fragments thereof.

[0213] In some embodiments, the agent is a CD40 agonist. CD40 is expressed on antigen presenting cells (APCs) and some malignant tumors. Its ligand is CD40L (CD154). On APCs, engagement causes upregulation of co-stimulatory molecules, which may not require T cell assistance in anti-tumor immune responses. CD40 agonist therapy plays an important role in APC maturation and its migration from tumors to lymph nodes, causing increased antigen presentation and T cell activation. Anti-CD40 agonist antibodies produce significant responses and lasting anti-cancer immunity in animal models, which is at least in part mediated by cytotoxic T cells (see, for example, Johnson et al., Clin Cancer Res. 21: 1321-1328, 2015; and Vonderheide and Glennie, Clin Cancer Res. 19: 1035-43, 2013). General examples of CD40 agonists include antibodies, small molecules, or ligands that specifically bind to CD40 and enhance one or more of its immunostimulatory activities. Specific examples include sotigalilmab, CP-870,893, dacillinam, Chi Lob 7 / 4, ADC-1013, CD40L, rhCD40L, and antigen-binding fragments thereof.

[0214] In some embodiments, the agent is a GITR agonist. Glucocorticoid-induced TNFR family-related genes (GITR) increase T cell expansion, inhibit the suppressive activity of Treg, and prolong the survival of T effector cells. GITR agonists have been shown to promote anti-tumor responses via the loss of Treg lineage stability (see, for example, Schaer et al., Cancer Immunology Research (Cancer Immunol Res.) 1: 320-31, 2013). These different mechanisms illustrate that GITR plays an important role in initiating immune responses in lymph nodes and maintaining immune responses in tumor tissues. Its ligand is GITRL. General examples of GITR agonists include antibodies or small molecules or ligands that specifically bind to GITR and increase its one or more immunostimulatory activities. Specific examples include GITRL, INCAGN01876, DTA-1, MEDI1873 and its antigen-binding fragment.

[0215] In some embodiments, the agent is a CD137 agonist. CD137 (4-1BB) is a member of the tumor necrosis factor (TNF) receptor family, and the cross-linking of CD137 enhances T cell proliferation, IL-2 secretion, survival and cytolytic activity. CD137-mediated signal transduction also protects T cells such as CD8+ T cells from activation-induced cell death. General examples of CD137 agonists include antibodies or small molecules or ligands that specifically bind to CD137 and increase one or more immunostimulatory activities thereof. Specific examples include CD137 (or 4-1BB) ligands (see, for example, Shao and Schwarz, Journal of Leukocyte Biology (J LeukocBiol.) 89: 21-9, 2011) and antibody utomilumab (utomilumab), including its antigen-binding fragment.

[0216] In some embodiments, the agent is a CD27 agonist. CD27 stimulation increases the antigen-specific expansion of naive T cells and contributes to the long-term maintenance of T cell memory and T cell immunity. Its ligand is CD70. Agonist antibodies target human CD27 to stimulate T cell activation and anti-tumor immunity (see, for example, Thomas et al., Oncoimmunology 2014; 3: e27255.doi: 10.4161 / onci.27255; and He et al., J Immunol. 191: 4174-83, 2013). General examples of CD27 agonists include antibodies or small molecules or ligands that specifically bind to CD27 and increase one or more immunostimulatory activities thereof. Specific examples include CD70 and the antibodies valirumab and CDX-1127 (1F5), including antigen-binding fragments thereof.

[0217] In some embodiments, the agent is a CD28 agonist. CD28 is constitutively expressed by CD4+ T cells and some CD8+ T cells. Its ligands include CD80 and CD86, and its stimulation increases T cell expansion. General examples of CD28 agonists include antibodies, small molecules, or ligands that specifically bind to CD28 and enhance one or more of its immunostimulatory activities. Specific examples include CD80, CD86, the antibody TAB08, and antigen-binding fragments thereof.

[0218] In some embodiments, the agent is a CD226 agonist. CD226 is a stimulatory receptor that shares a ligand with TIGIT, and in contrast to TIGIT, the engagement of CD226 enhances T cell activation (see, for example, Kurtulus et al., Journal of Clinical Research (J Clin Invest.) 125: 4053-4062, 2015; Bottino et al., Journal of Experimental Medicine (J Exp Med.) 1984: 557-567, 2003; and Tahara-Hanaoka et al., International Immunology (Int Immunol.) 16: 533-538, 2004). General examples of CD226 agonists include antibodies or small molecules or ligands (e.g., CD112, CD155) that specifically bind to CD226 and increase its one or more immunostimulatory activities.

[0219] In some embodiments, the agent is an HVEM agonist. Herpes simplex virus invasion mediator (HVEM), also known as tumor necrosis factor receptor superfamily member 14 (TNFRSF14), is a human cell surface receptor of the TNF receptor superfamily. HVEM is found on a variety of cells including T cells, APCs, and other immune cells. Unlike other receptors, HVEM is expressed at high levels on resting T cells and is downregulated upon activation. HVEM signaling has been shown to play a key role in the early stages of T cell activation and during the expansion of tumor-specific lymphocyte populations in lymph nodes. General examples of HVEM agonists include antibodies or small molecules or ligands that specifically bind to HVEM and enhance one or more of its immunostimulatory activities.

[0220] In some embodiments, the additional therapeutic agent comprises a chemotherapeutic agent, such as a small molecule chemotherapeutic agent. Non-limiting examples of chemotherapeutic agents include alkylating agents, antimetabolites, cytotoxic antibiotics, topoisomerase inhibitors (type 1 or type II), anti-microtubule agents, and the like.

[0221] In certain embodiments, the methods and compositions described herein are sufficient to cause tumor regression, as indicated by a statistically significant decrease in the amount of viable tumor, such as a decrease in tumor mass of at least 10%, 20%, 30%, 40%, 50% or greater, or a change in scan size (e.g., a decrease with statistical significance). In some embodiments, the methods and compositions described herein reduce the growth rate of a cancer (e.g., in vivo or in vitro, including cancer cells isolated from a biopsy or other sample and grown in vitro) by about or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or greater relative to an untreated control. In some cases, the methods and compositions described herein reduce cancer cell initiation, migration, adhesion, invasiveness, and / or metastasis by about or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more relative to untreated controls. In some cases, the methods and compositions described herein reduce angiogenesis in a tumor environment by about or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000%, 2000% or more relative to an untreated control.

[0222] In certain embodiments, the disease or condition is myelodysplastic syndrome (MDS) (see, e.g., Wang et al., Blood 140(Suppl 1):12297, 2022), for example, for which antagonizing IL-18BP represents a viable approach. MDS refers to a group of cancers in which immature blood cells in the bone marrow do not mature and, therefore, do not develop into healthy blood cells. Certain embodiments therefore include methods of treating, reducing the severity of, or preventing MDS in a patient in need thereof, comprising administering to the patient a composition described herein, including wherein the antibody is an IL-18BP antagonist, thereby treating, reducing the severity of, or preventing MDS.

[0223] In some embodiments, the disease or condition is an infectious disease. For example, in certain embodiments, the infectious disease is selected from viruses (see, e.g., Vecchie et al., J Cell Physiol. 236(3):1638-1657, 2021), bacteria (see, e.g., Kinoshita et al., Ann Surg. 240(2):313-20, 2004), fungi (e.g., yeast), and protozoan infections. Some embodiments therefore include methods of treating an infectious disease in a patient in need thereof, reducing the severity of an infectious disease, or preventing an infectious disease, comprising administering to the patient a composition as described herein, including wherein the antibody is an IL-18BP antagonist, thereby treating the infectious disease, reducing the severity of the infectious disease, or preventing the infectious disease.

[0224] In certain embodiments, the methods and compositions described herein are sufficient to cause disease stabilization.In certain embodiments, the methods and compositions described herein are sufficient to cause a clinically relevant reduction in symptoms for a specific disease indication known to a skilled clinician.

[0225] For in vivo use, certain embodiments include pharmaceutical compositions comprising an antibody as described herein and a pharmaceutically acceptable carrier. To prepare a therapeutic or pharmaceutical composition, an effective amount or a desired amount of one or more agents is mixed with any pharmaceutical carrier or excipient known to those skilled in the art to be suitable for a particular agent and / or mode of administration. The pharmaceutical carrier can be liquid, semi-liquid, or solid. Solutions or suspensions for parenteral, intradermal, intraocular, subcutaneous, direct instillation into the bladder, or topical administration may include, for example, a sterile diluent (such as water), a physiological saline solution (e.g., phosphate-buffered saline; PBS), a fixed oil, polyethylene glycol, glycerol, propylene glycol, or other synthetic solvents; antibacterial agents (such as benzyl alcohol and methyl paraben); antioxidants (such as ascorbic acid and sodium bisulfite) and chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); buffers (such as acetates, citrates, and phosphates). If administered intravenously (eg, by IV infusion), suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0226] Administration of the agents described herein in pure form or in the form of appropriate therapeutic or pharmaceutical compositions can be carried out via any of the generally accepted modes of administration for similarly acting agents. Therapeutic or pharmaceutical compositions can be prepared by combining a composition containing the agent with an appropriate physiologically acceptable carrier, diluent, or excipient and can be formulated into solid, semisolid, liquid, or gaseous formulations such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. In addition, other pharmaceutically active ingredients (including other small molecules as described elsewhere herein) and / or suitable excipients, such as salts, buffers, and stabilizers, may, but need not, be present in the composition.

[0227] Administration can be achieved by a variety of different routes, including oral, parenteral, nasal, intravenous, intraocular, intradermal, intramuscular, subcutaneous, instillation into the bladder, transdermal, inhalation, sublingual, buccal, rectal, vaginal or topical. The preferred mode of administration depends on the nature of the condition to be treated or prevented. Specific embodiments include administration by IV infusion.

[0228] The carrier may include, for example, a pharmaceutically or physiologically acceptable carrier, excipient, or stabilizer that is nontoxic to the cells or mammals exposed thereto at the dosages and concentrations employed. Typically, a physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include: buffers such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, histidine, and / or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as polysorbate 20 (TWEEN TM ), polyethylene glycol (PEG) and poloxamer (PLURONICS TM )wait.

[0229] In some embodiments, one or more pharmaceutical agents may be encapsulated in microcapsules (e.g., hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively) prepared, for example, by coacervation techniques or by interfacial polymerization, in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., ed. (1980). The particles or liposomes may further contain other therapeutic or diagnostic agents.

[0230] The precise therapeutic dosage and duration are a function of the disease being treated and, in some cases, can be determined empirically. Controlled clinical trials can also be conducted. The dosage can also vary with the severity of the condition to be alleviated. Pharmaceutical compositions are generally formulated and administered to exert a therapeutically applicable effect while minimizing undesirable side effects. The composition can be administered all at once or divided into multiple smaller doses administered at intervals. For any particular subject, the specific dosage regimen can be adjusted over time according to individual needs.

[0231] Thus, typical routes of administering these and related treatments or pharmaceutical compositions include, but are not limited to, oral, parenteral, nasal, intravenous, ophthalmic, intradermal, intramuscular, subcutaneous, instillation into the bladder, transdermal, inhalation, sublingual, buccal, rectal, vaginal, and topical. As used herein, the term parenteral includes subcutaneous injection, intravenous, instillation into the bladder, intramuscular, intrasternal injection, or infusion techniques. The treatment or pharmaceutical compositions according to certain embodiments of the present invention are formulated so that the active ingredients contained therein are bioavailable when the composition is administered to a subject or patient. The composition to be administered to a subject or patient can take the form of one or more dosage units, wherein, for example, a tablet can be a single dosage unit, and a container of the medicament described herein in aerosol form can accommodate multiple dosage units. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 23rd edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered will generally contain a therapeutically effective amount of an agent described herein for treating the disease or condition of interest.

[0232] The therapeutic or pharmaceutical composition may be in solid or liquid form. In some embodiments, the carrier is a microparticle, such that the composition is, for example, in tablet or powder form. The carrier may be a liquid, such that the composition is, for example, an oral oil, an injectable liquid, or an aerosol suitable for administration, such as by inhalation. When oral administration is desired, the pharmaceutical composition is preferably in solid or liquid form, with semi-solid, semi-liquid, suspension, and gel forms being included within the forms considered herein as solid or liquid. Certain embodiments include sterile injectable solutions.

[0233] As a solid composition for oral administration, the pharmaceutical composition can be formulated as a powder, granules, gel, compressed tablet, pill, capsule, chewable tablet, powder tablet, etc. The solid composition typically contains one or more inert diluents or edible carriers. In addition, one or more of the following may be present: a binder such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth gum or gelatin; an excipient such as starch, lactose or dextrin; a disintegrant such as alginic acid, sodium alginate, sodium starch glycolate, corn starch, etc.; a lubricant such as magnesium stearate or hydrogenated vegetable oil (Sterotex); a slip agent such as colloidal silicon dioxide; a sweetener such as sucrose or saccharin; a flavoring such as peppermint, methyl salicylate or citrus flavor; and a coloring agent. When the pharmaceutical composition is in the form of a capsule (e.g., a gelatin capsule), in addition to the above types of substances, it may contain a liquid carrier such as polyethylene glycol or oil.

[0234] The therapeutic or pharmaceutical composition can be in liquid form, such as an elixir, syrup, solution, gel, emulsion, or suspension. As two examples, the liquid can be administered orally or delivered by injection. When oral administration is desired, the preferred composition contains one or more of a sweetener, a preservative, a dye / colorant, and a flavor enhancer in addition to the compounds of the invention. In compositions intended for administration by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersant, a suspending agent, a buffer, a stabilizer, and an isotonic agent can be included.

[0235] Liquid therapeutic or pharmaceutical compositions, whether in the form of solutions, suspensions, or other similar solutions, may include one or more of the following adjuvants: sterile diluents such as water for injection, physiological saline solution (preferably physiological saline), Ringer's solution, isotonic sodium chloride, fixed oils (such as synthetic monoglycerides or diglycerides, which can serve as solvents or suspending media), polyethylene glycol, glycerol, propylene glycol, or other solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates, or phosphates, and agents for adjusting tonicity such as sodium chloride or dextrose. Parenteral formulations can be packaged in ampoules, disposable syringes, or multiple-dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. Injectable pharmaceutical compositions are preferably sterile.

[0236] Liquid therapeutic or pharmaceutical compositions intended for parenteral, intraocular, or oral administration should contain an amount of the agent that will achieve a suitable dosage. Typically, this amount is at least 0.01% of the relevant agent in the composition. When oral administration is desired, this amount may vary between 0.1% and about 70% of the composition weight. Certain oral therapeutic or pharmaceutical compositions contain between about 4% and about 75% of the agent of interest. In certain embodiments, therapeutic or pharmaceutical compositions and formulations are prepared so that a parenteral dosage unit contains between 0.01 and 10% by weight of the agent of interest prior to dilution.

[0237] The therapeutic or pharmaceutical composition may be intended for topical administration, in which case the formulation may suitably comprise a solution, emulsion, ointment, or gel base. For example, the base may comprise one or more of paraffin wax, wool wax, polyethylene glycol, beeswax, mineral oil, a diluent such as water and alcohol, and an emulsifier and stabilizer. A thickening agent may be present in the therapeutic or pharmaceutical composition for topical administration. If transdermal administration is desired, the composition may comprise a transdermal patch or iontophoresis device.

[0238] Therapeutic or pharmaceutical compositions may desirably be administered rectally, for example, in the form of suppositories that will melt in the rectum and release the drug. Compositions for rectal administration may contain an oily base as a suitable non-irritating excipient. Such bases include, but are not limited to, wool wax, cocoa butter, and polyethylene glycol.

[0239] Therapeutic or pharmaceutical compositions may include various materials that change the physical form of a solid or liquid dosage unit. For example, a composition may include a material that forms a coating shell around the active ingredient. The material forming the coating shell is typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be enclosed in a gelatin capsule. Therapeutic or pharmaceutical compositions in solid or liquid form may include components that bind to the agent and thereby help deliver the compound. Suitable components that may function in this capacity include monoclonal or polyclonal antibodies, one or more proteins, or liposomes.

[0240] Treatment or pharmaceutical composition can be basically made up of dosage units that can be used in aerosol form.The term aerosol is used to represent various systems, ranging from systems of colloidal nature to systems consisting of pressurized packages. Delivery can be carried out by liquefying or compressed gas or by a suitable pump system for distributing active ingredients. Aerosol can be delivered in the form of a single-phase, two-phase or three-phase system to deliver active ingredients. The delivery of aerosol includes necessary containers, activators, valves, secondary containers, etc., which together can form a test kit. Those of ordinary skill in the art can determine preferred aerosols without excessive experimentation.

[0241] The compositions described herein can be prepared with carriers that prevent rapid elimination of the agent from the body, such as time-release formulations or coatings. Such carriers include controlled-release formulations, such as, but not limited to, implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and other carriers known to those of ordinary skill in the art.

[0242] Pharmaceutical compositions can be prepared by methods well known in the pharmaceutical art. For example, a therapeutic or pharmaceutical composition intended for administration by injection may comprise one or more of a salt, a buffer, and / or a stabilizer, along with sterile distilled water to form a solution. A surfactant may be added to facilitate the formation of a homogeneous solution or suspension. A surfactant is a compound that non-covalently interacts with the pharmaceutical agent to facilitate dissolution or homogeneous suspension of the pharmaceutical agent in an aqueous delivery system.

[0243] The therapeutic or pharmaceutical composition can be administered in a therapeutically effective amount, which will vary depending on a variety of factors, including the activity of the specific compound employed; the metabolic stability and duration of action of the compound; the age, weight, general health, sex, and diet of the subject; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disorder or condition; and the subject undergoing therapy.

[0244] Also included are patient care kits comprising (a) an antibody that binds to IL-18BP as described herein; and optionally (b) at least one additional therapeutic agent. In some kits, (a) and (b) are in separate therapeutic compositions. In some kits, (a) and (b) are in the same therapeutic composition.

[0245] The kits herein may also include one or more additional therapeutic agents or other components suitable for or required for the indication being treated or to achieve the desired diagnostic application. The kits herein may also include one or more syringes or other components necessary or desired to facilitate the desired mode of delivery (e.g., stents, implantable reservoirs, etc.).

[0246] In some embodiments, the patient care kit contains separate containers, partitions, or compartments for the composition and informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material may be contained in association with the container. In some embodiments, the separate kit elements are contained in a single undivided container. For example, the composition is contained in a bottle, vial, or syringe to which informational material in the form of a label is attached. In some embodiments, the kit includes a plurality (e.g., a group) of individual containers, each container containing one or more unit dosage forms (e.g., dosage forms described herein) of an antibody and optionally at least one additional therapeutic agent. For example, the kit includes a plurality of syringes, ampoules, foil packets, or blister packs, each containing a single unit dose of an antibody and optionally at least one additional therapeutic agent. The container of the kit can be airtight, waterproof (e.g., watertight due to permeation or evaporation) and / or opaque.

[0247] The patient care kit optionally includes a device suitable for administering the composition, such as a syringe, an inhaler, a dropper (e.g., an eye dropper), a swab (e.g., a cotton swab or a wooden swab), or any such delivery device. In some embodiments, the device is an implantable device that dispenses a metered dose of the medicament. Also included are methods of providing a kit, for example, by combining the components described herein.

[0248] Expression and purification systems

[0249] Certain embodiments include methods and related compositions for expressing and purifying the anti-IL-18BP antibodies described herein. Such recombinant anti-IL-18BP antibodies are preferably prepared using standard protocols as described, for example, in Sambrook et al. (1989, supra), especially Chapters 16 and 17; Ausubel et al. (1994, supra), especially Chapters 10 and 16; and Coligan et al., Current Protocols in Protein Science (John Wiley & Sons, Inc. 1995-1997), especially Chapters 1, 5, and 6. As a general example, an anti-IL-18BP antibody can be prepared by a procedure comprising one or more of the following steps: (a) preparing a construct comprising a polynucleotide sequence encoding an anti-IL-18BP antibody heavy chain and / or light chain and operably linked to a regulatory element; (b) introducing the construct into a host cell; (c) culturing the host cell to express the anti-IL-18BP antibody; and (d) isolating the anti-IL-18BP from the host cell.

[0250] Thus, certain embodiments include polynucleotides encoding the anti-IL-18BP antibodies described herein, including vectors comprising these polynucleotides, and host cells comprising these polynucleotides and / or vectors. To express the desired polypeptide, the nucleotide sequence encoding the anti-IL-18BP or a functional equivalent thereof can be inserted into an appropriate expression vector (i.e., a vector containing the necessary elements for transcription and translation of the inserted coding sequence). Methods well known to those skilled in the art can be used to construct expression vectors containing sequences encoding the polypeptide of interest and appropriate transcription and translation control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described in Sambrook et al., Molecular Cloning: A Laboratory Manual (1989) and Ausubel et al., Current Protocols in Molecular Biology (1989).

[0251] A variety of expression vector / host systems are known and can be used to contain and express polynucleotide sequences. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., Cauliflower mosaic virus, CaMV; Tobacco mosaic virus, TMV) or bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems, including mammalian cells, and more particularly human cell systems.

[0252] "Control elements" or "regulatory sequences" present in an expression vector are those untranslated regions of the vector, i.e., enhancers, promoters, 5' and 3' untranslated regions, which interact with host cell proteins for transcription and translation. The strength and specificity of such elements can vary. Depending on the vector system and host utilized, any number of suitable transcription elements and translation elements can be used, including constitutive promoters and inducible promoters. For example, when cloning in a bacterial system, an inducible promoter can be used, such as the hybrid lacZ promoter of the PBLUESCRIPT phagemid (Stratagene, La Jolla, Calif.) or the PSPORT1 plasmid (Gibco BRL, Gaithersburg, Md.). In mammalian cell systems, promoters from mammalian genes or from mammalian viruses are generally preferred. If it is desired to produce a cell line containing multiple copies of a sequence encoding a polypeptide, then vectors based on SV40 or EBV should be used together with appropriate selectable markers.

[0253] In bacterial systems, a variety of expression vectors can be selected depending on the intended use of the expressed polypeptide. For example, when large quantities are required, vectors that direct high expression of fusion proteins that are easily purified can be used. Such vectors include, but are not limited to, versatile E. coli cloning and expression vectors, such as BLUESCRIPT (Stratagene), in which the sequence encoding the polypeptide of interest can be ligated in frame with the amino-terminal Met and the last seven residues of β-galactosidase to produce a hybrid protein; pIN vectors (Van Heeke and Schuster, J. Biol. Chem. 264:5503-5509 (1989)), etc. pGEX vectors (Promega, Madison, Wis.) can also be used to express exogenous polypeptides as fusion proteins with glutathione S-transferase (GST). Generally, such fusion proteins are soluble and can be easily purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems can be designed to include heparin, thrombin, or Factor XA protease cleavage sites, allowing for the free release of the cloned polypeptide of interest from the GST moiety.

[0254] Certain embodiments may employ an E. coli-based expression system (see, e.g., Structural Genomics Consortium et al., Nature Methods 5:135-146, 2008). These and related embodiments may rely, in part or in whole, on independent conjugation-dependent cloning (LIC) to create a suitable expression vector. In specific embodiments, protein expression may be controlled by T7 RNA polymerase (e.g., the pET vector series). These and related embodiments may utilize the expression host strain BL21 (DE3), a DE3 lysogen of BL21 that supports T7-mediated expression and lacks the lon and ompT proteases that improve the stability of the target protein. Also included are expression host strains carrying plasmids encoding tRNAs that are rarely used in E. coli, such as ROSETTA TM (DE3) and Rosetta 2 (DE3) strains. Cell lysis and sample processing can also be performed using the Nucleases and Protein extraction reagents are sold as improvements to the reagents. For cell culture, autoinduction media can improve the efficiency of many expression systems, including high-throughput expression systems. This type of media (e.g., OVERNIGHT EXPRESS TM Autoinduction systems) gradually induce protein expression via metabolic shift without the addition of artificial inducers (such as IPTG). Specific embodiments utilize a hexahistidine tag (such as the one labeled fusions) followed by immobilized metal affinity chromatography (IMAC) purification, or related techniques. However, in certain aspects, clinical-grade proteins can be isolated from E. coli inclusion bodies without or with affinity tags (see, e.g., Shimp et al., Protein Expr Purif 50:58-67, 2006). As another example, certain embodiments may employ a cold-shock-induced high-yield production system in E. coli, as overexpression of proteins in E. coli at low temperatures improves their solubility and stability (see, e.g., Qing et al., Nature Biotechnology 22:877-882, 2004).

[0255] High-density bacterial fermentation systems are also included. For example, high cell density culture of Ralstonia eutropha allows protein production at cell densities exceeding 150 g / L and expression of recombinant proteins at titers exceeding 10 g / L.

[0256] In the yeast Saccharomyces cerevisiae, a variety of vectors containing constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH can be used. For review, see Ausubel et al. (supra) and Grant et al., Methods Enzymol. 153:516-544 (1987). Also included are Pichia pandoris expression systems (see, for example, Li et al., Nature Biotechnology 24, 210-215, 2006; and Hamilton et al., Science 301:1244, 2003). Certain embodiments include yeast systems engineered to selectively glycosylate proteins, particularly including yeast with humanized N-glycosylation pathways (see, e.g., Hamilton et al., Science 313: 1441-1443, 2006; Wildt et al., Nature Reviews Microbiol. 3: 119-28, 2005; and Gerngross et al., Nature-Biotechnology 22: 1409-1414, 2004; U.S. Patent Nos. 7,629,163, 7,326,681, and 7,029,872). By way of example only, recombinant yeast cultures can be grown in Fernbach flasks or 15 L, 50 L, 100 L, and 200 L fermentors.

[0257] In the case of using a plant expression vector, the expression of the sequence encoding the polypeptide can be driven by any one of a variety of promoters. For example, viral promoters such as the 35S and 19S promoters of CaMV can be used alone or in combination with the ω leader sequence from TMV (Takamatsu, EMBO J. 6: 307-311 (1987)). Alternatively, plant promoters such as the small subunits of RUBISCO or heat shock promoters (Coruzzi et al., EMBO J. 3: 1671-1680 (1984) can be used; Broglie et al., Science (Science) 224: 838-843 (1984); and Winter et al., Results and Problems of Cell Differentiation (Results Probl. Cell Differ.) 17: 85-105 (1991)). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in several generally available reviews (see, eg, Hobbs, McGraw Hill, Yearbook of Science and Technology, pp. 191-196 (1992)).

[0258] Insect systems can also be used to express polypeptides of interest. For example, in one such system, the Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes in Spodopterafrugiperda cells or Trichoplusia cells. The sequence encoding the polypeptide can be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under the control of the polyhedrin promoter. Successful insertion of the polypeptide coding sequence will render the polyhedrin gene inactive and produce a recombinant virus lacking sheath proteins. The recombinant virus can then be used to infect, for example, Spodopterafrugiperda cells or Trichoplusia cells that can express the polypeptide of interest (Engelhard et al., Proc. Natl. Acad. Sci. USA 91: 3224-3227 (1994)). Also included are baculovirus expression systems, including those utilizing SF9, SF21, and Tni cells (see, e.g., Murphy and Piwnica-Worms, Curr Protoc Protein Sci. Chapter 5: Unit 5.4, 2001). Insect systems can provide similar post-translational modifications as mammalian systems.

[0259] In mammalian host cells, a variety of viral-based expression systems are generally available. For example, when adenovirus is used as an expression vector, the sequence encoding the polypeptide of interest can be joined to an adenoviral transcription / translation complex consisting of a late promoter and a tripartite leader sequence. Insertion into the non-essential E1 or E3 region of the viral genome can be used to obtain a live virus capable of expressing polypeptides in infected host cells (Logan and Shenk, Proc. Natl. Acad. Sci. USA 81: 3655-3659 (1984)). In addition, transcription enhancers, such as Rous sarcoma virus (RSV) enhancers, can be used to increase expression in mammalian host cells.

[0260] Examples of suitable mammalian host cell lines include monkey kidney CV1 strain transformed with SV40 (COS-7, ATCC CRL1651); human embryonic kidney strain (293 cells or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10); mouse sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human hepatocytes (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TR1 cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and human liver tumor line (Hep G2). Other suitable mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR-CHO cells (Urlaub et al., PNAS USA 77:4216 (1980)); and myeloma cell lines, such as NSO and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, for example, Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (BKC Lo, ed., Humana Press, Totowa, NJ, 2003), pp. 255-268. Certain preferred mammalian cell expression systems include those based on CHO and HEK293 cells. Mammalian expression systems can utilize, among others, adherent cell lines in, for example, T flasks, roller bottles, or cell factories; or suspension cultures in, for example, 1 L and 5 L spinners, 5 L, 14 L, 40 L, 100 L, and 200 L stirred tank bioreactors, or 20 / 50 L and 100 / 200 L WAVE bioreactors, among those known in the art.

[0261] Cell-free expression of proteins is also included.These and related embodiments generally utilize purified RNA polymerase, ribosomes, tRNA, and ribonucleotides; these reagents can be produced by extraction from cells or from cell-based expression systems.

[0262] In some embodiments, the initiation signal of the present invention can be used to realize the more effective translation of the sequence of the polypeptide of interest. Such signals include ATG start codon and adjacent sequences. When the sequence of the coded polypeptide, its start codon and upstream sequence are inserted into an appropriate expression vector, additional transcription or translation control signals may not be needed. However, when inserting only the coding sequence or a part thereof, an exogenous translation control signal including the ATG start codon should be provided. In addition, the start codon should be in the correct reading frame to ensure the translation of the entire insert. Exogenous translation elements and start codons can be derived from various sources (natural and synthetic sources). Expression efficiency can be enhanced by including an enhancer suitable for the specific cell system used, such as those described in the literature (Scharf et al., Results and Problems of Cell Differentiation (Results Probl. Cell Differ.) 20: 125-162 (1994)).

[0263] In addition, host cell strains can be selected for the expression of its regulated insertion sequence or the ability of processing expressed protein in a desired manner. This type of polypeptide modification includes but is not limited to post-translational modification, such as acetylation, carboxylation, glycosylation, phosphorylation, lipidation and acylation. The post-translational processing of the "pre-form" form of cleaved protein can also be used to facilitate correct insertion, folding and / or function. Except bacterial cells, can select to have or even lack the different host cells of active specific cell mechanism and characteristic mechanism at this type of post-translational, such as yeast, CHO, HeLa, MDCK, HEK293 and W138, to ensure correct modification and processing of foreign protein.

[0264] For producing recombinant protein in high yield for a long time, stable expression is generally preferred. For example, the cell line of the polynucleotide of stable expression can be transformed using expression vectors, and these expression vectors can contain viral replication origin and / or endogenous expression element and selectable marker gene on the same or separate carrier. After introducing the vector, the cell can be grown in enrichment medium for about 1-2 days and then switched to selective medium. The purpose of the selectable marker is to give resistance to selection, and its presence allows growth and recovery of the cell of the successfully expressed sequence. The resistant clone of the tissue culture technique that is suitable for the cell type can be used to proliferate stably transformed cells. It is also possible to adopt short-term production, such as short-term transfection or infection. The exemplary mammalian expression system that is suitable for short-term production includes the system based on HEK293 and CHO.

[0265] Any number of selection systems can be used to reclaim transformed or transduced cell lines. These selection systems include, but are not limited to, herpes simplex virus thymidine kinase (Wigler et al., Cell 11: 223-232 (1977)) and adenine phosphoribosyltransferase (Lowy et al., Cell 22: 817-823 (1990)) genes, which can be used for tk-cells or aprt-cells, respectively. In addition, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection; for example, dhfr, which confers resistance to methotrexate (Wigler et al., PNAS USA. 77:3567-70 (1980)); npt, which confers resistance to aminoglycosides, neomycin, and G-418 (Colbere-Garapin et al., J. Mol. Biol. 150:1-14 (1981)); and als or pat, which confer resistance to chlorsulfuron and phosphinothricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, such as trpB, which allows cells to utilize indole instead of tryptophan, or hisD, which allows cells to utilize histadinol instead of histidine (Hartma and Mulligan, Proc. Natl. Acad. Sci. USA 85:8047-51 (1988)). The use of visible markers has become widespread, and such markers, such as green fluorescent protein (GFP) and other fluorescent proteins (e.g., RFP, YFP), anthocyanins, β-glucuronidase and its substrate GUS, and luciferase and its substrate luciferin, are widely used not only to identify transformants, but also to quantify the amount of transient or stable protein expression caused by a particular vector system (see, e.g., Rhodes et al., Methods Mol. Biol. 55: 121-131 (1995)).

[0266] Also included are high-throughput protein production systems or microproduction systems. In certain aspects, for example, a hexa-histidine fusion tag can be used for protein expression and purification on a slide surface modified with a metal chelator or MagneHis Ni particles (see, e.g., Kwon et al., BMC Biotechnol. 9:72, 2009; and Lin et al., Methods Mol Biol. 498:129-41, 2009). Also included are high-throughput cell-free protein expression systems (see, e.g., Sitaraman et al., Methods Mol Biol. 498:229-44, 2009). These and related embodiments can be used, for example, to generate microarrays of antibodies, which can then be used to screen libraries to identify antibodies and antigen-binding domains that interact with the IL-18BP polypeptide of interest.

[0267] Although the foregoing embodiments have been described in considerable detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to one of ordinary skill in the art in light of the teachings of the present invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. The following examples are provided by way of illustration only and not by way of limitation. One of ordinary skill in the art will readily recognize a variety of non-critical parameters that may be changed or modified to produce substantially similar results.

[0268] Illustrative Examples

[0269] The following non-limiting listed examples are provided by way of illustration.

[0270] Example I-1. An antibody specific for interleukin-18 binding protein (IL-18BP), wherein the antibody interferes with the binding of IL-18 to IL-18BP.

[0271] Example I-2. The antibody of Example I-1, wherein the antibody binds to a conformational epitope of IL-18BP.

[0272] Embodiment 1-3. The antibody of embodiment 1-2, wherein the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO: 372.

[0273] Embodiment 1-4. The antibody of embodiment 1-2, wherein the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116 and S119 of SEQ ID NO: 372.

[0274] Embodiment I-5. The antibody of embodiment I-1, wherein the antibody binds to a linear epitope of IL-18BP.

[0275] Embodiment I-6. The antibody of any one of embodiments I-1 to I-5, wherein the antibody binds to the binding interface between IL-18 and the mature form of IL-18BP.

[0276] Example 1-7. The antibody of Example 1-6, wherein the antibody binds to amino acid residues S75, H79, T116, and S119 of SEQ ID NO: 372.

[0277] Embodiment 1-8. The antibody of any one of embodiments 1-7, wherein the antibody binds to human IL-18BP and cynomolgus monkey IL-18BP, but does not bind to mouse IL-18BP.

[0278] Embodiment I-9. The antibody of any one of Embodiments I- to I-7, wherein the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

[0279] Embodiment I-10. The antibody of any one of embodiments I-1 to I-9, wherein the antibody binds to IL-18 with a binding affinity (K) greater than that between IL-18 and IL-18BP. D 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM), optionally binds to IL-18BP with a binding affinity of about 1 pM to about 650 pM, or about or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM.

[0280] Embodiment I-11. The antibody of any one of Embodiments I-1 to I-10, wherein the antibody is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18.

[0281] Embodiment I-12. The antibody of embodiment I-11, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18 and thereby increases IL-18-mediated signaling, including the induction of IFN-γ, CXCL10 and / or TNFα.

[0282] Embodiment I-13. The antibody of any one of embodiments I-1 to I-12, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3 and IgG4) or IgM Fc domain, optionally a human Fc domain or a hybrid and / or variant thereof.

[0283] Embodiment I-14. The antibody of embodiment I-13, comprising an IgG Fc domain with high effector function in humans, optionally an IgG1 or IgG3 Fc domain.

[0284] Example I-15. The antibody of Example I-13, comprising an IgG Fc domain with low effector function in humans, optionally an IgG2 or IgG4 Fc domain.

[0285] Embodiment I-16. The antibody of any one of embodiments I-1 to I-15, wherein the antibody is a monoclonal antibody.

[0286] Embodiment I-17. The antibody of any one of embodiments I-1 to I-16, wherein the antibody is a humanized antibody.

[0287] Embodiment I-18. The antibody of any one of Embodiments I-1 to I-17, wherein the antibody is a scFv.

[0288] Embodiment I-19. The antibody of any one of embodiments I-1 to I-18, wherein the antibody comprises:

[0289] a) Heavy chain variable region (V H ), which comprises a complementarity determining region V selected from Table A1 H CDR1, V H CDR2 and V H CDR3 sequences and variants thereof that specifically bind to IL-18BP; and

[0290] b) Light chain variable region (V L ), which comprises a complementarity determining region V selected from Table A1 L CDR1, V L CDR2 and V L CDR3 sequences and their variants that specifically bind to IL-18BP.

[0291] Embodiment 1-20. The antibody of embodiment 1-19, wherein:

[0292] a) the V H CDR1, V H CDR2 and V HThe CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 4-6, respectively;

[0293] b) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 7-9, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 10-12, respectively;

[0294] c) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 13-15, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 16-18, respectively;

[0295] d) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 19-21, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 22-24, respectively;

[0296] e) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 28-30, respectively;

[0297] f) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 31-33, respectively, and the V L CDR1, V LCDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 34-36, respectively;

[0298] g) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 37-39, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 40-42, respectively;

[0299] h) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 46-48, respectively;

[0300] i) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 49-51, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 52-54, respectively;

[0301] j) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 58-60, respectively;

[0302] k) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 64-66, respectively;

[0303] 1) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 70-72, respectively;

[0304] m) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 73-75, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 76-78, respectively;

[0305] n) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 79-81, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 82-84, respectively;

[0306] o) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 85-87, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 88-90, respectively;

[0307] p) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 91-93, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 94-96, respectively;

[0308] q) V H CDR1, V H CDR2 and VH The CDR3 sequences comprise SEQ ID NOs: 97-99, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 100-102, respectively;

[0309] r) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 103-105, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 106-108, respectively;

[0310] s) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 112-114, respectively;

[0311] t) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 118-120, respectively;

[0312] u) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 124-126, respectively;

[0313] v) V H CDR1, V H CDR2 and V HThe CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 130-132, respectively;

[0314] w) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 136-138, respectively;

[0315] x) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 142-144, respectively;

[0316] y) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 148-150, respectively;

[0317] z) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 154-156, respectively;

[0318] aa) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and the VL CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 160-162, respectively;

[0319] bb) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 166-168, respectively;

[0320] cc) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 172-174, respectively;

[0321] dd) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 178-180, respectively;

[0322] ee) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 184-186, respectively;

[0323] ff) the V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and the V L CDR1, V LCDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 190-192, respectively;

[0324] gg) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 193-195, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 196-198, respectively;

[0325] hh) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 199-201, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 202-204, respectively;

[0326] ii) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 205-207, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 208-210, respectively;

[0327] jj) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 211-213, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 214-216, respectively;

[0328] kk) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 217-219, respectively, and the V L CDR1, V L CDR2 and V LThe CDR3 sequences comprise SEQ ID NOs: 220-222, respectively;

[0329] 11) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 223-225, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 226-228, respectively;

[0330] mm) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 229-231, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 232-234, respectively;

[0331] nn) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 235-237, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 238-240, respectively;

[0332] oo) said V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 241-243, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 244-246, respectively;

[0333] pp) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 250-252, respectively;

[0334] qq)V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 253-255, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 256-258, respectively; or

[0335] rr) V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 259-261, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 262-264, respectively.

[0336] Embodiment 1-21. The antibody of any one of embodiments 1-20, wherein the VH comprises a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein the V H There are 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in the framework regions.

[0337] Embodiment I-22. The antibody of any one of embodiments I-1 to I-21, wherein the VL comprises a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein the V L There are 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in the framework regions.

[0338] Embodiment 1-23. The antibody of any one of embodiments 1-1 to 1-22, wherein:

[0339] a) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 265, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 266;

[0340] b) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 267, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 268;

[0341] c) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 269, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 270;

[0342] d) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 271, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 272;

[0343] e) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 273, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 274;

[0344] f) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 275, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 276;

[0345] g) the V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 277, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 278;

[0346] h) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 279, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 280;

[0347] i) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 281, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 282;

[0348] j) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 283, and wherein said V L comprising a sequence at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 284;

[0349] k) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 285, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 286;

[0350] 1) the V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 287, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 288;

[0351] m) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 289, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 290;

[0352] n) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 291, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 292;

[0353] o) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 293, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 294;

[0354] p) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 295, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 296;

[0355] q) V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 297, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 298;

[0356] r) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 299, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 300;

[0357] s) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 301, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 302;

[0358] t) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 303, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 304;

[0359] u) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 305, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 306;

[0360] v) V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 307, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308;

[0361] w) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 309, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 310;

[0362] x) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 311, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 312;

[0363] y) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 313, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 314;

[0364] z) the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 315, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 316;

[0365] aa) V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 317, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 318;

[0366] bb) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 319, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 320;

[0367] cc) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 321, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 322;

[0368] dd) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 323, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 324;

[0369] ee) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 325, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 326;

[0370] ff) the V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 327, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 328;

[0371] gg) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 329, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 330;

[0372] hh) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 331, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 332;

[0373] ii) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 333, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 334;

[0374] jj) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 335, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 336;

[0375] kk) V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 337, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 338;

[0376] 11) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 339, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 340;

[0377] mm) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 341, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 342;

[0378] nn) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 343, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 344;

[0379] oo) said V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 345, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 346;

[0380] pp) V Hcomprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 347, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 348;

[0381] qq)V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 349, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 350; or

[0382] rr) V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 351, and wherein said V L Comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO:352.

[0383] Embodiment 1-24. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody of any one of Embodiments 1-1 to 1-23; an expression vector comprising the isolated polynucleotide; or an isolated host cell comprising the vector.

[0384] Embodiment I-25. A pharmaceutical composition comprising the anti-IL-18BP antibody of any one of Embodiments I-1 to I-23 and a pharmaceutically acceptable carrier.

[0385] Embodiment 1-26. The pharmaceutical composition of embodiment 1-25, wherein the composition is a sterile injectable solution optionally suitable for intravenous, intramuscular, subcutaneous or intraperitoneal administration.

[0386] Embodiment 1-27. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of Embodiments 1-25 to 1-26.

[0387] Embodiment 1-28. The method of embodiment 1-27, wherein the disease or condition is cancer or a tumor or a proliferative disease or disorder, optionally selected from the group consisting of a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enteritis, a proliferative diabetic retinopathy, and a proliferative nephropathy.

[0388] Embodiment 1-29. The method of embodiment 1-28, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or wherein the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

[0389] Embodiment 1-30. The method of embodiment 1-28 or 1-29, wherein the cancer is selected from one or more of the following: bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma), lymphoma), Hodgkin's lymphoma), liver cancer (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, urothelial cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and stomach cancer.

[0390] Embodiment I-31. The method of any one of embodiments I-27 to I-30, comprising administering the pharmaceutical composition of embodiment 25 or 26 in combination with IL-18.

[0391] Embodiment I-32. A method as described in any one of Embodiments I-28 to I-31, which comprises administering a pharmaceutical composition as described in Embodiment I-25 or I-26 in combination with an immune checkpoint regulator, wherein the immune checkpoint regulator is selected from antagonists of inhibitory immune checkpoint molecules and agonists of stimulatory immune checkpoint molecules.

[0392] Example 1-33. The method of Example 1-32, wherein the immune checkpoint regulator is a polypeptide, optionally an antibody or a ligand or a small molecule.

[0393] Example 1-34. A method as described in Example 1-32 or 1-33, wherein the inhibitory immune checkpoint molecule is selected from one or more of the following: programmed death-ligand 1 (PD-L1), programmed death 1 (PD-1), programmed death-ligand 2 (PD-L2), cytotoxic T lymphocyte-associated protein 4 (CTLA-4), indoleamine 2,3-dioxygenase (IDO), tryptophan 2,3-dioxygenase (TDO), T cell immunoglobulin domain and mucin domain 3 (TIM-3), lymphocyte activation gene-3 (LAG-3), V-domain Ig inhibitory factor of T cell activation (VISTA), B and T lymphocyte attenuator (BTLA), CD160, herpes virus entry mediator (HVEM) and T cell immunoreceptor with Ig and ITIM domains (TIGIT).

[0394] Embodiment 1-35. The method of embodiment 1-34, wherein:

[0395] a) the antagonist is a PD-L1 and / or PD-L2 antagonist, optionally selected from one or more of the following: an antibody or small molecule that specifically binds to PD-L1 and / or PD-L2, atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), optionally wherein the cancer is selected from one or more of the following: colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer, and renal cell carcinoma;

[0396] b) the antagonist is a PD-1 antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally wherein the PD-1 antagonist is nivolumab, and the cancer is optionally selected from one or more of the following: Hodgkin's lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer;

[0397] c) the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of the following: melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial carcinoma;

[0398] d) the antagonist is a CTLA-4 antagonist, optionally selected from one or more of the following: an antibody or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, optionally wherein the cancer is selected from one or more of the following: melanoma, prostate cancer, lung cancer, and bladder cancer;

[0399] e) the antagonist is an IDO antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-tryptophan (1MT), a β-carboline (norharmane; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, and wherein the cancer is optionally selected from one or more of the following: metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or a malignant brain tumor;

[0400] f) the antagonist is a TDO antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to TDO, 680C91, and LM10;

[0401] g) the antagonist is a TIM-3 antagonist optionally selected from one or more antibodies or small molecules that specifically bind to TIM-3;

[0402] h) the antagonist is a LAG-3 antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to LAG-3, and BMS-986016;

[0403] i) the antagonist is a VISTA antagonist optionally selected from one or more antibodies or small molecules that specifically bind to VISTA;

[0404] j) the antagonist is a BTLA, CD160 and / or HVEM antagonist optionally selected from one or more antibodies or small molecules that specifically bind to BTLA, CD160 and / or HVEM; or

[0405] k) The antagonist is a TIGIT antagonist, optionally selected from one or more antibodies or small molecules that specifically bind to TIGIT.

[0406] Example 1-36. A method as described in Example 1-32 or 1-33, wherein the stimulatory immune checkpoint molecule is selected from one or more of the following: OX40, CD40, glucocorticoid-induced TNFR family-related gene (GITR), CD137 (4-1BB), CD27, CD28, CD226 and herpes virus entry mediator (HVEM).

[0407] Embodiment 1-37. The method of embodiment 1-36, wherein:

[0408] a) the agonist is an OX40 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L and GSK3174998;

[0409] b) the agonist is a CD40 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, and rhCD40L, and wherein the cancer is optionally selected from one or more of the following: melanoma, pancreatic cancer, mesothelioma, and a blood cancer, optionally a lymphoma, such as non-Hodgkin's lymphoma;

[0410] c) the agonist is a GITR agonist optionally selected from one or more of the following: an antibody or a small molecule or a ligand that specifically binds to GITR, INCAGN01876, DTA-1, and MEDI1873;

[0411] d) the agonist is a CD137 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD137, utomilumab, and a 4-1BB ligand;

[0412] e) the agonist is a CD27 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5);

[0413] f) the agonist is a CD28 agonist optionally selected from one or more of the following: an antibody or a small molecule or a ligand that specifically binds to CD28, and TAB08; and / or

[0414] g) The agonist is an HVEM agonist optionally selected from one or more antibodies or small molecules or ligands that specifically bind to HVEM.

[0415] Embodiment 1-38. The method of any one of embodiments 1-28 to 1-37, comprising administering the pharmaceutical composition of embodiment 1-25 or 1-26 in combination with at least one chemotherapeutic agent.

[0416] Embodiment 1-39. The method of embodiment 1-38, wherein the at least one chemotherapeutic agent is one or more selected from the group consisting of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (Type 1 or Type II), and an antimicrotubule agent.

[0417] Embodiment 1-40. The method of embodiment 1-39, wherein:

[0418] a) the alkylating agent is one or more selected from the group consisting of nitrogen mustards (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosoureas (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine (fotemustine), and methylprednisolone. e) and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide and temozolomide), aziridines (optionally thiotepa, mytomycin and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaliplatin) and atypical alkylating agents (optionally procarbazine and hexamethonium);

[0419] b) the antimetabolite is one or more selected from the group consisting of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine);

[0420] c) the cytotoxic antibiotic is one or more selected from the group consisting of anthracyclines (optionally, doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin and mitoxantrone), bleomycin, mitomycin C, mitoxantrone and actinomycin;

[0421] d) the topoisomerase inhibitor is one or more selected from the group consisting of camptothecin, irinotecan, topotecan, etoposide, cypermethrin, mitoxantrone, teniposide, novobiocin, merbarone and aclarubicin; and / or

[0422] e) The anti-microtubule agent is one or more selected from the group consisting of taxanes (optionally, paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

[0423] Embodiment 1-41. The method of embodiment 1-27, wherein the disease or condition is myelodysplastic syndrome (MDS).

[0424] Embodiment 1-42. The method of embodiment 1-27, wherein the disease or condition is an infectious disease.

[0425] Embodiment 1-43. The method of embodiment 1-42, wherein the infectious disease is selected from viral, bacterial, fungal (optionally yeast) and protozoal infections.

[0426] Embodiment I-44. The method of any one of embodiments I-41 to I-43, comprising administering the pharmaceutical composition of embodiment I-25 or I-26 in combination with IL-18.

[0427] Example 1-45. A method for screening anti-IL-18BP antibodies for their ability to block or inhibit the binding between IL-18 and IL-18BP, comprising

[0428] a) determining the binding affinity of the antibody to (i) IL-18BP alone and (ii) a low IL-18 fusion protein, wherein the low IL-18 fusion protein comprises IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein binds to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein;

[0429] b) comparing the binding affinity of (i) to the binding affinity of (ii); and

[0430] c) If the binding affinity of (i) is significantly stronger than the binding affinity of (ii), then the antibody is identified or selected as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP.

[0431] Embodiment 1-46. The method of embodiment 1-45, wherein the IL-18 and the IL-18BP are mouse IL-18 and mouse IL-18BP.

[0432] Embodiment 1-47. The method of embodiment 1-45, wherein the IL-18 and the IL-18BP are human IL-18 and human IL-18BP.

[0433] Examples

[0434] Example 1: Generation of antagonistic monoclonal antibodies against interleukin-18 binding protein (IL-18BP)

[0435] Studies are underway to generate potent human and humanized antibodies that bind to and inhibit IL-18 / IL-18BP binding, thereby releasing IL-18 to stimulate immune activity. Potential therapeutic candidates with cross-reactivity to human and cynomolgus macaque IL-18BP are identified. Furthermore, potent anti-mouse IL-18BP mAbs closely related to their anti-human / cynomolgus macaque counterparts are generated to allow the study of these agents in murine tumor model systems.

[0436] Materials and methods

[0437] Immunization and isolation of antigen-specific antibodies from single B cells.

[0438] Monoclonal antibodies to IL-18BP are generated by immunizing mice with IL-18BP, followed by isolation of antigen-specific single B cells using a Berkeley Lights Beacon instrument and cloning of the antibody-encoding gene from each cell of interest. TM and DivergimAb TM (Abveris Inc.) was used to generate murine mAbs. Mice were immunized with alternating courses of human IL-18BP and cynomolgus macaque IL-18BP using either a his or Fc tag, while titers were tested with human, cynomolgus macaque, or mouse IL-18BP using the other tag in each case to prevent detection of tag-specific antibodies.

[0439] After generating high titers for IL-18BP, antigen-specific B cells from appropriate mice were isolated as single cells using a Berkeley Lights Beacon instrument as described using the manufacturer's recommended procedures (Mullen et al., Antibody Therapeutics 4(3):185-196, 2021). Immunoglobulin gene sequences from antigen-specific B cells were obtained and used to generate recombinant antibodies using established methods.

[0440] Name the antibody.

[0441] Some antibodies are designated with an "SA" prefix, a consecutive 2-digit number, and a single-letter suffix indicating the nature of the HC constant domain: "a" for human IgG1, "d" for murine IgG2a. Thus, SA04a is a human IgG1 antibody.

[0442] Variants and mutations are named in the following order: the original amino acid, followed by the Kabat position number (Kabat 1991), followed by the replacement amino acid. For amino acids, the standard single-letter code is used. Thus, Y32E indicates that the tyrosine (Y) at Kabat position 32 has been replaced by glutamic acid (E).

[0443] Library preparation / site-directed mutagenesis.

[0444] To affinity-mature the starting mAb, a library of variants was prepared, focusing sequentially on each of the HC and LC CDRs. To achieve this, each CDR amino acid was sequentially replaced with up to 17 amino acid substitutions. Cysteine ​​and tryptophan were excluded from the library to avoid introducing undesirable potential sequence liabilities, and the parent amino acid was not already present in the positions included in the screen. To generate variants at each position, two sets of mutagenic oligonucleotides (Integrated DNA Technologies (IDT), San Diego) containing the degenerate codons NDT or VHG (where N = A / C / G / T; D = A / G / T, V = A / C / G; H = A / C / T) were used for each position (Kille, 2013; Acevedo-Rocha, 2015), paired with appropriate 5' and 3' distal oligonucleotides (IDT) designed to permit amplification and cloning.

[0445] According to the manufacturer's protocol, PCR was performed with high-fidelity DNA polymerase (Q5, New England Biolabs). Parental plasmid (heavy chain and light chain) was diluted to 10 ng / μ L, and 1 μ L was used as the template for each 50 μ L reaction. The V region gene fragment with degenerate codons as described above was amplified by PCR, and purified (using Qiagen PCR purification kit according to manufacturer's instructions). Gene fragments were assembled into heavy and light chain clones via overlap extension PCR (OE-PCR) or Gibson cloning. For OE-PCR, the fragment was amplified with the corresponding forward and reverse primers containing restriction sites (AgeI-NheI for heavy chain, SbfI-MfeI for light chain) and column purified (Qiagen PCR purification kit). Restriction digestion was performed using high-fidelity enzyme (New England Biolabs), and the fragment was joined to the appropriate vector for heavy and light chain using T4 DNA ligase (New England Biolabs, catalog number (Cat. No.) M0202L). The empty heavy chain vector contains most of the human IgG1 constant region, with an engineered NheI site (created by changing the wobble position) of 12 amino acids introduced into the constant region. The empty light chain vector contains most of the human kappa constant region, with an engineered MfeI site of 18 amino acids introduced into the constant region. Gibson cloning (Gibson Master mix kit, New England Biolabs catalog number ES2611L). Inserts were normalized to 1 ng / μL and a total of 2 ng of insert DNA (1 ng per fragment) was used. A 10 μL reaction volume was made up of 5 μL of Gibson master mix and QS with purified water. Reactions were incubated at 50°C for 15-60 minutes.

[0446] Ligation products from OE-PCR or Gibson assembly were transformed into competent E. coli (Monserate Biotechnology, San Diego) by adding 2-5 μL of ligation reaction mixture to the cells and incubating on ice for 5 minutes. The cells were heat shocked at 42°C for 30 seconds and placed on ice. 250 μL of SOC medium (BioPioneer, San Diego or Teknova, San Diego) was added and the tubes were incubated at 37°C, 200 rpm for 1 hour. 100 μL of culture was plated onto antibiotic selection plates (BioPioneer, San Diego or Teknova, San Diego) and incubated overnight at 37°C.

[0447] The delivery tray is to carry out colony sequencing (Genewiz or Eton) to the antibody gene, and wherein each culture plate sequencing clone is 24-48.Sequence is analyzed for reference sequence with SnapGene software (GSL Biotech), and described reference sequence is the computer simulation clone of library.Based on sequence alignment and the amino acid encoded by mutation primer, clone is selected, and these clones are used to produce individual plasmid mini-preps.

[0448] Screening of mutants was achieved using plasmids derived from a small-scale expression library in Expi293F cells cultured in 48-well plates using the method described below. Heavy and light chain pairing was performed at a 1:2 ratio (0.5 ng HC: 1 ng LC plasmid / well). A replicate of the parental antibody control was included on each plate. Cells were grown in the plates for 3 days, after which supernatants from each well were collected for screening.

[0449] Screening was performed using biofilm interferometry (BLI) for plate transfection. Initially, BLI was used to determine the concentration of antibody present in each sample before screening for binding affinity to IL-18BP. Initial apparent binding kinetic measurements were obtained on a Fortébio Octet RED96e instrument. mAbs were loaded onto anti-human constant domain (AHC) biosensors (FortéBio) in 10× kinetic buffer consisting of PBS, 0.1% BSA, and 0.02% Tween 20 for 120 seconds to achieve a spectral shift value of 0.8 to 1.2 nm. The association phase was performed in the presence of 20 nM human, cynomolgus macaque, or murine IL-18BP orthologs and allowed to proceed for 120 seconds; dissociation was measured for 300 seconds to determine whether any variants exhibited an increased on- or off-rate compared to the parent antibody. Each ortholog was then compared for full binding kinetics, and candidates that exhibited significantly improved binding kinetics based on a single screening concentration were retested and recombined with additional mutations as described below.

[0450] Expression and purification of recombinant antibodies.

[0451] Grow Expi293F cells from the Expi293 Expression System Kit (Thermo Fisher, catalog number A14635) in Expi293F Expression Medium (catalog number A1435101). Grow cells to 3-6×10 6The cells were cultured at a density of 10 cells / mL and then counted using a hemocytometer. Plasmid DNA (1.0 μg / 1.0 mL culture) was diluted in Opti-MEM Reduced Serum Medium (RSM) (Cat. No. 31985062). The values ​​for Opti-MEM RSM were obtained according to the manufacturer's recommendations for transfection. Expifectamine 293 reagent was diluted in Opti-MEM RSM and incubated at room temperature for 5 minutes before mixing with the diluted plasmid DNA. This mixture was allowed to stand and incubate at room temperature for 10-20 minutes. While incubating the expifectamine / plasmid DNA complex, Expi293F cells were diluted to 3×10 6 Cells were transfected at a density of 10 cells / mL and added to the desired volume in an Erlenmeyer flask (BioPioneer, DGFPC0125S for 125 mL flasks). The expifectamine / plasmid DNA complex was then slowly transferred to a shake flask containing Expi293F cells, and the flask was placed in a shaking incubator (Infors-HT Multitron) at 37°C, 8% CO2, and 125 rpm with a 25 mm orbital throw. 18-22 hours post-transfection, ExpiFectamine 293 transfection enhancers 1 (#100013863) and 2 (#A14350-01) were added to the cells, and the cells were returned to the shaking incubator. The cells were then incubated for an additional 4 days, followed by rapid centrifugation at 4000 × g for 20 minutes in a refrigerated centrifuge and filtration at 0.22 μm prior to purification.

[0452] On AKTAExplorer FPLC system, use 5mL HiTrap MabSelect SuRe (protein A) column purification antibody.First, by adding 50mL 0.1M glycine, pH 3.0 (elution buffer), add 50mL 50mM glycine, 50mM glycinate pH 8 (binding / wash buffer) subsequently and remove any residual bound protein in the post. With 5mL / min, antibody (25-400mL) is loaded on the post, and further washes with 25mL balance / wash buffer, until UV reading reaches baseline. Subsequently, by using the 25mL linear gradient of 0-100% elution buffer with 5mL / min wash-out MAb 2min. According to 280nm absorbance monitoring antibody elution. Collect the peak elution fraction and pool in the 15mL conical tube. The material was then buffer exchanged into storage buffer (PBS, pH 7.4) using PD10 columns (Cytiva Cat. No. 17085101) and then filter sterilized (GenClone syringe filters, Cat. No. 25-244, attached to BD 5 mL [Cat. No. 309646] and 20 mL [Cat. No. 302830] BD Luer-Lok TM syringe) into a 15 mL conical tube and used for subsequent characterization analysis.

[0453] Analysis by size exclusion HPLC (SEC-HPLC).

[0454] SEC-HPLC was performed on a TSKgel G3000SW with a particle size of 5 μm and a 7.8 mm ID × 30 cm XL The HPLC was performed on an Agilent 1100 HPLC column and run isocratically at a flow rate of 1 mL / min using 50 mM sodium phosphate, 200 mM arginine, pH 6.8. Detection was performed at 280 nm using a diode array detector and peaks were integrated using Agilent ChemStation software. The SEC-HPLC standards used to calibrate the column consisted of bovine thyroglobulin, bovine IgG, chicken albumin, bovine ribonuclease A, and p-aminobenzoic acid (Sigma Aldrich #69385).

[0455] The binding affinity was analyzed by biomembrane interferometry (BLI).

[0456] Binding kinetic measurements were obtained on a Fortébio (now Sartorius) Octet RED96e instrument. mAbs were loaded onto anti-human constant domain (AHC) biosensors (FortéBio) in 10× kinetic buffer consisting of PBS containing 0.1% BSA and 0.02% Tween 20 for 90–120 s to achieve spectral shifts of 0.8 to 1.2 nm. Association was performed in the presence of a two-fold dilution series of hIL-18BP and typically proceeded for 90–120 s; dissociation was typically measured for 300 to 1200 s. Dilution series began at 100 nM for weaker variants or 10 nM for the most potent mAbs. Cross-reactivity to cynomolgus monkey IL-18BP and mouse IL-18BP was determined using the same method as for IL-18BP from the appropriate species.

[0457] IL-18 reporter activity of anti-IL-18BP mAb in HEK 293 cells.

[0458] IL-18 reporter HEK 293 cells (hkb-hmil18) from InvivoGen respond to exogenously added IL-18 by expressing an NF-κB / AP-1 inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene. For analysis, cells were grown in complete DMEM medium (10% HI FBS, 1% PS) with 1× HEK-Blue Selection (Invivogen, hb-sel). Cells were carefully rinsed twice with 1× PBS and incubated with 1× PBS at 37°C for 5 minutes. Cells were counted, centrifuged quickly at 200×g for 5 minutes, and centrifuged at 2.5×10 5The cells were then resuspended in complete DMEM medium without selection. The cells were then plated at 25,000 cells / mL in 100 μL / well in a 96-well plate (Genesee Scientific, 25-109). The plated cells were then placed in an incubator at 37°C, 5% CO2. A test antibody solution was prepared at 90 μg / mL in DMEM without selection and incubated with human IL-18BP at 120 ng / mL for 30 min at room temperature. A 0.6 ng / mL recombinant human IL-18 stock solution (SinoBiological, 10119-HNCE) was then prepared and added to the antibody / IL-18BP complex after incubation. The resulting solution was immediately added to the cells to a final concentration of 15 μg / mL test antibody, 20 ng / mL human IL-18BP, and 0.1 ng / mL human IL-18. The cells were incubated at 37°C, 5% CO2 for 18-22 hours. 10 μL / well of cell supernatant was taken and mixed with 90 μL / well of complete Quanti-Blue solution (Invivogen) in a new 96-well plate and placed in an incubator at 37°C for 1-3 hours, noting a colorimetric change from purple to blue. The plate was then read at 620 nm and the data analyzed using GraphPad Prism.

[0459] Anti-IL-18BP mAb abolished the inhibition of IFNγ expression in KG-1 cells.

[0460] Human KG-1 cells (ATCC catalog number CCL-246) were plated with 150k cells / well. IL-18BP (ultimately 50 ng / mL, Sino Biologicals, catalog number 10357-H08H) was pre-blocked for 20 min at room temperature with antibody serial dilutions. IL-18 (ultimately 10 ng / mL, R&D Systems, catalog number 9124-IL / CF) was added to this mixture and incubated for another 20 min at room temperature. This mixture was added to the cells and incubated overnight at 37°C. Secreted IFNγ was measured in cell culture supernatants using the human IFN-γ DuoSet ELISA (catalog number DY285B) from R&D Systems on Nunc MaxiSorp flat-bottom plates (catalog number 44-2404-21). The manufacturer's protocol was followed and the supernatant was diluted 1:2 with reagent diluent. Absorbance was measured at 450 nm using a Spectramax iD5 plate reader. Data were analyzed using GraphPad Prism.

[0461] PBMC analysis of human IL-18 activity.

[0462] Human peripheral blood mononuclear cells (PBMCs) were obtained from the San Diego Blood Bank at a concentration of 2 × 10 5 Cells were seeded into 96-well flat-bottom plates (GenClone, catalog number 25-109) or at 1.7 × 10 5 10 cells / well were seeded into RPMI+GlutaMAX (Gibco catalog number 61870036) containing 100U penicillin, 100μg streptomycin (Gibco catalog number 10378016) and 10% FBS (RPMIc) in a round bottom plate (GenClone, catalog number 25-221). The test antibody was added to the wells at a volume of 50 μL at a final concentration of 4×. 50 μL of recombinant human IL-12 (R&D system, catalog number 219-IL-005) was then added at 4ng / mL, followed by 50 μL of recombinant human IL-18 (Sino Biological, catalog number 10119-HNCE) at 8ng / mL, reaching a final concentration of 1ng / mL and 2ng / mL, respectively. For analysis using cynomolgus macaque PBMCs (iQ Biosciences, catalog number IQB-MnPB102), 1.7×10 5 Cells were seeded at 100 cells / well. Recombinant cynomolgus macaque IL-12 (R&D Systems, catalog number 10215-CL) was added at a final concentration of 1 ng / mL, and recombinant rhesus macaque IL-18 (R&D Systems catalog number 2548-RM-025 / CF [note that the amino acid sequence of rhesus and cynomolgus macaque IL-18 is identical]) was added at a final concentration of 2 ng / mL. All dilutions were performed in RPMIc. The cells were incubated at 37°C, 5% CO2 for 48 hours. At 48 hours, a 50 μL aliquot of supernatant was removed from each well and analyzed for the presence of IFNγ using DuoSet ELISA kits (R&D Systems, catalog number DY285B for human IFNγ and catalog number DY961 for primate IFNγ) in Nunc MaxiSorp flat-bottom plates (Invitrogen, catalog number 44-2404-21) according to the manufacturer's instructions. CCL2 release was measured using a DuoSet ELISA kit (R&D Systems, catalog number DY279 for human CCL2) according to the manufacturer's instructions on NuncMax iSorp flat-bottom plates (Invitrogen, catalog number 44-2404-21). Absorbance was measured at 450 nm using a Spectramax iD5 plate reader and data were analyzed using GraphPad Prism software.

[0463] In the experiment where the reaction was carried out in the presence of pre-complexed hIL-18 / hIL-18BP, the design was as follows. 80 ng / mL of recombinant human or cynomolgus macaque IL-18 was incubated with recombinant human (SinoBiological, catalog number 10357-H08H) or cynomolgus macaque IL-18BP (produced in-house) at 400 ng / mL for 30 min at room temperature. Serial dilutions of mAb were added to a 96-well round-bottom plate at 50 μL / well at a 4× concentration. 50 μL / well of IL-18-IL-18BP complex was added to each well containing mAb and incubated at 37°C for 1 hour. After 1 hour, 50 μL / well of 4 ng / mL of recombinant human or cynomolgus macaque IL-12 and 2×10 6 50 μL of PBMCs containing 1×10 cells / mL were added to each well so that the final concentrations in each well were 20 ng / mL IL-18, 100 ng / mL IL-18BP, 1 ng / mL IL-12, and 1×10 5 PBMCs. All dilutions were performed in RPMI. Control wells contained either IL-12 + IL-18 alone or IL-12 + IL-18 + IL-18BP alone. Cells were incubated at 37°C, 5% CO₂ for 48 h, and supernatants were collected and analyzed for the presence of IFNγ and CCL2.

[0464] NK cell analysis of human IL-18 activity.

[0465] IL-18 activity was assessed using purified NK cells and pre-complexed hIL-18 / hIL-18BP. 10 ng / mL of recombinant human IL-18 (4 times the final concentration) was incubated with 50 ng / mL of recombinant human IL-18BP (4 times the final concentration) at room temperature for 30 min. NK cells were purified from fresh PBMC using the MojoSort Human NK Cell Isolation Kit (BioLegend, catalog number 480054) according to the manufacturer's instructions. NK purity was assessed by flow cytometry and was typically >90%. 1.7 × 10 5 NK cells were added to 50 μL of RPMI per well of a 96-well flat-bottom plate. Pre-complexed hIL-18 / hIL-18BP was added at 50 μL / well, followed by 50 μL of recombinant human IL-12 at 4 ng / mL (for a final concentration of 1 ng / mL) and incubated at 37°C for 30 minutes. Serial dilutions of the test antibody were added at 4-fold concentrations at 50 μL / well. The plates were incubated at 37°C, 5% CO₂ for 24 hours, and the supernatants were collected and analyzed for the presence of IFNγ by ELISA as described above.

[0466] result

[0467] Monoclonal antibody sequences were isolated from immunized mice.

[0468] DiversimAb TM and DivergimAb TM Groups of mice (Abveris) were immunized with human and cynomolgus macaque IL-18BP. The titers of human, cynomolgus macaque and mouse IL-18BP were measured by ELISA, and mice with high titers were selected to isolate antibodies to secretory B cells using the Berkeley Lights Beacon Optofluidic system (Mullen, 2021). The screen identified antibodies to secretory single cells that cross-reacted with human and cynomolgus macaque IL-18BP and some of which also bound to mouse IL-18BP. In addition, a novel screen was designed to identify antibodies to secretory cells that were unable to recognize forms of IL-18BP in which the active site had been blocked, referred to as "low IL-18". This screened out antibodies that could identify putative ligand-blocking antibodies.

[0469] Since IL-18 exhibits a high affinity for binding proteins (KD less than 1 nM) (Kim et al., PNAS 97(3):1190-1195, 2000; Kimura et al., Allergol Int. 57(4):367-76, 2008), mAbs with affinities in the nM affinity range (such as mAbs typically isolated from antigen-specific B cells) may not antagonize the IL-18 / IL-18BP interaction. Therefore, to evaluate the potential blocking ability of newly identified mAb candidates, a novel chimeric protein "low IL-18" was generated. Low IL-18 comprises human or mouse IL-18 tethered to its corresponding IL-18BP but separated by a flexible linker peptide (schematically shown in Figure 1B-1C The sequence of low IL-18 is provided in Table S1 below.

[0470]

[0471]

[0472] The rationale for this molecule is that the IL-18BP active site will be blocked because its tethered ligand will be unable to dissociate. Therefore, a mAb that recognizes the active binding site of BP will be sterically hindered from recognizing low IL-18, while most non-blocking antibodies will be able to bind equally to IL-18BP and low IL-18. A TEV protease cleavage site was also included in the design to provide the ability to separate hIL-18 from its binding protein.

[0473] Low IL-18 was designed based on a crystal structure (Protein Data Bank [PDB] structure 3F62) that included human IL-18 in complex with mousepox virus IL-18BP, which binds the C-terminus of IL-18 to the N-terminus of IL-18BP. Because orthopoxviruses, including mousepox, encode functional IL-18BP homologs that exhibit 17-34% amino acid identity with the mammalian orthologous IL-18BP (Calderara, 2001), it was believed that the 3F62 crystal structure would be instructive in the design of low IL-18 constructs. The resolved crystal structure indicated that the C-terminus of IL-18 was relatively close to the N-terminus of the binding protein, and therefore a fusion protein was possible. Thus, certain embodiments include a low IL-18 fusion protein comprising, in an N-terminal to C-terminal orientation, a signal peptide, IL-18, a first flexible linker, a protease cleavage site (optionally a TEV protease cleavage site), a flexible linker, and an IL-18BP; wherein the IL-18 portion of the fusion protein binds to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein. In specific embodiments, the low IL-18 fusion protein comprises, consists of, or consists essentially of an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% identical to a sequence from Table S1. Also included are nucleic acid molecules encoding the low IL-18 fusion protein.

[0474] The activity of the re-expressed mAbs was confirmed and analyzed for binding kinetics, blocking capacity, and biological activity.

[0475] Antibody sequences were derived from single B-cell screening on the Beacon instrument. To confirm activity, antibodies were recombinantly re-expressed using isolated murine variable region sequences fused to a human IgG1κ constant region. Antibodies were expressed in HEK293 cells and assayed or purified from cell culture supernatants for more detailed analysis.

[0476] Initial characterization used BLI to test binding affinity to human, cynomolgus macaque, and mouse IL-18BP. Additionally, mAbs were tested for binding to low IL-18 to identify mAbs most likely to be blocking antibodies. mAbs were also scored based on their expression and whether their sequence was considered problematic. Examples of problematic sequences include polytyrosine sequences in CDR3 or low levels of somatic hypermutation, which indicates that the antibody has not undergone significant in vivo maturation. The results are summarized in Table E1.

[0477]

[0478] The activity of the antibodies of interest was also tested in a cell-based reporter gene assay. This analysis uses an engineered cell line in which IL-18 signaling induces the secretion of alkaline phosphatase, which is easily measured. IL-18 induction of alkaline phosphatase is inhibited by adding IL-18BP, and this inhibition is alleviated by adding antibodies to IL-18BP, with the restriction that these antibodies can block IL-18BP / IL-18 binding. As shown in Table E2, several antibodies can block IL-18 / IL-18BP interaction and allow induction of IL-18-driven reactions. The antibodies that produced activity in this analysis were consistent with antibodies that could not bind low IL-18, thereby verifying the screening method for isolating the antibodies of interest.

[0479]

[0480] In addition, the biological activity of the antibody was tested in KG-1 cells. KG-1 is a human bone marrow-derived macrophage cell line that responds to IL-18 by producing IFNγ. The addition of IL-18BP blocks the ability of IL-18 to induce IFNγ expression, thereby inhibiting the reaction. Further addition of a neutralizing anti-IL-18BP antibody (which binds to IL-18BP) disrupts the interaction with IL-18, thereby releasing IL-18 to induce IFNγ. The antibody exhibits the ability to release IL-18 from IL-18BP inhibition in this analysis, as shown in Table E3.

[0481] Table E3

[0482]

[0483] mAb optimization

[0484] Two mAbs were prioritized for humanization and optimization: SA04a and SA44a (see Table E4).

[0485]

[0486] Humanization of SA04a.

[0487] The heavy chain CDRs of SA04a were transplanted into three different human variable regions: IGHV7-4-1*02, IGHV1-3*04, and IGHV3-23. The light chain CDRs were transplanted into hIGKV1-33*01 with framework backmutations P44V, F71Y, and Y87F. The heavy chain was transplanted into IGHV7-4-1*02 with framework backmutations G26V, V37L, L45F, and Y91F. The binding characteristics of the resulting humanized mAb (SA50a) are shown in Table E5 below.

[0488]

[0489] Humanization of SA44a.

[0490] DivergimAb TM The derived SA44a antibody's HC and LC CDRs were grafted into the human germline variable regions IgHV4-30-4*01 and IGKV3-11*01, respectively. This grafting procedure significantly reduced binding affinity. Therefore, to optimize affinity, two back mutations (G27Y in FW1 and V71R in FW3) were reintroduced into the HC, and one back mutation (L46P in FW2) was reintroduced into the LC. As shown in Table E6 below, the resulting humanized mAb (SA301a) restored full activity and superior expression relative to its original murine parent (SA44a).

[0491]

[0492] Maturation of SA301a.

[0493] To affinity mature mAb SA301a, we generated a library of mAb variants centered around the HC and LC CDRs. To achieve this, CDR amino acids were sequentially replaced with up to 17 amino acid substitutions—cysteine ​​and tryptophan were excluded from the library to avoid introducing undesirable potential sequence biases, and the parent amino acid was not already present in the positions included in the screen. Following generation, the variants were first screened by BLI at a single concentration of human and cynomolgus macaque IL-18BP to determine if any had improved on- or off-rates compared to the parent antibody SA301a. The screen identified multiple single-point variants that did indeed improve apparent binding kinetics. Individual mutations may be able to recombine to further improve binding affinity for the target antigen, but this was not the case in all cases.

[0494] The initial screen identified a subset of amino acid substitutions that specifically improved binding to both human and cynomolgus monkey IL-18BP. Therefore, these changes were recombined in various combinations for further analysis.

[0495] Table E7 shows variants that resulted in improved binding to human and cynomolgus macaque IL-18BP. The parenthetical (N) value indicates how many data points from independently run experiments were included in the mean; ± indicates SEM; not tested, NT; ND, not determined. In cases where a mAb was tested only twice, two values ​​are provided. Cynomolgus macaque / human represents the difference in ratio between the two species.

[0496]

[0497]

[0498]

[0499] Further characterization of high-affinity mAbs to human and cynomolgus macaque IL-18BP.

[0500] MAbs with high affinity for human and cynomolgus macaque IL-18BP were further characterized to identify MAbs suitable for further development. This included evaluating the antibody binding characteristics (e.g., blocking IL-18BP-mediated IL-18 signaling and neutralizing anti-human IL-18BP; human IgG1 / κ; cross-reactivity with cynomolgus macaque IL-18BP; high affinity KD for human IL-18BP, lower than the affinity of IL-18 for IL-18BP, and no detectable nonspecific binding or binding to homologous proteins), functional properties (e.g., blocking IL-18 binding to IL-18BP; neutralizing IL-18BP, thereby antagonizing its inhibitory effect on IL-18-mediated IFNγ induction), and developability.

[0501] Functional activity.

[0502] The ability of the relevant mAbs to inhibit IL-18 neutralization by IL-18BP was evaluated in a variety of different bioassays. These bioassays included activity against IL-18 activity in two different types of PBMCs. The experimental details of these analyses are described in the Materials and Methods section above.

[0503] IL-18 binds to IL-12 to induce multiple cell types (including NK cells and T cells) in PBMC to produce IFNγ. IL-18BP is endogenously produced by PBMC and is upregulated by IFNγ, thereby providing a negative feedback loop that inhibits IFNγ reactions. Adding neutralizing anti-IL-18BP antibodies will bind to endogenous IL-18BP and prevent the negative feedback loop, thereby causing the induction of IFNγ production. A group of mAbs were tested in vitro in healthy donor PBMCs to compete with IL-18 for binding to IL-18BP and allow IL-18-mediated IFNγ induction. The concentration of IL-18 used in the analysis was only sufficient to provide a low-level IFNγ response in PBMC, which may be due to inhibition by endogenous IL-18BP.

[0504] As shown in Table E8 and Figure 2A-2G As shown in , addition of mAb removed this inhibition, leading to a dose-dependent increase in IFNγ production. The potency of the antibody correlated broadly, but not in all cases, with its binding affinity for IL-18BP.

[0505] In addition, a second assay format was performed using human PBMCs. In this format, IL18 was pre-complexed with IL-18BP and then added to the PBMCs. This assay tested the ability of the antibody to release IL-18 from a pre-existing complex with IL-18BP. Figure 3A-3GActivity was also observed in this assay, with the potency of IFNγ induction again correlating in most cases with the affinity of the highest affinity mAb. EC50 values ​​are provided in Table E8 below.

[0506] Table E8 Activity of mAbs in PBMC assay compared to affinity (KD)

[0507]

[0508] Anti-human IL-18BP antibodies can also induce IL-18 to produce additional cytokines from PBMCs, such as CCL2. The antibodies were tested in PBMC assay 1 (endogenous IL-18BP assay described above) and CCL2 release was determined by ELISA. Addition of anti-IL-18BP mAb resulted in an increase in CCL2 production by PBMCs ( Figure 4 ).

[0509] The activity of anti-IL-18BP mAb was also tested using a cynomolgus macaque PBMC assay. PBMC isolated from cynomolgus macaque blood were used to perform the two assays described above for human PBMC. The antibody exhibited activity in both assays, as described in Table 1. Figure 5A-5B As shown in .

[0510] Activity was also tested in an NK cell specific assay. In this assay, purified human NK cells were incubated with pre-complexed IL-18:IL-18BP in the presence and absence of test antibodies. The test antibodies were able to disrupt the IL-18:IL-18BP complex and induce NK cell secretion of IFNγ, such as Figure 6 As shown in .

[0511] Library screening data

[0512] Variants of the parental SA301a and SA302a antibodies were tested for their binding characteristics.

[0513] Heavy chain library screening.

[0514] Octet-derived kinetic values ​​measured at >50 nM are listed as "Inactive". Consider providing the product for humans and cynomolgus monkeys (when tested). The position of 2.0-fold improvement was taken for further analysis. The values ​​presented for each species are apparent KDs, as they are calculated based on a single binding concentration collected in a semi-high-throughput format. Samples were compared to the parental background from which they were generated (font is normal in the case of SA301a parent and italicized in the case of SA302a parent with HC-C34A).

[0515]

[0516]

[0517]

[0518]

[0519]

[0520]

[0521]

[0522]

[0523] *This particular well was contaminated and the purified parent was used in its place.

[0524]

[0525]

[0526]

[0527]

[0528]

[0529] Light chain library screening.

[0530] Octet-derived kinetic values ​​measured at >50 nM are listed as "inactive." The values ​​presented for each species are apparent KD values, as they were calculated from a single binding concentration collected in a semi-high-throughput format. Samples were compared to the parental background from which they were derived, i.e., font is normal when the parent is SA301a and italic when the parent is SA326a (HC-C34A x LC-Q90L).

[0531]

[0532]

[0533]

[0534]

[0535]

[0536]

[0537]

[0538]

[0539] Epitope mapping

[0540] The epitope of mAb SA338a on IL-18BP was determined by a cross-linking / high-resolution mass spectrometry method developed by CovalX AG (Pimenova et al., 2008, J. Mass Spectrometry 43:185). Briefly, human IL-18BP was conjugated to SA338a and cross-linked with a heterobifunctional linker. The resulting complex was digested with five different proteases (trypsin, chymotrypsin, ASP-N, elastase, and thermolysin), and the resulting peptides, whether cross-linked or not, were analyzed by high-resolution mass spectrometry.

[0541] The results show that SA338a recognizes conformational epitopes, including residues in the IL-18BP indicated below. In addition, the residues on the IL-18BP that interact with IL-18 were similarly located using the same technique. The following results show that the four residues on IL-18BP that are related to the recognition of IL-18 are also the residues recognized by SA338a, supporting the evidence that this is a functional blocking antibody (S75, H79, T116 and S119). SA338a interacts with the following residues on IL-18BP: T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, S119. The residues on IL-18BP that interact with IL-18 are identified as: R61, Y69, S75, H79, T116, S119 and R131. The sequence of mature IL-18BP is shown below, with residues interacting with SA338a and IL-18 highlighted as indicated.

[0542]

[0543] The residues found to interact with IL-18 are highlighted in bold and underlined. The residues forming the epitope of SA338 are highlighted in bold and italic. Residues S75, H79, T116, and S119 are recognized by both IL-18 and SA338a and are shown in bold, italic, and underlined.

Claims

1. An antibody specific for interleukin-18 binding protein (IL-18BP), wherein the antibody interferes with the binding of IL-18 to IL-18BP.

2. The antibody of claim 1, wherein the antibody binds to a conformational epitope of IL-18BP.

3. The antibody of claim 2, wherein the antibody binds to two or more of amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO:

372.

4. The antibody of claim 2, wherein the antibody binds to amino acid residues T51, S53, S75, H79, R83, S88, S90, T110, H114, S115, T116, and S119 of SEQ ID NO:

372.

5. The antibody of claim 1, wherein the antibody binds to a linear epitope of IL-18BP.

6. The antibody of any one of claims 1 to 5, wherein the antibody binds to the binding interface between IL-18 and the mature form of IL-18BP.

7. The antibody of claim 6, wherein the antibody binds to amino acid residues S75, H79, T116, S119 of SEQ ID NO:

372.

8. The antibody of any one of claims 1 to 7, wherein the antibody binds to human IL-18BP and cynomolgus monkey IL-18BP, but does not bind to mouse IL-18BP.

9. The antibody of any one of claims 1 to 7, wherein the antibody binds to human IL-18BP, cynomolgus monkey IL-18BP, and mouse IL-18BP.

10. The antibody according to any one of claims 1 to 9, wherein the antibody binds to IL-18 with a binding affinity (K) greater than that between IL-18 and IL-18BP. D 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM), optionally binds to IL-18BP with a binding affinity of about 1 pM to about 650 pM, or about or less than about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, or 300, 400, 500, 600, or 650 pM. The antibody according to any one of claims 1 to 10, wherein the antibody is an IL-18BP antagonist that antagonizes the binding activity between IL-18BP and IL-18.

12. The antibody of claim 11, wherein the antibody blocks the inhibitory activity of IL-18BP on IL-18 and thereby increases IL-18-mediated signaling, including the induction of IFN-γ, CXCL10 and / or TNFα.

13. The antibody according to any one of claims 1 to 12, comprising an IgA (including subclasses IgA1 and IgA2), IgD, IgE, IgG (including subclasses IgG1, IgG2, IgG3 and IgG4) or IgM Fc domain, optionally a human Fc domain or a hybrid and / or variant thereof.

14. The antibody according to claim 13, comprising an IgG Fc domain with high effector function in humans, optionally an IgG1 or IgG3 Fc domain.

15. The antibody of claim 13, comprising an IgG Fc domain with low effector function in humans, optionally an IgG2 or IgG4 Fc domain.

16. The antibody of any one of claims 1 to 15, wherein the antibody is a monoclonal antibody.

17. The antibody according to any one of claims 1 to 16, wherein the antibody is a humanized antibody.

18. The antibody of any one of claims 1 to 17, wherein the antibody is a scFv.

19. The antibody of any one of claims 1 to 18, wherein the antibody comprises: a. Heavy chain variable region (V H ), which comprises a complementarity determining region V selected from Table A1 H CDR1, V H CDR2 and V H CDR3 sequences and variants thereof that specifically bind to IL-18BP; and b. Light chain variable region (V L ), which comprises a complementarity determining region V selected from Table A1 L CDR1, V L CDR2 and V L CDR3 sequences and their variants that specifically bind to IL-18BP.

20. The antibody of claim 19, wherein: a. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 1-3, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 4-6, respectively; b. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 7-9, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 10-12, respectively; c. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 13-15, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 16-18, respectively; d. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 19-21, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 22-24, respectively; e. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 25-27, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 28-30, respectively; f. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 31-33, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 34-36, respectively; g. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 37-39, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 40-42, respectively; h. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 43-45, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 46-48, respectively; i. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 49-51, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 52-54, respectively; j. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 55-57, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 58-60, respectively; k. V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 61-63, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 64-66, respectively; 1. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 67-69, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 70-72, respectively; m.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 73-75, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 76-78, respectively; n.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 79-81, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 82-84, respectively; o. Said V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 85-87, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 88-90, respectively; p.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 91-93, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 94-96, respectively; q. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 97-99, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 100-102, respectively; r. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 103-105, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 106-108, respectively; s.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 109-111, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 112-114, respectively; t.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 115-117, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 118-120, respectively; u.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 121-123, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 124-126, respectively; v.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 127-129, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 130-132, respectively; w.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 133-135, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 136-138, respectively; x.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 139-141, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 142-144, respectively; y. Said V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 145-147, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 148-150, respectively; z.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 151-153, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 154-156, respectively; aa.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 157-159, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 160-162, respectively; bb.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 163-165, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 166-168, respectively; cc.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 169-171, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 172-174, respectively; dd.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 175-177, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 178-180, respectively; ee.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 181-183, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 184-186, respectively; ff. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 187-189, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 190-192, respectively; gg.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 193-195, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 196-198, respectively; hh.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 199-201, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 202-204, respectively; ii. V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 205-207, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 208-210, respectively; jj.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 211-213, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 214-216, respectively; kk.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 217-219, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 220-222, respectively; ll. The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 223-225, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 226-228, respectively; mm.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 229-231, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 232-234, respectively; nn.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 235-237, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 238-240, respectively; oo.The V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 241-243, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 244-246, respectively; pp.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 247-249, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 250-252, respectively; qq.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 253-255, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 256-258, respectively; or rr.V H CDR1, V H CDR2 and V H The CDR3 sequences comprise SEQ ID NOs: 259-261, respectively, and the V L CDR1, V L CDR2 and V L The CDR3 sequences comprise SEQ ID NOs: 262-264, respectively.

21. The antibody according to any one of claims 1 to 20, wherein the V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein said V H There are 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in the framework regions.

22. The antibody according to any one of claims 1 to 21, wherein the V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to a sequence selected from Table A2, optionally wherein said V L There are 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations in the framework regions.

23. The antibody according to any one of claims 1 to 22, wherein: a. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 265, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 266; b. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 267, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 268; c. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 269, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 270; d. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 271, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 272; e. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 273, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 274; f. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 275, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 276; g. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 277, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 278; h. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 279, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 280; i. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 281, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 282; j. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 283, and wherein said V L comprising a sequence at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 284; k. V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 285, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 286; 1. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 287, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 288; m.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 289, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 290; n.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 291, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 292; o. Said V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 293, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 294; p.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 295, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 296; q. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 297, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 298; r. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 299, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 300; s.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 301, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 302; t.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 303, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 304; u.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 305, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 306; v.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 307, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 308; w.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 309, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 310; x.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 311, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 312; y. Said V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 313, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 314; z.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 315, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 316; aa.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 317, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 318; bb.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 319, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 320; cc.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 321, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 322; dd.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 323, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 324; ee.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 325, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 326; ff. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 327, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 328; gg.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 329, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 330; hh.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 331, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 332; ii. V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 333, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 334; jj.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 335, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 336; kk.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 337, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 338; ll. The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 339, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 340; mm.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 341, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 342; nn.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 343, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 344; oo.The V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 345, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 346; pp.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 347, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 348; qq.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 349, and wherein said V L comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 350; or rr.V H comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 351, and wherein said V L Comprising a sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO:

352.

24. An isolated polynucleotide encoding the isolated anti-IL-18BP antibody of any one of claims 1 to 23; an expression vector comprising the isolated polynucleotide; or an isolated host cell comprising the vector.

25. A pharmaceutical composition comprising the anti-IL-18BP antibody according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

26. The pharmaceutical composition of claim 25, wherein the composition is a sterile injectable solution optionally suitable for intravenous, intramuscular, subcutaneous or intraperitoneal administration.

27. A method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of any one of claims 25 to 26.

28. The method of claim 27, wherein the disease or condition is cancer or a tumor or a proliferative disease or disorder, optionally selected from the group consisting of a lymphoproliferative disorder, a myeloproliferative disorder, a proliferative enteritis, a proliferative diabetic retinopathy, and a proliferative nephropathy.

29. The method of claim 28, wherein the cancer or tumor expresses or overexpresses IL-18BP and / or IL-18, or wherein the proliferative disease or disorder is associated with increased expression of IL-18BP and / or IL-18.

30. The method of claim 28 or 29, wherein the cancer is one or more selected from the group consisting of bone cancer, prostate cancer, melanoma (e.g., metastatic melanoma), pancreatic cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), mesothelioma, leukemia (e.g., lymphocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, relapsed acute myeloid leukemia, hairy cell leukemia, acute lymphoblastic leukemia), lymphoma (e.g., non-Hodgkin's lymphoma), lymphoma), Hodgkin's lymphoma), liver cancer (hepatocellular carcinoma), sarcoma, B-cell malignancies, breast cancer, ovarian cancer, colorectal cancer, glioma, glioblastoma multiforme, meningioma, pituitary adenoma, vestibular schwannoma, primary CNS lymphoma, primitive neuroectodermal tumor (medulloblastoma), kidney cancer (e.g., renal cell carcinoma), bladder cancer, urothelial cancer, uterine cancer, esophageal cancer, brain cancer, head and neck cancer, cervical cancer, testicular cancer, thyroid cancer, and stomach cancer.

31. The method according to any one of claims 27 to 30, comprising administering a pharmaceutical composition according to claim 25 or 26 in combination with IL-18.

32. The method of any one of claims 28 to 31, comprising administering a combination of a pharmaceutical composition according to claim 25 or 26 and an immune checkpoint regulator selected from antagonists of inhibitory immune checkpoint molecules and agonists of stimulatory immune checkpoint molecules.

33. The method of claim 32, wherein the immune checkpoint regulator is a polypeptide, optionally an antibody or a ligand or a small molecule.

34. The method of claim 32 or 33, wherein the inhibitory immune checkpoint molecule is one or more selected from the group consisting of programmed death-ligand 1 (PD-L1), programmed death 1 (PD-1), programmed death-ligand 2 (PD-L2), cytotoxic T lymphocyte-associated protein 4 (CTLA-4), indoleamine 2,3-dioxygenase (IDO), tryptophan 2,3-dioxygenase (TDO), T cell immunoglobulin domain and mucin domain 3 (TIM-3), lymphocyte activation gene-3 (LAG-3), V-domain Ig inhibitory factor of T cell activation (VISTA), B and T lymphocyte attenuator (BTLA), CD160, herpes virus entry mediator (HVEM), and T cell immunoreceptor with Ig and ITIM domains (TIGIT).

35. The method of claim 34, wherein: a. the antagonist is a PD-L1 and / or PD-L2 antagonist, optionally selected from one or more of the following: an antibody or small molecule that specifically binds to PD-L1 and / or PD-L2, atezolizumab (MPDL3280A), avelumab (MSB0010718C), and durvalumab (MEDI4736), optionally wherein the cancer is selected from one or more of the following: colorectal cancer, melanoma, breast cancer, non-small cell lung cancer, bladder cancer, and renal cell carcinoma; b. the antagonist is a PD-1 antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to PD-1, nivolumab, pembrolizumab, MK-3475, AMP-224, AMP-514PDR001, and pidilizumab, optionally wherein the PD-1 antagonist is nivolumab, and the cancer is optionally selected from one or more of the following: Hodgkin's lymphoma, melanoma, non-small cell lung cancer, hepatocellular carcinoma, renal cell carcinoma, and ovarian cancer; c. the PD-1 antagonist is pembrolizumab, and the cancer is optionally selected from one or more of the following: melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, and urothelial carcinoma; d. the antagonist is a CTLA-4 antagonist, optionally selected from one or more of the following: an antibody or small molecule that specifically binds to CTLA-4, ipilimumab, tremelimumab, optionally wherein the cancer is selected from one or more of the following: melanoma, prostate cancer, lung cancer, and bladder cancer; e. the antagonist is an IDO antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to IDO, indoximod (NLG-8189), 1-methyl-tryptophan (1MT), a β-carboline (norharmane; 9H-pyrido[3,4-b]indole), rosmarinic acid, and epacadostat, and wherein the cancer is optionally selected from one or more of the following: metastatic breast cancer and brain cancer, optionally glioblastoma multiforme, glioma, gliosarcoma, or a malignant brain tumor; f. The antagonist is a TDO antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to TDO, 680C91, and LM10; g. the antagonist is a TIM-3 antagonist optionally selected from one or more antibodies or small molecules that specifically bind to TIM-3; h. the antagonist is a LAG-3 antagonist optionally selected from one or more of the following: an antibody or small molecule that specifically binds to LAG-3, and BMS-986016; i. the antagonist is a VISTA antagonist optionally selected from one or more antibodies or small molecules that specifically bind to VISTA; j. the antagonist is a BTLA, CD160 and / or HVEM antagonist optionally selected from one or more antibodies or small molecules that specifically bind to BTLA, CD160 and / or HVEM; or k. The antagonist is a TIGIT antagonist, optionally selected from one or more antibodies or small molecules that specifically bind to TIGIT.

36. The method of claim 32 or 33, wherein the stimulatory immune checkpoint molecule is one or more selected from the group consisting of OX40, CD40, glucocorticoid-induced TNFR family-related genes (GITR), CD137 (4-1BB), CD27, CD28, CD226, and herpes virus entry mediator (HVEM).

37. The method of claim 36, wherein: a. The agonist is an OX40 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to OX40, OX86, Fc-OX40L and GSK3174998; b. the agonist is a CD40 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD40, CP-870,893, dacetuzumab, Chi Lob 7 / 4, ADC-1013, and rhCD40L, and wherein the cancer is optionally selected from one or more of the following: melanoma, pancreatic cancer, mesothelioma, and a blood cancer, optionally a lymphoma, such as non-Hodgkin's lymphoma; c. the agonist is a GITR agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to GITR, INCAGN01876, DTA-1 and MEDI1873; d. the agonist is a CD137 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD137, utomilumab, and a 4-1BB ligand; e. the agonist is a CD27 agonist optionally selected from one or more of the following: an antibody or small molecule or ligand that specifically binds to CD27, varlilumab, and CDX-1127 (1F5); f. the agonist is a CD28 agonist optionally selected from one or more of the following: an antibody or a small molecule or a ligand that specifically binds to CD28, and TAB08; and / or g. The agonist is an HVEM agonist optionally selected from one or more antibodies or small molecules or ligands that specifically bind to HVEM.

38. The method of any one of claims 28 to 37, comprising administering a pharmaceutical composition according to claim 25 or 26 in combination with at least one chemotherapeutic agent.

39. The method of claim 38, wherein the at least one chemotherapeutic agent is one or more selected from the group consisting of an alkylating agent, an antimetabolite, a cytotoxic antibiotic, a topoisomerase inhibitor (Type 1 or Type II), and an anti-microtubule agent.

40. The method of claim 39, wherein: a. The alkylating agent is one or more selected from the group consisting of nitrogen mustard (optionally mechlorethamine, cyclophosphamide, mustine, melphalan, chlorambucil, ifosfamide, and busulfan), nitrosourea (optionally N-nitroso-N-methylurea (MNU), carmustine (BCNU), lomustine (CCNU), semustine (MeCCNU), fotemustine (fotemustine), and methylprednisolone. e) and streptozotocin), tetrazines (optionally dacarbazine, mitozolomide and temozolomide), aziridines (optionally thiotepa, mytomycin and diaziquone (AZQ)), cisplatin and its derivatives (optionally carboplatin and oxaliplatin) and atypical alkylating agents (optionally procarbazine and hexamethonium); b. the antimetabolite is one or more selected from the group consisting of antifolates (optionally methotrexate and pemetrexed), fluoropyrimidines (optionally 5-fluorouracil and capecitabine), deoxynucleoside analogs (optionally ancitabine, enocitabine, cytarabine, gemcitabine, decitabine, azacitidine, fludarabine, nelarabine, cladribine, clofarabine, fludarabine, and pentostatin), and thiopurines (optionally thioguanine and mercaptopurine); c. The cytotoxic antibiotic is one or more selected from the group consisting of anthracyclines (optionally, doxorubicin, daunorubicin, epirubicin, idarubicin, pirarubicin, aclarubicin and mitoxantrone), bleomycin, mitomycin C, mitoxantrone and actinomycin; d. the topoisomerase inhibitor is one or more selected from the group consisting of camptothecin, irinotecan, topotecan, etoposide, raspberry, mitoxantrone, teniposide, novobiocin, merbarone and aclarubicin; and / or e. The anti-microtubule agent is one or more selected from the group consisting of taxanes (optionally, paclitaxel and docetaxel) and vinca alkaloids (optionally vinblastine, vincristine, vindesine, vinorelbine).

41. The method of claim 27, wherein the disease or condition is myelodysplastic syndrome (MDS).

42. The method of claim 27, wherein the disease or condition is an infectious disease.

43. The method of claim 42, wherein the infectious disease is selected from viral, bacterial, fungal (optionally yeast) and protozoal infections.

44. The method of any one of claims 41 to 43, comprising administering a pharmaceutical composition according to claim 25 or 26 in combination with IL-18.

45. A method for screening anti-IL-18BP antibodies for their ability to block or inhibit the binding between IL-18 and IL-18BP, comprising a. determining the binding affinity of the antibody to (i) IL-18BP alone and (ii) a low IL-18 fusion protein, wherein the low IL-18 fusion protein comprises IL-18 fused to IL-18BP via a flexible linker (and optionally a protease cleavage site therebetween), wherein the IL-18 portion of the fusion protein binds to the IL-18BP portion of the fusion protein and sterically blocks the IL-18 binding site of the IL-18BP portion of the fusion protein; b. comparing the binding affinity of (i) to the binding affinity of (ii); and c. If the binding affinity of (i) is significantly stronger than the binding affinity of (ii), then the antibody is identified or selected as being capable of blocking or inhibiting the binding between IL-18 and IL-18BP.

46. ​​The method of claim 45, wherein the IL-18 and the IL-18BP are mouse IL-18 and mouse IL-18BP.

47. The method of claim 45, wherein the IL-18 and the IL-18BP are human IL-18 and human IL-18BP.

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