Bispecific antibody

Abstract

The present invention addresses the problem of providing a novel drug for preventing cancer, suppressing the progression of a symptom of cancer, suppressing the recurrence of cancer, or treating cancer.　As the results of intensive studies, the present inventors focused attention on Allergin-1 that is a target molecule involved in the regulation of cancer immunity, and found a bispecific antibody capable of binding specifically respectively to two different epitopes on the S1 and S2 domains, said domains being the extracellular domains of Allergin-1, as a substance that can solve this problem. The present inventors confirmed that this antibody is useful for preventing cancer, suppressing the progression of a symptom of cancer, suppressing the recurrence of cancer, or treating cancer.

Description

bispecific antibody

[0001] The present invention relates to a bispecific antibody or an antibody fragment thereof against Allergin-1, a pharmaceutical composition comprising the bispecific antibody as an active ingredient, and medical uses thereof.

[0002] Unlike conventional treatments such as surgery, radiation therapy, and chemotherapy with anticancer drugs or molecular-targeted drugs, cancer immunotherapy inhibits or treats cancer progression by acting on the cancer patient's own inherent immune surveillance system and strengthening the patient's anti-cancer immune system. Recent research into cancer immunology has revealed that cancer progression is related to an immunosuppressive environment centered on the tumor itself, and that cancer itself utilizes a system to evade the immune surveillance system. Molecules used in this evasion system include so-called immune checkpoint molecules such as CTLA-4, PD-1, and their ligand PD-L1, and inhibitors of these molecules have already achieved remarkable results in clinical trials.

[0003] However, it is true that there are still cancer patients for whom these immune checkpoint inhibitors do not provide sufficient therapeutic benefit, and there is an urgent need to develop new treatments for these cancer patients and to elucidate the functions of the target molecules that are essential for establishing such treatments.

[0004] Allergin-1 according to the present invention is a membrane receptor with an ITIM domain in its intracellular region and an immunoglobulin-like structure on the extracellular surface, and is a molecule that is highly expressed in mast cells. Analysis using mice lacking this molecule has shown that it functions as a molecule that suppresses allergic reactions, such as anaphylaxis by inhibiting IgE receptor signaling-mediated degranulation and asthma responses induced by mite antigens (Non-Patent Documents 1, 2, and 3).

[0005] On the other hand, Allergin-1 is also involved in cancer immunity, and its inhibition has been reported as a cancer treatment (Patent Document 1).

[0006] International Publication No. 2017 / 038997 Brochure

[0007] Nature Immunology, 2010, Vol. 11, No. 7, pp. 601-608; PLOS ONE, 2013, Vol. 8, No. 10, e76160; Allergy and Immunology, 2011, Vol. 18, No. 4, pp. 506-514

[0008] An object of the present invention is to provide a novel drug that has a cancer immunity enhancing effect and is effective in cancer treatment.

[0009] As a result of extensive research, the inventors of the present invention found that a bispecific antibody capable of specifically binding to two different epitopes in the extracellular domain of Allergin-1 (hereinafter sometimes abbreviated as "bispecific antibody against Allergin-1" or "anti-Allergin-1 bispecific antibody") can solve the above-mentioned problems, and thus completed the present invention.

[0010] That is, the present invention mainly provides the following embodiments: <1> A bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1 (hereinafter, these may be collectively referred to as "anti-Allergin-1 bispecific antibody, etc."); <2> A pharmaceutical composition comprising, as an active ingredient, a bispecific antibody against Allergin-1; <3> An agent for suppressing the progression of symptoms or recurrence of and / or treating cancer, comprising, as an active ingredient, a bispecific antibody against Allergin-1; <4> An antibody or antibody fragment thereof that specifically binds to the S1 domain of Allergin-1, or an antibody or antibody fragment thereof that specifically binds to the S2 domain of Allergin-1; and <5> Polynucleotides encoding the heavy chain and light chain of the bispecific antibody according to <1> above.

[0011] The present invention provides anti-Allergin-1 bispecific antibodies as novel agents for suppressing the progression of cancer symptoms, suppressing recurrence, and / or treating cancer.

[0012] The figure shows the correspondence between each amino acid sequence of the heavy chain variable region (hereinafter sometimes abbreviated as "VH") and light chain variable region (hereinafter sometimes abbreviated as "VL") that constitute the S1-A binding site and SEQ ID NO. As will be apparent to those skilled in the art, the alphabets in the figure represent single-letter codes corresponding to each amino acid. The figure shows the correspondence between each amino acid sequence of the VH and VL that constitute the S1-B binding site and SEQ ID NO. The figure shows the correspondence between each amino acid sequence of the multiple CDRs contained in the VH and VL that constitute the S1-A binding site and SEQ ID NO. The figure shows the correspondence between each amino acid sequence of the multiple CDRs contained in the VH and VL that constitute the S1-B binding site and SEQ ID NO. The figure shows the correspondence between each amino acid sequence of the VH and VL that constitute the S2-C binding site and S2-D binding site and SEQ ID NO. The figure shows the correspondence between each amino acid sequence of the multiple CDRs contained in the VH and VL that constitute the S2-C binding site and S2-D binding site and SEQ ID NO. The figure shows the Morrison form of the anti-Allergin-1 bispecific antibody of the present invention. 1 shows the correspondence between each amino acid sequence of the heavy chain constant region and light chain constant region constituting the anti-Allergin-1 bispecific antibody of the present invention and SEQ ID NO. 1 shows the correspondence between each amino acid sequence of the heavy chain ... light chain constituting the anti-Allergin-1 bispecific antibody of the present invention and SEQ ID NO. 1 shows the correspondence between each amino acid sequence of the light chain constituting the anti-Allergin-1 bispecific antibody of the present invention and SEQ ID NO. 1 shows the amino acid sequence of human Allergin-1L (GenBank accession number: NP_001078892.1). 1 shows the results of a competitive assay demonstrating the degree of shielding by the binding of the anti-Allergin-1 bispecific antibody of the present invention to Allergin-1.The multiple numerical values ​​in the table represent the binding rate of the anti-Allergin-1 antibody to Allergin-1L when treated with the anti-Allergin-1 bispecific antibody of the present invention, relative to the binding of the anti-Allergin-1 antibody to Allergin-1L when not treated with the same anti-Allergin-1 bispecific antibody. The numbers in the vertical column labeled "Rabbit IgG ID No." represent the clones of the anti-Allergin-1 antibodies used in each evaluation, with "anti-Allergin-1S1" indicating that the anti-Allergin-1 antibody is specific to the S1 domain and "anti-Allergin-1S2" indicating that the anti-Allergin-1 antibody is specific to the S2 domain. The numbers in the horizontal column to the right represent each clone of the anti-Allergin-1 bispecific antibody evaluated. The results of the evaluation of the binding ability of the anti-Allergin-1 bispecific antibody of the present invention to the S1 and S2 domains of Allergin-1 are shown. The vertical axis represents "referenced OD." ４５０－６９０ ｎｍ ​" represents the amount of anti-Allergin-1 bispecific antibody bound to Allergin-1L, and the horizontal axis represents the concentration of rabbit anti-Allergin-1 antibody added. In the figure, "Rabbit IgG #50" and "Rabbit IgG #85" represent the rabbit anti-Allergin-1S2 IgG antibody clone #50 and the rabbit anti-Allergin-1S1 IgG antibody clone #85, respectively. This figure shows the results of Biacore analysis of the inhibitory effect of the anti-Allergin-1 bispecific antibody of the present invention on the binding of the TLR9 ligand ODN M362 to Allergin-1. The vertical axis, "Response (RU)," represents the amount of ODN M362 bound to Allergin-1L. This figure shows the anti-tumor effects of the anti-Allergin-1 bispecific antibody of the present invention in combination with the anti-mouse PD-1 antibody 4H2 in a cancer-bearing model (mouse colon cancer cell line MC38) in human Allergin-1L knock-in C57BL / 6 mice. The vertical axis represents tumor volume, and the horizontal axis represents the number of days since tumor cell line transplantation. Figure 1 shows the results of the anti-tumor effect of combined use of the anti-Allergin-1 bispecific antibody of the present invention and the anti-mouse PD-1 antibody 4H2 in a cancer-bearing model (mouse lung cancer cell line 3LL) in human Allergin-1L knock-in C57BL / 6 mice. The vertical axis represents tumor volume, and the horizontal axis represents the number of days since tumor cell line transplantation. Figure 1 shows the results of the anti-tumor effect of single administration of the anti-Allergin-1 bispecific antibody of the present invention in a cancer-bearing model (mouse colon cancer cell line MC38 with forced expression of human PD-L1) in human Allergin-1L knock-in C57BL / 6 mice. The vertical axis represents tumor volume, and the horizontal axis represents the number of days since tumor cell line transplantation. Figure 1 shows the results of evaluation of the effect of the anti-Allergin-1 bispecific antibody of the present invention on cytokine release from human peripheral blood mononuclear cells.

[0013] Allergin-1, also known as MILR1, is composed of at least three splicing variants in humans: Allergin-1L, Allergin-1S1, and Allergin-1S2 (see Nature Immunology (2010), Vol. 11, No. 7, pp. 601-608). These variants are membrane-associated proteins with the amino acid sequences shown in GenBank accession numbers NP_001078892.1 (SEQ ID NO: 115), BAJ08252.1, and BAJ08253.1, respectively. Its mouse homolog, MCA32, has the amino acid sequence shown in GenBank accession number BAJ08254.1. Allergin-1L contains the S1 and S2 domains in its extracellular domain, while Allergin-1S1 and Allergin-1S2 contain only the S1 and S2 domains, respectively. The S1 domain of Allergin-1L is located in the region of 37 to 121 bases from its N-terminus, and the S2 domain is located in the region of 128 to 200 bases from its N-terminus. Similarly, the S1 domain of Allergin-1S1 is located in the region of 37 to 121 bases from its N-terminus, and the S2 domain of Allergin-1S2 is located in the region of 38 to 110 bases from its N-terminus. When the term "Allergin-1" is used herein, it may be used to include all of its splicing variants or isoforms, unless otherwise specified. In the present invention, the preferred Allergin-1 is human Allergin-1.

[0014] As used herein, "isolated" means extracted from a host cell and identified, separated, and / or purified from contaminants, including multiple or numerous components, to provide a substantially single, pure component.

[0015] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous antibody population that binds to the same specific antigen. As used herein, the term "bispecific antibody" refers to an antibody that has binding specificities for two different antigen molecules or epitopes in a single molecule. As used herein, the terms "antigenic molecule" or "antigen" refer to a chemical substance, peptide, or protein that induces an immune response in the body and is recognized by antibodies produced as part of the immune response, and the antigen of the present invention corresponds to the extracellular domain of Allergin-1 or a portion thereof. As used herein, the term "epitope" refers to a portion of an antigen that is recognized as a binding site when an antibody binds to the antigen, and the epitope of the present invention is a portion of the extracellular domain of Allergin-1 to which each antigen-binding site in the bispecific antibody or antibody fragment thereof of the present invention specifically binds.

[0016] As used herein, the term "antigen-binding site" refers to the smallest unit of an antibody that allows the antibody to bind to its antigen or epitope, and is usually formed by the association of the antibody's heavy and light chains, each of which has a VH and VL. VH and VL each consist of (1) three complementarity-determining regions (hereinafter sometimes abbreviated as "CDRs") (VH complementarity-determining region 1 (hereinafter sometimes abbreviated as "VH-CDR1"), VH complementarity-determining region 2 (hereinafter sometimes abbreviated as "VH-CDR2"), and VH complementarity-determining region 3 (hereinafter sometimes abbreviated as "VH-CDR3"), as well as VL complementarity-determining region 1 (hereinafter sometimes abbreviated as "VL-CDR1"), VL complementarity-determining region 2 (hereinafter sometimes abbreviated as "VL-CDR2"), and VL complementarity-determining region 3 (hereinafter sometimes abbreviated as "VL-CDR3")) and (2) a framework region (hereinafter sometimes abbreviated as "FW") that positions the CDRs so that the combination of these CDRs can recognize the target antigen or epitope.

[0017] As used herein, the term "heavy chain / light chain complex" refers to a complex consisting of an antibody heavy chain and its corresponding light chain, where "corresponding" refers to a corresponding relationship between the heavy chain and the light chain in which the VH of the heavy chain and the VL of the light chain form an antigen-binding site.

[0018] As used herein, "scFv" is an abbreviation for single-chain Fv, which is a minibody engineered to be produced in a single-chain form so that VH and VL can form an antigen-binding site.

[0019] As used herein, "cross-competition" refers to the mutual inhibition or suppression of binding of two or more antibodies to a common epitope, regardless of the strength of that binding. Cross-competition can be assessed by competitive binding assays, such as Biacore analysis, ELISA assays, flow cytometry, enzyme-linked immunosorbent assay (ELISA), fluorescence energy transfer (FRET), or fluorescence micrometric assay technology (FMAT®).

[0020] [Antigen-binding sites of anti-Allergin-1 bispecific antibodies] The anti-Allergin-1 bispecific antibodies of the present invention are bispecific antibodies having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1. The two antigen-binding sites do not compete with each other for Allergin-1 and are selected from any two of the following four types: (a) an antigen-binding site that specifically binds to a specific epitope on the S1 domain of Allergin-1 (sometimes abbreviated herein as "S1-A binding site"), (b) an antigen-binding site that specifically binds to another epitope on the S1 domain of Allergin-1 that is different from the epitope on the S1 domain described in (a) above (sometimes abbreviated herein as "S1-B binding site"), (c) an antigen-binding site that specifically binds to a specific epitope on the S2 domain of Allergin-1 (sometimes abbreviated herein as "S2-C binding site"), and (d) an antigen-binding site that specifically binds to another epitope on the S2 domain of Allergin-1 that is different from the epitope on the S2 domain described in (c) above (sometimes abbreviated herein as "S2-D binding site"). That is, the S1-A binding site, the S1-B binding site, the S2-C binding site, and the S2-D binding site are antigen-binding sites that do not compete with each other for Allergin-1.

[0021] Here, the epitope to which the S1-A binding site specifically binds (hereinafter, sometimes abbreviated as "S1-A epitope") is at least any of phenylalanine at position 70, arginine at position 71, arginine at position 72, lysine at position 73, histidine at position 75, leucine at position 76, histidine at position 96, glutamic acid at position 97, lysine at position 102, lysine at position 104, and tyrosine at position 113 of Allergin-1 consisting of the amino acid sequence represented by SEQ ID NO: 115. The epitope to which the S2-D binding site specifically binds (hereinafter sometimes abbreviated as "S2-D epitope") is composed of at least one or all of the following amino acids: isoleucine at position 132, methionine at position 133, valine at position 134, isoleucine at position 135, glutamine at position 136, threonine at position 137, glutamic acid at position 138, histidine at position 146, and proline at position 215. On the other hand, the epitope to which the S1-B binding site specifically binds (hereinafter sometimes abbreviated as "S1-B epitope") is an epitope that is different from the S1-A epitope represented by at least the above amino acids, and the epitope to which the S2-C binding site specifically binds (hereinafter sometimes abbreviated as "S2-C epitope") is an epitope that is different from the S2-D epitope represented by at least the above amino acids. Furthermore, "specifically binds" means that the binding is at least 1 × 10 －５ M, preferably 1 × 10 －７ M, more preferably 1×10 －９This means that the S1 or S2 domain of Allergin-1 can bind directly with a binding activity having a higher affinity (dissociation constant (Kd value)) than M, and at least does not substantially bind to other domains of Allergin-1 or other proteins. "Do not compete with each other" means, for example, that the S1-B binding site does not substantially inhibit the binding of the S1-A binding site to its epitope, or that the S1-A binding site does not substantially inhibit the binding of the S1-B binding site to its epitope, or that the S2-D binding site does not substantially inhibit the binding of the S2-C binding site to its epitope, or that the S2-C binding site does not substantially inhibit the binding of the S2-D binding site to its epitope.

[0022] [S1-A binding site and S1-B binding site] One embodiment of the "S1-A binding site" and "S1-B binding site" of the present invention includes, for example, with respect to the S1-A binding site, (A1) an antigen-binding site consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (A2) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30; (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (A3) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (A4) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) an antigen-binding site consisting of a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, while the S1-B binding site is(B1) an antigen-binding site consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63, and (B2) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66.

[0023] Furthermore, the "S1-A binding site" and "S1-B binding site" of the present invention also include those in which any one to five amino acid residues in each CDR constituting each of the VH and VL in (A1) to (A4) and (B1) and (B2) above have been substituted with other amino acids (preferably, conservative amino acids), and which have substantially the same binding activity to each epitope on the S1 domain as that of the original antigen-binding site without the amino acid substitutions. For example, in the case of VH-CDR1 of the S1-A binding site, one amino acid residue has been substituted with the conservative amino acid, and in the case of VH-CDR2 or VH-CDR3, one or two amino acid residues have been substituted with the conservative amino acid. Here, "conservative amino acid" substitution refers to the replacement of residues with similar side chains, such as glycine, alanine, valine, leucine and isoleucine in the group of amino acids with aliphatic side chains, serine and threonine in the group of amino acids with aliphatic hydroxyl side chains, asparagine and glutamine in the group of amino acids with amide-containing side chains, phenylalanine, tyrosine and tryptophan in the group of amino acids with aromatic side chains, lysine, arginine and histidine in the group of amino acids with basic side chains, and cysteine ​​and methionine in the group of amino acids with sulfur-containing side chains.Preferred examples of conservative amino acid substitution include the substitution between valine, leucine and isoleucine, the substitution between phenylalanine and tyrosine, the substitution between lysine and arginine, the substitution between alanine and valine, and the substitution between asparagine and glutamine. Furthermore, the above phrase "having binding activity for each epitope on the S1 domain that is substantially equivalent to the binding activity of the original antigen-binding site that is not substituted with the amino acid" means that the binding activity of the S1-A binding site or S1-B binding site that has been substituted with the amino acid for each epitope on the S1 domain is 95% or more, preferably 98% or more, and more preferably 99% or more of the binding activity of the original S1-A binding site or S1-B binding site that is not substituted with the amino acid.

[0024] Furthermore, the "S1-A binding site" and "S1-B binding site" of the present invention also include those in which the VH and VL constituting them each contain CDRs with the specific amino acid sequences described above, and the FW is humanized so that it is encoded by a specific human germline gene or by such a gene that has undergone one or more somatic mutations and / or backmutations. For example, the FWs encoded by the human germline VH gene IGHV3-66 and VL gene IGKV1-39 or by such a gene that has undergone one or more somatic mutations and / or backmutations can be applied to the VH and VL CDRs shown in (A1) to (A4) and (B1) and (B2) above, respectively. Here, "somatic mutation" refers to a mutation in an antibody gene due to the action of AID (activation-induced cytidine deaminase), and "backmutation" refers to the substitution of one or more amino acid residues in the FW of a humanized antibody with the corresponding amino acid residues in the original non-human (e.g., rabbit, mouse, etc.) antibody. Examples of S1-A binding sites having such humanized VH and VL include antigen-binding sites composed of a VH and a VL consisting of any pair of amino acid sequences selected from: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104; (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105; (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106; (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107; (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108; and (f) SEQ ID NO: 6 and SEQ ID NO: 17 or 109. Examples of S1-B binding sites include antigen-binding sites composed of a VH and a VL consisting of a pair of amino acid sequences: (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110; or (b) SEQ ID NO: 8 and SEQ ID NO: 19 or 111.

[0025] Furthermore, the "S1-A binding site" of the present invention also includes those having VH and VL amino acid sequences that are at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identical to any one pair of amino acid sequences (excluding the three CDRs) selected from the group consisting of: (a) SEQ ID NO:1 and SEQ ID NO:12 or 104; (b) SEQ ID NO:2 and SEQ ID NO:13 or 105; (c) SEQ ID NO:3 and SEQ ID NO:14 or 106; (d) SEQ ID NO:4 and SEQ ID NO:15 or 107; (e) SEQ ID NO:5 and SEQ ID NO:16 or 108; and (f) SEQ ID NO:6 and SEQ ID NO:17 or 109, and wherein differences from the original VH and VL amino acid sequences do not substantially affect the binding activity to the same epitope on the S1 domain (hereinafter sometimes abbreviated as "homologous S1-A binding site"). For example, homologous S1-A binding sites include an S1-A binding site having a VH consisting of the amino acid sequence of SEQ ID NO: 1 or 5, in which the cysteine ​​at position 45 or 44 (corresponding to position 44 according to the Kabat numbering system) is substituted with glycine, respectively, and an S1-A binding site having a VH consisting of an amino acid sequence selected from SEQ ID NOs: 2, 3, 4, and 6, in which the glycine at position 44 (corresponding to position 44 according to the Kabat numbering system) is substituted with cysteine. Here, "% identical" when used in comparing amino acid sequence identity is defined as the percentage of amino acid sequence that is identical to the reference amino acid sequence (after aligning the two sequences and introducing gaps, if necessary, to achieve the maximum percent identity) Furthermore, herein, "differences from the amino acid sequences of the original VH and VL do not substantially affect the binding activity to the same epitope on the S1 domain" means that the binding activity of the homologous S1-A binding site to the same epitope on the S1 domain is 95% or more, preferably 98% or more, and more preferably 99% or more of the binding activity of the S1-A binding site consisting of the original VH and VL.

[0026] Similarly, the "S1-B binding site" of the present invention also includes, for example, a site having a VH and a VL consisting of amino acid sequences that are at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identical to a pair of amino acid sequences (excluding the three CDRs of each) of: (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110; or (b) SEQ ID NO: 8 and SEQ ID NO: 19 or 111, and wherein differences from the original VH and VL amino acid sequences do not substantially affect the binding activity to the same epitope on the S1 domain (hereinafter, sometimes abbreviated as "homologous S1-B binding site"). For example, homologous S1-B binding sites include an S1-B binding site having a VH consisting of the amino acid sequence of SEQ ID NO: 7 in which the cysteine ​​at position 44 (corresponding to position 44 in the Kabat numbering system) is substituted with glycine, and an S1-B binding site having a VH consisting of the amino acid sequence of SEQ ID NO: 8 in which the glycine at position 44 (corresponding to position 44 in the Kabat numbering system) is substituted with cysteine.

[0027] In the present invention, it is preferable that a VH consisting of the amino acid sequence of SEQ ID NO: 1 or 5 and a VL consisting of an amino acid sequence selected from SEQ ID NOs: 12 to 17 each constitute an scFv, while a VH consisting of an amino acid sequence selected from SEQ ID NOs: 2, 3, 4, and 6 and a VL consisting of an amino acid sequence selected from SEQ ID NOs: 104 to 109 each constitute a part of a heavy chain / light chain complex. Also, in the present invention, it is preferable that a VH consisting of the amino acid sequence of SEQ ID NO: 7 and a VL consisting of the amino acid sequence of SEQ ID NO: 18 or 19 each constitute an scFv, while a VH consisting of the amino acid sequence of SEQ ID NO: 8 and a VL consisting of the amino acid sequence of SEQ ID NO: 110 or 111 each constitute a part of a heavy chain / light chain complex.

[0028] In yet another embodiment, the "S1-A binding site" of the present invention includes an antigen-binding site that cross-competes with the binding to an epitope on the S1 domain of an antibody or antigen-binding site thereof having a VH and VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO:1 and SEQ ID NO:104, (b) SEQ ID NO:2 and SEQ ID NO:105, (c) SEQ ID NO:3 and SEQ ID NO:106, (d) SEQ ID NO:4 and SEQ ID NO:107, (e) SEQ ID NO:5 and SEQ ID NO:108, and (f) SEQ ID NO:6 and SEQ ID NO:109, and the binding to the epitope on the S1 domain of the antibody or antigen-binding site thereof cross-competes with the binding to the epitope on the S1 domain of the antibody or antigen-binding site thereof consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO:1 and SEQ ID NO:104, (b) SEQ ID NO:2 and SEQ ID NO:105, (c) SEQ ID NO:3 and SEQ ID NO:106, (d) SEQ ID NO:4 and SEQ ID NO:107, (e) SEQ ID NO:5 and SEQ ID NO:108, and (f) An antigen-binding site that is cross-competed by an antibody or antigen-binding site thereof having VH and VL consisting of any one pair of amino acid sequences selected from SEQ ID NO: 6 and SEQ ID NO: 109.

[0029] Similarly, the "S1-B binding site" of the present invention also includes an antigen-binding site that cross-competes with the binding to an epitope on the S1 domain by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of (a) SEQ ID NO:7 and SEQ ID NO:110, or (b) SEQ ID NO:8 and SEQ ID NO:111, as well as an antigen-binding site whose binding to the epitope on the S1 domain is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of (a) SEQ ID NO:7 and SEQ ID NO:110, or (b) SEQ ID NO:8 and SEQ ID NO:111.

[0030] For example, an antigen-binding site that cross-competes with an S1-A binding site composed of a VH and a VL consisting of the amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 12 or 104, respectively, for binding to the epitope on the S1 domain includes an antigen-binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO: 2 and SEQ ID NO: 13 or 105; (b) SEQ ID NO: 3 and SEQ ID NO: 14 or 106; (c) SEQ ID NO: 4 and SEQ ID NO: 15 or 107; (d) SEQ ID NO: 5 and SEQ ID NO: 16 or 108; and (e) SEQ ID NO: 6 and SEQ ID NO: 17 or 109.

[0031] Furthermore, an antigen-binding site that cross-competes with an S1-B binding site composed of a VH and a VL having the amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 18 or 110, respectively, for binding to the epitope on the S1 domain includes, for example, an antigen-binding site composed of a VH and a VL having the amino acid sequences of SEQ ID NO: 8 and SEQ ID NO: 19 or 111, respectively.

[0032] Here, the "S1-A binding site" of the present invention is preferably an antigen-binding site composed of VH and VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NOs: 1 and 12, (b) SEQ ID NOs: 2 and 13, (c) SEQ ID NOs: 3 and 14 or 106, (d) SEQ ID NOs: 4 and 15 or 107, (e) SEQ ID NOs: 5 and 16, and (f) SEQ ID NOs: 6 and 17 or 109, more preferably: (a) SEQ ID NOs: 3 and 14 or 106, (b) SEQ ID NOs: 4 and 15 or 107, (c) SEQ ID NOs: 5 and 16, and (d) SEQ ID NOs: 6 and 17 or 109, even more preferably: (a) SEQ ID NOs: 3 and 14 or 106, (b) SEQ ID NOs: 4 and 15 or 107, and (c) an antigen-binding site composed of VH and VL consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 5 and 16, and the "S1-B binding site" of the present invention is preferably an antigen-binding site composed of VH and VL consisting of a pair of amino acid sequences: (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110, or (b) SEQ ID NO: 8 and SEQ ID NO: 19 or 111.

[0033] The correspondence between the amino acid sequences of VH and VL constituting the S1-A binding site and the S1-B binding site, respectively, and SEQ ID NOs are shown in Figures 1 and 2. The correspondence between the amino acid sequences of the multiple CDRs contained in each of the VH and VL and SEQ ID NOs is shown in Figures 3 and 4, respectively.

[0034] [S2-C Binding Site and S2-D Binding Site] One embodiment of the "S2-C binding site" and "S2-D binding site" of the present invention includes, for example, an antigen-binding site for the S2-C binding site: (C1) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69; while, for the S2-D binding site: (D1) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48; (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72.

[0035] Furthermore, the "S2-C binding site" and "S2-D binding site" of the present invention also include those in which any one to five amino acid residues in each CDR constituting each of the VH and VL in (C1) and (D1) above have been substituted with other amino acids (preferably, conservative amino acids), and which have substantially the same binding activity to each epitope on the S2 domain as that of the original antigen-binding site without the amino acid substitutions. For example, in the case of VH-CDR1 of the S2-C binding site, one amino acid residue may be substituted with a conservative amino acid, and in the case of VH-CDR2 or VH-CDR3, one or two amino acid residues may be substituted with a conservative amino acid. Here, examples of substitutions with "conservative amino acids" include the amino acid substitutions in the "S1-A binding site" or "S1-B binding site" described above. Furthermore, "having binding activity substantially equivalent to that of the original antigen-binding site not substituted with the amino acid(s) for each epitope on the S2 domain" means, as described above, that the binding activity of the S2-C binding site or S2-D binding site substituted with the amino acid(s) for each epitope on the S2 domain is 95% or more, preferably 98% or more, and more preferably 99% or more of the binding activity of the original S2-C binding site or S2-D binding site not substituted with the amino acid(s).

[0036] Furthermore, the "S2-C binding site" and "S2-D binding site" of the present invention also include those in which the VH and VL constituting them each contain CDRs with the specific amino acid sequences described above, and the FWs are humanized so that they are encoded by a specific human germline gene or by such a gene that has undergone one or more somatic mutations and / or back mutations. For example, as described above, the FWs encoded by the human germline VH gene IGHV3-66 and VL gene IGKV1-39 or by such a gene that has undergone one or more somatic mutations and / or back mutations can be applied to the CDRs of the VH and VL shown in (C1) and (D1) above, respectively. Examples of S2-C binding sites having such humanized VH and VL include antigen-binding sites composed of a VH and VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and examples of S2-D binding sites include antigen-binding sites composed of a VH and VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) SEQ ID NO: 11 and SEQ ID NO: 22 or 114.

[0037] Furthermore, the "S2-C binding site" of the present invention also includes a VH and VL consisting of amino acid sequences that are at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identical to a pair of amino acid sequences (excluding the three CDRs) of SEQ ID NO:9 and SEQ ID NO:20 or 112, respectively, and in which differences from the original VH and VL amino acid sequences do not substantially affect the binding activity to the same epitope on the S2 domain (hereinafter sometimes abbreviated as "homologous S2-C binding site"). For example, a homologous S2-C binding site includes an S2-C binding site having a VH consisting of the amino acid sequence of SEQ ID NO:9 in which the glycine at position 44 (corresponding to position 44 according to the Kabat numbering system) is substituted with a cysteine.

[0038] Similarly, the "S2-D binding site" of the present invention also includes, for example, a VH and VL consisting of amino acid sequences that are at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identical to a pair of amino acid sequences (excluding the three CDRs of each) of: (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113; or (b) SEQ ID NO: 11 and SEQ ID NO: 22 or 114, and wherein differences from the original VH and VL amino acid sequences do not substantially affect the binding activity to the same epitope on the S2 domain (hereinafter sometimes abbreviated as "homologous S2-D binding site"). For example, homologous S2-D binding sites include an S2-D binding site having a VH consisting of the amino acid sequence of SEQ ID NO: 11 in which the cysteine ​​at position 44 (corresponding to position 44 in the Kabat numbering system) is substituted with glycine, and an S2-D binding site having a VH consisting of the amino acid sequence of SEQ ID NO: 10 in which the glycine at position 44 (corresponding to position 44 in the Kabat numbering system) is substituted with cysteine.

[0039] In the present invention, it is preferable that a VL consisting of the amino acid sequence of SEQ ID NO: 20 constitutes an scFv, while it is preferable that a VH consisting of the amino acid sequence of SEQ ID NO: 9 and a VL consisting of the amino acid sequence of SEQ ID NO: 112 each constitute a part of a heavy chain / light chain complex. Also, in the present invention, it is preferable that a VH consisting of the amino acid sequence of SEQ ID NO: 11 and a VL consisting of the amino acid sequence of SEQ ID NO: 21 or 22 each constitute an scFv, while it is preferable that a VH consisting of the amino acid sequence of SEQ ID NO: 10 and a VL consisting of the amino acid sequence of SEQ ID NO: 113 or 114 each constitute a part of a heavy chain / light chain complex.

[0040] In yet another embodiment, the "S2-C binding site" of the present invention also includes an antigen-binding site that cross-competes with an antibody or antigen-binding site thereof having a VH and VL consisting of the amino acid sequences of SEQ ID NO:9 and SEQ ID NO:112, respectively, for binding to an epitope on the S2 domain, as well as an antigen-binding site whose binding to the epitope on the S2 domain is cross-competed with an antibody or antigen-binding site thereof having a VH and VL consisting of a pair of amino acid sequences of SEQ ID NO:9 and SEQ ID NO:112, respectively.

[0041] Similarly, the "S2-D binding site" of the present invention also includes an antigen-binding site that cross-competes with the binding to an epitope on the S2 domain by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of (a) SEQ ID NO: 10 and SEQ ID NO: 113, or (b) SEQ ID NO: 11 and SEQ ID NO: 114, as well as an antigen-binding site whose binding to the epitope on the S2 domain is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of (a) SEQ ID NO: 10 and SEQ ID NO: 113, or (b) SEQ ID NO: 11 and SEQ ID NO: 114.

[0042] For example, an antigen-binding site that cross-competes with an S2-D binding site composed of a VH and a VL consisting of the amino acid sequences of SEQ ID NO: 10 and SEQ ID NO: 21 or 113, respectively, for binding to the epitope on the S2 domain includes, for example, an antigen-binding site composed of a VH and a VL consisting of the amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 22 or 114, respectively.

[0043] Here, the "S2-C binding site" of the present invention is preferably an antigen-binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and the "S2-D binding site" of the present invention is preferably an antigen-binding site composed of: (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 11 and 22.

[0044] The correspondence between the amino acid sequences of VH and VL constituting the S2-C binding site and the S2-D binding site, respectively, and SEQ ID NOs is shown in Figure 5. The correspondence between the amino acid sequences of the multiple CDRs contained in each of the VH and VL and SEQ ID NOs is shown in Figure 6.

[0045] [Form of Anti-Allergin-1 Bispecific Antibody] The anti-Allergin-1 bispecific antibody of the present invention may be in the form of, for example, a diabody (see Proc. Natl. Acad. Sci. USA (1993), Vol. 90, No. 14: pp. 6444-6448), a bispecific sc(Fv) antibody, or the like. ２ (see J. Biological Chemistry (1994), Vol. 269: p.199-206), bispecific F(ab') ２ (see Invest. Ophthalmol. Visual Sci. (2015), Vol. 56: p.5390-5400), bispecific minibody (see Biochemistry (1992), Vo.31, No.6, p.1579-1584), bispecific hybrid antibody, covalent diabody (bispecific DART) (Blood (2011), Vol. 117, No.17, p.4542-4551), bispecific (FvCys) ２ (see J. Immunol. (1992), Vol. 149: p.120-126), bispecific F(ab'-zipper) ２ (See J. Immunol. (1992), Vol. 148, No. 5: pp. 1547-1553), bispecific (Fv-zipper) ２ (Biochemistry (1992), Vol. 31, No. 6: p.1579-1584), bispecific three-chain antibody (Proc. Natl. Acad. Sci. USA (1993), Vol. 90, No. 14: p.6444-6448), bispecific mAb ２ (www.f-star.com / technology_mab.html) and Morrison foam antibodies.

[0046] A bispecific hybrid antibody is an intact antibody in which heavy and light chain complexes of antibodies that recognize two different antigens are covalently linked by disulfide bonds or the like. Bispecific hybrid antibodies can be produced, for example, from hybridomas prepared by the hybrid hybridoma method (see U.S. Pat. No. 4,474,893). Alternatively, bispecific hybrid antibodies can be produced by coexpressing and secreting four types of cDNAs, each encoding the heavy and light chains of antibodies that recognize different antigens, in mammalian cells.

[0047] Morrison-form antibodies are bispecific antibodies in which an scFv that binds to an epitope different from that of the antibody is linked to the C-terminus of the heavy chain or the C-terminus of the light chain of the antibody (see Nat. Biotechnol. (1997), Vol. 15, No. 2: pp. 159-63 and Protein Eng. Des. Sel. (2010), Vol. 23, No. 4: pp. 221-228).

[0048] The anti-Allergin-1 bispecific antibody of the present invention is preferably a Morrison foam antibody.

[0049] When the anti-Allergin-1 bispecific antibody of the present invention is a Morrison form antibody, one antigen-binding site of the bispecific antibody is formed as part of the heavy chain / light chain complex of the bispecific antibody, and the other antigen-binding site is in the form of an scFv consisting of the VH and VL constituting the heavy chain / light chain complex. For example, the heavy chain / light chain complex may be formed by any one combination selected from the following: (a) a combination of a heavy chain having the VH constituting the S1-A binding site and a light chain having the VL constituting the S1-B binding site, (b) a combination of a heavy chain having the VH constituting the S1-B binding site and a light chain having the VL constituting the S1-B binding site, (c) a combination of a heavy chain having the VH constituting the S2-C binding site and a light chain having the VL constituting the S2-C binding site, and (d) a combination of a heavy chain having the VH constituting the S2-D binding site and a light chain having the VL constituting the S2-D binding site. Here, both the heavy chain and light chain can be those of an IgG antibody, in which case the heavy chain has heavy chain constant region 1 (hereinafter sometimes abbreviated as "CH1"), heavy chain constant region 2 (hereinafter sometimes abbreviated as "CH2"), and heavy chain constant region 3 (hereinafter sometimes abbreviated as "CH3") of an IgG antibody, and the light chain has the light chain constant region (hereinafter sometimes abbreviated as "CL") of an IgG antibody.

[0050] On the other hand, the scFv is selected from antigen-binding sites that are different from and do not compete with the antigen-binding site that constitutes a part of the heavy chain / light chain complex (hereinafter sometimes abbreviated as "antigen-binding site in the heavy chain / light chain complex"). For example, (a) when the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site, it can be selected from any of an S1-B binding site, an S2-C binding site, and an S2-D binding site; (b) when the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site, it can be selected from any of an S1-A binding site, an S2-C binding site, and an S2-D binding site; (c) when the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, it can be selected from any of an S1-A binding site, an S1-B binding site, and an S2-D binding site; or (d) When the antigen-binding site in the heavy chain / light chain complex is an S2-D binding site, it can be selected from an S1-A binding site, an S1-B binding site, and an S2-C binding site.

[0051] Here, the combination of the antigen-binding site in the heavy chain / light chain complex and the antigen-binding site constituting the scFv is preferably a combination of antigen-binding sites that bind to the S1 domain and S2 domain of Allergin-1, respectively. For example, when the antigen-binding site in the heavy chain / light chain complex is the S1-A binding site or the S1-B binding site, the antigen-binding site of the scFv is the S2-C binding site or the S2-D binding site; when the antigen-binding site in the heavy chain / light chain complex is the S2-C binding site or the S2-D binding site, the antigen-binding site of the scFv is the S1-A binding site or the S1-B binding site. Furthermore, when the antigen-binding site in the heavy chain / light chain complex is the S1-A binding site, a more preferred combination is when the antigen-binding site of the scFv is the S2-C binding site or the S2-D binding site; and when the antigen-binding site in the heavy chain / light chain complex is the S2-C binding site or the S2-D binding site, a more preferred combination is when the antigen-binding site in the heavy chain / light chain complex is the S2-C binding site or the S2-D binding site.

[0052] Furthermore, the scFv is linked to the C-terminus of either the heavy chain or the light chain. For example, the N-terminus of the scFv may be linked to the C-terminus of either the heavy chain or the light chain via a direct amide bond, or via a peptide linker. The scFv is preferably linked to the C-terminus of the heavy chain, and more preferably linked to that C-terminus via a peptide linker. Here, "linked via a peptide linker" means that the N-terminus of the peptide linker is linked to the C-terminus of either the heavy chain or the light chain via an amide bond, and further, that C-terminus is linked to the N-terminus of the scFv via an amide bond. In one embodiment, the heavy chain or light chain linked to an scFv that constitutes the anti-Allergin-1 bispecific antibody of the present invention is produced as an integrated protein from a host cell that expresses them.

[0053] The VH and VL constituting the scFv may be linked to each other via a direct amide bond or a peptide linker in the order of VH and VL from the N-terminus, or conversely, they may be linked to each other via a direct amide bond or a peptide linker in the order of VL and VH from the N-terminus, but preferably they are linked to each other via a peptide linker in the order of VL and VH from the N-terminus of the scFv.

[0054] The peptide linker linking the scFv to the C-terminus of either the heavy chain or the light chain may be any peptide linker as long as it exhibits the activity expected of the bispecific antibody of the present invention, but a preferred peptide linker is one having the amino acid sequence Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 73). On the other hand, the peptide linker linking the VH and VL that form the scFv to each other may be any peptide linker as long as it does not impair the antigen-binding ability of the scFv, but a preferred peptide linker is one having the amino acid sequence Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 74). Herein, "Gly" represents glycine, and "Ser" represents serine.

[0055] When the anti-Allergin-1 bispecific antibody of the present invention is a Morrison form antibody, its preferred form is one in which two identical heavy chain / light chain complexes composed of the heavy chain and light chain to which the scFv is linked at the C-terminus of one of the heavy chains in the above-mentioned manner are associated, for example, via a disulfide bond, and such a form is exemplified in Figures 7(A) and (B). The anti-Allergin-1 bispecific antibody of the present invention may be derived from any of chimeric, humanized, and human antibodies.

[0056] Chimeric antibodies can be produced, for example, by ligating genes encoding antibody variable regions isolated by known techniques from antibody-producing hybridomas isolated by the hybridoma method (see, for example, Nature (1975), Vol. 256: pp. 495-97; Hongo et al., Hybridoma (1995), Vol. 14, No. 3, pp. 253-260; Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press (1988), Vol. 2); or Monoclonal Antibodies and T-Cell Hybridomas, pp. 563-681 (Elsevier, NY, 1981)) to genes encoding human-derived antibody constant regions using known methods (see, for example, US Pat. No. 4,816,567).

[0057] Furthermore, in the case of humanized antibodies, genes encoding the CDRs of antibodies isolated by known techniques from antibody-producing hybridomas isolated by the above-described method can be linked to genes encoding human-derived antibody framework regions using known methods to produce humanized antibodies (see, for example, US Pat. No. 5,225,539, US Pat. No. 5,530,101, US Pat. No. 5,585,089, or US Pat. No. 6,180,370).

[0058] On the other hand, a human antibody refers to an antibody whose entire composition is derived from human germline immunoglobulin sequences, and can be produced by a method using a mouse transformed to produce a human antibody, for example, a Humab mouse (see US5545806, US5569825, US5625126, US5633425, US5789650, US5877397, US5661016, US5814318, US5874299, or US5770429), a KM mouse (see WO2002 / 43478), a Xeno mouse (see, for example, US5939598, US6075181, US6114598, US6150584, or US6162963), or a Tc mouse (see Proc. Natl. Acad. Sci. USA (2000), pp. 722-727). They can also be generated using SCID mice reconstituted with human immune cells to generate a human antibody response upon immunization (see US5476996 or US5698767), or by phage display methods (see US5223409, US5403484, US5571698, US5427908, US5580717, US5969108, US6172197, US5885793, US6521404, US6544731, US6555313, US6582915, or US6593081).

[0059] The anti-Allergin-1 bispecific antibody of the present invention may be an antibody of any isotype, but the preferred isotype is IgG, and more preferably IgG. １ or IgG ４ where the preferred IgG １Examples of antibodies that have lost or weakened Fc receptor binding include those that can be obtained by substituting, deleting, or inserting any amino acid in the heavy chain constant region. Examples include antibodies in which leucine at position 235 according to the EU numbering system is substituted with glycine and / or glycine at position 236 is substituted with arginine in the two heavy chain constant regions or hinge regions of a bispecific antibody. Furthermore, to reduce antibody heterogeneity, IgGs lacking the C-terminal amino acid, for example, lysine at position 447 according to the EU numbering system, can be used. １ is more preferred.

[0060] The anti-Allergin-1 bispecific antibody of the present invention is IgG ４ For antibody isotypes of the above, variants in which any amino acid in the heavy chain constant region is substituted, deleted, or inserted to suppress swapping within the antibody molecule are more preferred. For example, an antibody in which serine at position 228 according to the EU numbering system, located in the hinge region, is substituted with proline is preferred. Herein, the amino acid positions assigned to the CDRs and frameworks of antibody variable regions can be defined according to Kabat (see Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md.), (1987) and (1991)), and the amino acids in the constant region can be represented according to the EU numbering system based on Kabat amino acid positions (see Sequences of proteins of immunological interest, NIH Publication No. 91-3242).

[0061] Preferred isotypes for the anti-Allergin-1 bispecific antibodies of the present invention include IgG １ The isotype of the anti-Allergin-1 bispecific antibody of the present invention is IgG １In one embodiment, for example, the antibody has a heavy chain constant region consisting of the amino acid sequence of SEQ ID NO: 23 or 99 (in which the arginine at position 97 (corresponding to position 214 in the heavy chain constant region according to the EU numbering system) in the amino acid sequence of SEQ ID NO: 23 or 99 may be substituted with lysine), and a light chain constant region consisting of the amino acid sequence of SEQ ID NO: 24. The amino acid sequences are shown in Figure 8.

[0062] The "antibody fragment" of the anti-Allergin-1 bispecific antibody of the present invention is a part of the bispecific antibody and has at least one antigen-binding site that specifically binds to Allergin-1. One embodiment of the antibody fragment is an antibody fragment consisting of VH and VL that constitute any of the S1-A binding site, S1-B binding site, S2-C binding site, and S2-D binding site, and examples thereof include antibody fragments consisting of VH and VL exemplified above in the sections "S1-A binding site and S1-B binding site" and "S2-C binding site and S2-D binding site." Examples of antibody fragments consisting of VH and VL include Fab, Fab', Fv, and F(ab'). ２ Furthermore, examples thereof include heavy chain / light chain complexes containing such VH and VL, and complexes in which an scFv consisting of VH and VL constituting the other antigen-binding site is linked to the C-terminus of either the heavy chain or the light chain of the heavy chain / light chain complex. The scFv itself is also included in the antibody fragment.

[0063] In one embodiment, when the anti-Allergin-1 bispecific antibody of the present invention is a Morrison form antibody, it is, for example, a bispecific antibody comprising two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, which each specifically bind to two different epitopes in the extracellular domain of Allergin-1 and do not compete with each other for Allergin-1, wherein: (I) the S1-A binding site is an antigen-binding site comprising: (i) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (ii) an antigen-binding site consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (iii) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57, and (iv) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58,(e) an antigen-binding site selected from the group consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, wherein the S1-B binding site is an antigen-binding site selected from the group consisting of: (v) an VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63, or (vi) an VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, wherein the S2-C binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, wherein the S2-D binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72,(II) One of the two different antigen-binding sites constitutes the antigen-binding site in the heavy chain / light chain complex constituting the bispecific antibody, and the other constitutes an antigen-binding site in the form of an scFv, (i) the antigen-binding site in the heavy chain / light chain complex constituting the bispecific antibody is an S1-A binding site, and the antigen-binding site in the form of an scFv is any one type of antigen-binding site selected from an S1-B binding site, an S2-C binding site, and an S2-D binding site, or (ii) the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site, and the antigen-binding site in the form of an scFv is any one type of antigen-binding site selected from an S1-A binding site, an S2-C binding site, and an S2-D binding site, (iii) the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, and the antigen-binding site in the form of the scFv is any one type of antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-D binding site, or (iv) the antigen-binding site in the heavy chain / light chain complex is an S2-D binding site, and the antigen-binding site in the form of the scFv is any one type of antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-C binding site, (III) the N-terminus of VH or VL constituting the scFv is linked to the C-terminus of either the heavy chain or the light chain via a peptide linker, and (IV) (i) Bispecific antibodies having heavy and light chains with the configurations described in the preceding paragraphs (I) to (III), or (ii) bispecific antibodies consisting of the same two heavy chain / light chain complexes composed of heavy and light chains with the configurations described in the preceding paragraphs (I) to (III).

[0064] Here, preferred combinations of two different antigen-binding sites constituting the anti-Allergin-1 bispecific antibody of the present invention include: (i) an S1-A binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from (a) SEQ ID NOs: 1 and 12, (b) SEQ ID NOs: 2 and 13, (c) SEQ ID NOs: 3 and 14 or 106, (d) SEQ ID NOs: 4 and 15 or 107, (e) SEQ ID NOs: 5 and 16, and (f) SEQ ID NOs: 6 and 17 or 109; (ii) an S1-B binding site composed of a VH and a VL consisting of a pair of amino acid sequences of (a) SEQ ID NOs: 7 and 18 or (b) SEQ ID NOs: 8 and 19; (iii) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 9 and 20 or 112; (iv) any two combinations selected from (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) SEQ ID NO: 11 and 22, and more preferably (i) an S1-A binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from (a) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (b) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (c) SEQ ID NO: 5 and 16, and (d) SEQ ID NO: 6 and SEQ ID NO: 17 or 109, (ii) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and (iii) any two combinations selected from (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) SEQ ID NO: 11 and 22,

[0065] Furthermore, preferred combinations of two different antigen-binding sites constituting the anti-Allergin-1 bispecific antibody of the present invention include: (i) (a) an S1-A binding site composed of a VH and a VL consisting of the pair of amino acid sequences of SEQ ID NO: 3 and SEQ ID NO: 14 or 106, and (b) an S2-C binding site composed of a VH and a VL consisting of the pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112; (ii) (a) an S1-A binding site composed of a VH and a VL consisting of the pair of amino acid sequences of SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and (b) an S2-D binding site composed of a VH and a VL consisting of the pair of amino acid sequences of SEQ ID NO: 10 and SEQ ID NO: 21 or 113; and (iii) (a) (a1) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (a2) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, and (a3) an S1-A binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 5 and 16, and (b) (b1) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 10 and 21 or 113, or (b2) a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 11 and 22, more preferably: (i) a combination of (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 3 and 14, and (b) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 9 and 112, (ii) a combination of (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 3 and 106, and (b) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 9 and 20, (iii) (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 6 and 17, and (b) a combination with an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 10 and 113; (iv) (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 6 and 109, and(b) a combination with an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 21; (v) a combination with (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 14, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113; (vi) a combination with (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 15, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113; (vii) a combination with (a) an S1-A binding site composed of a VH and a VL consisting of any pair of amino acid sequences selected from SEQ ID NOs: 5 and 16, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113; (viii) (a) a combination of an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 106, and (b) a combination of an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 21, and (ix) a combination of (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 107, and (b) a combination of an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 11 and 22.

[0066] Furthermore, the VH and VL constituting the scFv are preferably linked via a peptide linker consisting of the amino acid sequence of SEQ ID NO: 74 in the order of VL and VH from their N-terminus, and the peptide linker linking the scFv and the C-terminus of the heavy chain or light chain is preferably a peptide linker consisting of the amino acid sequence of SEQ ID NO: 73.

[0067] Preferred examples of the anti-Allergin-1 bispecific antibodies of the present invention having an scFv linked to the C-terminus of their heavy chains include: (i) (a) SEQ ID NOs: 75 and 76, (b) SEQ ID NOs: 77 and 78, (c) SEQ ID NOs: 79 and 78, (d) SEQ ID NOs: 80 and 78, (e) SEQ ID NOs: 81 and 82, (f) SEQ ID NOs: 83 and 82, (g) SEQ ID NOs: 84 and 85, (h) SEQ ID NOs: 86 and 87, (i) SEQ ID NOs: 98 and 78, (j) SEQ ID NOs: 100 and 78, (k) SEQ ID NOs: 101 and 78, (l) SEQ ID NOs: 102 and 78, and (m) antibodies having heavy and light chains consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 103 and 85, or (ii) antibodies consisting of the same two heavy chain / light chain complexes consisting of the heavy and light chains described in (i) above. On the other hand, preferred anti-Allergin-1 bispecific antibodies of the present invention having an scFv linked to the C-terminus of their light chains include, for example, antibodies having heavy and light chains consisting of any one pair of amino acid sequences selected from (a) SEQ ID NOs: 88 and 89, (b) SEQ ID NOs: 90 and 91, (c) SEQ ID NOs: 90 and 92, (d) SEQ ID NOs: 93 and 94, (e) SEQ ID NOs: 93 and 95, and (f) SEQ ID NOs: 96 and 97, or (ii) antibodies consisting of the same two heavy chain / light chain complexes consisting of the heavy and light chains described in (i) above. In this regard, the arginine at position 214 according to the EU numbering system in the heavy chain constant region of these anti-Allergin-1 bispecific antibodies may be substituted with lysine, and / or the alanine at position 299 in SEQ ID NOs: 75, 81, 83, 86, 88, 93 and 96 and position 301 in SEQ ID NOs: 80 and 90 (corresponding to position 297 according to the EU numbering system in the heavy chain constant region) may each independently be substituted with asparagine.

[0068] More preferably, the anti-Allergin-1 bispecific antibody of the present invention is an antibody that has heavy and light chains consisting of any one pair of amino acid sequences selected from (a) SEQ ID NOs: 77 and 78, (b) SEQ ID NOs: 79 and 78, (c) SEQ ID NOs: 83 and 82, (d) SEQ ID NOs: 84 and 85, (e) SEQ ID NOs: 98 and 78, (f) SEQ ID NOs: 100 and 78, (g) SEQ ID NOs: 101 and 78, (h) SEQ ID NOs: 102 and 78, and (j) SEQ ID NOs: 103 and 85, or (ii) an antibody that consists of the same two heavy chain / light chain complexes consisting of the heavy chain and light chain described in (i) above.

[0069] A preferred feature of the anti-Allergin-1 bispecific antibodies of the present invention is that cytokine production is sufficiently reduced during administration or within 24 hours after administration. Here, "sufficiently reduced cytokine production" means, for example, that during intravenous administration of the anti-Allergin-1 bispecific antibodies of the present invention by infusion or the like, or within 24 hours after administration, the concentrations of cytokines, including at least one selected from IL-2, IL-4, IL-6, IL-10, IFN-γ, and TNF-α, in blood or tissues do not increase, or, if they increase, they are at a level that can be suppressed by steroid administration.

[0070] One aspect of the characteristics of the anti-Allergin-1 bispecific antibodies of the present invention is that they inhibit the binding of a TLR9 ligand (e.g., an oligodeoxynucleotide (ODN), in one embodiment, a CpG ODN, preferably ODN M362, ODN 1668, or ODN 1826) to Allergin-1.

[0071] [Methods for producing and purifying anti-Allergin-1 bispecific antibodies] The anti-Allergin-1 bispecific antibodies of the present invention and antibody fragments thereof can be produced by secreting them from host cells according to known methods. For example, they can be produced by transfecting mammalian cells with expression vectors containing (1) a polynucleotide encoding a heavy chain linked to an scFv at its C-terminus and (2) a polynucleotide encoding a light chain, and then expressing and secreting both the heavy and light chains.

[0072] Preferred host cells for expressing the anti-Allergin-1 bispecific antibodies of the present invention include insect cells such as SF-9 cells and SF-21 cells, and more preferably (i) mouse cells including CHO cells, BHK cells, SP2 / 0 cells, and NS-0 myeloma cells, (ii) primate cells such as COS cells and Vero cells, and (iii) mammalian cells such as MDCK cells, BRL 3A cells, hybridomas, tumor cells, immortalized primary cells, W138 cells, HepG2 cells, HeLa cells, HEK293 cells, HT1080 cells, and embryonic retinal cells such as PER.C6 cells. In selecting an expression system, mammalian cells and expression vectors therefor may be used to ensure appropriate glycosylation of the antibody. Human cells, preferably PER.C6 cells, are advantageously used to obtain antibodies with glycosylation patterns consistent with those in humans.

[0073] Protein production in host cells transformed by gene introduction of an expression vector can be carried out with reference to, for example, Current Protocols in Protein Science (1995), Coligan JE, Dunn BM, Ploegh HL, Speicher DW, Wingfield PT, ISBN 0-471-11184-8, Bendig, 1988. Furthermore, general guidelines, procedures, and practical methods for maximizing the productivity of host cell culture can be carried out with reference to Mammalian Cell Biotechnology: a Practical Approach (M. Butler, ed., IRL Press, 1991). Expression of antibodies in host cells is described in publications such as EP0120694, EP0314161, EP0481790, EP0523949, US4816567, and WO2000 / 63403.

[0074] Here, the culture conditions of the host cells can be optimized by known methods, thereby optimizing the protein production amount. Culture can be carried out, for example, in a culture dish, roller bottle, or reactor, by batch culture, fed-batch culture, continuous culture, or hollow fiber culture. For large-scale, continuous production of recombinant proteins by cell culture, it is preferable to grow the cells in suspension. It is also preferable to culture the cells in the absence of animal- or human-derived serum or serum components thereof.

[0075] Antibodies expressed in host cells and recovered from the cells or cell culture medium by known methods can be purified using known methods, including immunoprecipitation, centrifugation, filtration, size exclusion chromatography, affinity chromatography, cation and / or anion exchange chromatography, hydrophobic interaction chromatography, etc. In addition, Protein A or Protein G affinity chromatography may be suitably used (see, e.g., US4801687 and US5151504).

[0076] [Anti-Allergin-1 antibodies] The present invention also includes antibodies that specifically bind to the S1 domain of Allergin-1 (hereinafter sometimes simply referred to as "anti-S1 domain antibodies") and antibodies that specifically bind to the S2 domain of Allergin-1 (hereinafter sometimes simply referred to as "anti-S2 domain antibodies") (these may collectively be abbreviated as "anti-Allergin-1 antibodies"), which are used to construct the anti-Allergin-1 bispecific antibodies of the present invention, as well as antibody fragments thereof.

[0077] The anti-S1 domain antibody of the present invention may be any antibody that specifically binds to the S1 domain of Allergin-1 and can be used to construct the anti-Allergin-1 bispecific antibody of the present invention. However, it is preferable to use at least 1 × 10 －５ M, preferably 1 × 10 －７ M, more preferably 1×10 －９ These antibodies can directly bind to the S1 domain of Allergin-1 with a binding activity having an affinity (Kd value) higher than M, and do not substantially bind to at least the S2 domain of Allergin-1 or other proteins. Note that anti-S1 domain antibodies include antibodies that specifically bind to the S1-A epitope (an epitope specifically bound by the S1-A binding site) or the S1-B epitope (an epitope specifically bound by the S1-B binding site).

[0078] On the other hand, the anti-S2 domain antibody of the present invention may be any antibody that specifically binds to the S2 domain of Allergin-1 and can be used to construct the anti-Allergin-1 bispecific antibody of the present invention. －５ M, preferably 1 × 10 －７ M, more preferably 1×10 －９These antibodies can directly bind to the S2 domain of Allergin-1 with a binding activity having an affinity (Kd value) higher than M, and at least do not substantially bind to the S1 domain of Allergin-1 or other proteins. Anti-S2 domain antibodies include antibodies that specifically bind to the S2-C epitope (the epitope to which the S2-C binding site specifically binds) or the S2-D epitope (the epitope to which the S2-D binding site specifically binds). Here, the term "antibody" in "anti-Allergin-1 antibody," "anti-S1 domain antibody," and "anti-S2 domain antibody" refers to a full-length antibody, i.e., a full-length antibody consisting of two heavy chains and two light chains. Their "antibody fragments" refer to antibody portions that are part of a full-length antibody and contain at least the antigen-binding portion, such as Fab, Fab', Fv, and F(ab'). ２ Examples include:

[0079] The anti-Allergin-1 antibodies of the present invention are preferably chimeric, humanized, or human antibodies in constructing the anti-Allergin-1 bispecific antibodies of the present invention, but also include non-human antibodies (e.g., mouse, rat, and rabbit antibodies) used in producing chimeric or humanized antibodies.

[0080] One embodiment of the anti-S1 domain antibody of the present invention includes, for example: (E1) an antibody consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (E2) an antibody consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) an antibody consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (E3) (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) an antibody consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; (E4) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59; (f) an antibody consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60; (E5) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37; (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38; and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61;(e) an antibody consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; and (E6) an antibody consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66. On the other hand, one embodiment of the anti-S2 domain antibody includes, for example: (F1) an antibody consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69; and (F2) an VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) An antibody consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72.

[0081] Furthermore, the anti-Allergin-1 antibodies of the present invention also include antibodies in which any one to five amino acid residues in each CDR have been substituted with other amino acids (preferably conservative amino acids) and which have substantially the same binding activity to the same epitope as the original antibody without the amino acid substitutions. For example, in the case of CDR1, one amino acid residue has been substituted with a conservative amino acid, and in the case of CDR2 or CDR3, one or two amino acid residues have been substituted with conservative amino acids.

[0082] Furthermore, the anti-Allergin-1 antibodies of the present invention also include those whose VH and VL contain CDRs consisting of the above-mentioned specific amino acid sequences, and whose framework region amino acid sequences are encoded by specific germline genes or by such genes that have undergone one or more somatic mutations and / or back mutations. Examples include VH and VL encoded by the above-mentioned germline VH gene IGHV3-66 and VL gene IGKV1-39, or by such genes that have undergone one or more somatic mutations and / or back mutations.

[0083] Further, other embodiments of the anti-S1 domain antibodies of the present invention include antibodies having a VH and VL consisting of any one pair of amino acid sequences selected from, for example, (G1) SEQ ID NOs: 1 and 104, (G2) SEQ ID NOs: 2 and 105, (G3) SEQ ID NOs: 3 and 106, (G4) SEQ ID NOs: 4 and 107, (G5) SEQ ID NOs: 5 and 108, (G6) SEQ ID NOs: 6 and 109, (G7) SEQ ID NOs: 7 and 110, and (G8) SEQ ID NOs: 8 and 111. Meanwhile, other embodiments of the anti-S2 domain antibodies include antibodies having a VH and VL consisting of any one pair of amino acid sequences selected from, for example, (H1) SEQ ID NOs: 9 and 112, (H2) SEQ ID NOs: 10 and 113, and (H3) SEQ ID NOs: 11 and 114.

[0084] Furthermore, in another such embodiment, anti-Allergin-1 antibodies also include antibodies having VH and VL sequences that are independently at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identical to any one set of amino acid sequences selected from the aforementioned (G1) to (G8) (excluding each of the three CDRs) or any one set of amino acid sequences selected from the aforementioned (H1) to (H3) (excluding each of the three CDRs), and in which differences from the original VH and VL amino acid sequences do not substantially affect the binding activity to the same epitope.

[0085] Furthermore, another embodiment of the anti-S1 domain antibodies of the present invention includes, for example, antibodies that cross-compete for binding to an epitope on the S1 domain by an antibody having a VH and VL consisting of any one pair of amino acid sequences selected from the above-mentioned (G1) to (G8), or an antigen-binding site thereof, as well as antibodies whose binding to the epitope on the S1 domain is cross-competed by an antibody having a VH and VL consisting of any one pair of amino acid sequences selected from the above-mentioned (G1) to (G8), or an antigen-binding site thereof.

[0086] Another embodiment of the anti-S2 domain antibody also includes, for example, an antibody whose binding to an epitope on the S2 domain is cross-competed with an antibody having a VH and a VL consisting of any one pair of amino acid sequences selected from the above-mentioned (H1) to (H3), or its antigen-binding site, as well as an antibody whose binding to the epitope on the S2 domain is cross-competed with an antibody having a VH and a VL consisting of any one pair of amino acid sequences selected from the above-mentioned (H1) to (H3), or its antigen-binding site.

[0087] The anti-Allergin-1 antibody of the present invention is preferably a monoclonal antibody. Monoclonal antibodies can be obtained by various known methods, including the hybridoma method described above. Preferred isotypes for the anti-Allergin-1 antibody of the present invention include IgG. １ is.

[0088] The isotype of the anti-Allergin-1 antibody of the present invention is IgG １ In one embodiment, for example, the antibody has a heavy chain constant region consisting of the amino acid sequence of SEQ ID NO: 23 or 99 (in which the arginine at position 97 (corresponding to position 214 in the heavy chain constant region according to the EU numbering system) in the amino acid sequence of SEQ ID NO: 23 or 99 may be substituted with lysine), and a light chain constant region consisting of the amino acid sequence of SEQ ID NO: 24.

[0089] [Polynucleotides Encoding Anti-Allergin-1 Bispecific Antibodies] The present invention also includes polynucleotides encoding each portion of the anti-Allergin-1 bispecific antibodies of the present invention used in producing the bispecific antibodies, i.e., the VH and VL that each constitute any two of the S1-A binding site, S1-B binding site, S2-C binding site, and S2-D binding site, as well as the heavy chain constant region and light chain constant region.

[0090] Examples of polynucleotides encoding the VH and VL that constitute the S1-A binding site include polynucleotides that encode VH and VL, respectively, consisting of any pair of amino acid sequences selected from (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) SEQ ID NO: 6 and SEQ ID NO: 17 or 109. Examples of polynucleotides encoding the VH and VL that constitute the S1-B binding site include polynucleotides that encode VH and VL, respectively, consisting of the pair of amino acid sequences set forth in (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110, or (b) SEQ ID NO: 8 and SEQ ID NO: 19 or 111.

[0091] On the other hand, examples of polynucleotides encoding the VH and VL that constitute the S2-C binding site include polynucleotides that encode VH and VL, respectively, consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and examples of polynucleotides encoding VH and VL that constitute the S2-D binding site include polynucleotides that encode VH and VL, respectively, consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) SEQ ID NO: 11 and SEQ ID NO: 22 or 114.

[0092] Examples of polynucleotides encoding the heavy chain constant region and the light chain constant region, respectively, include polynucleotides encoding the amino acid sequences of SEQ ID NO: 23 or 99 and SEQ ID NO: 24, respectively.

[0093] When the C-terminus of the heavy chain or light chain and the scFv, and the VH and VL constituting the scFv are linked via peptide linkers, polynucleotides encoding these peptide linkers are inserted between the polynucleotides encoding the heavy chain or light chain and the scFv or the VH and VL constituting the scFv, respectively. An example of a polynucleotide encoding the former peptide linker is a polynucleotide encoding the amino acid sequence of SEQ ID NO: 73, and an example of a polynucleotide encoding the latter peptide linker is a polynucleotide encoding the amino acid sequence of SEQ ID NO: 74.

[0094] Preferred examples of polynucleotides encoding anti-Allergin-1 bispecific antibodies of the present invention having an scFv linked to the C-terminus of their heavy chains include polynucleotides encoding heavy and light chains consisting of any one set of amino acid sequences selected from: (a) SEQ ID NOs: 75 and 76, (b) SEQ ID NOs: 77 and 78, (c) SEQ ID NOs: 79 and 78, (d) SEQ ID NOs: 80 and 78, (e) SEQ ID NOs: 81 and 82, (f) SEQ ID NOs: 83 and 82, (g) SEQ ID NOs: 84 and 85, (h) SEQ ID NOs: 86 and 87, (i) SEQ ID NOs: 98 and 78, (j) SEQ ID NOs: 100 and 78, (k) SEQ ID NOs: 101 and 78, (l) SEQ ID NOs: 102 and 78, and (m) SEQ ID NOs: 103 and 85. Preferred examples of polynucleotides encoding anti-Allergin-1 bispecific antibodies of the present invention having an scFv linked to the C-terminus of their light chains include: Examples of such polynucleotides include polynucleotides encoding heavy and light chains each consisting of any pair of amino acid sequences selected from: (a) SEQ ID NOs: 88 and 89, (b) SEQ ID NOs: 90 and 91, (c) SEQ ID NOs: 90 and 92, (d) SEQ ID NOs: 93 and 94, (e) SEQ ID NOs: 93 and 95, and (f) SEQ ID NOs: 96 and 97. As described above, in the heavy chain constant region of the anti-Allergin-1 bispecific antibody of the present invention, arginine at position 214 according to the EU numbering system may be substituted with lysine, and / or alanine at position 299 in SEQ ID NOs: 75, 81, 83, 86, 88, 93, and 96 and alanine at position 301 in SEQ ID NOs: 80 and 90 (corresponding to position 297 according to the EU numbering system in the heavy chain constant region) may each be independently substituted with asparagine.

[0095] Here, 1 to 6 types of codons encoding one amino acid are known, and for example, TTT or TTC for Phe, TTA, TTG, CTT, CTC, CTA or CTG for Leu, ATT, ATC or ATA for Ile, ATG for Met, GTT, GTC, GTA or GTG for Val, TCT, TCC, TCA or TCG for Ser, CCT, CCC, CCA or CCG for Pro, ACT, ACC, ACA or ACG for Thr, GCT, GCC, GCA or GCG for Ala, TAT or TAC for Tyr, CAT or CAC for His, and CAT or CAC for Gln. Codons corresponding to each amino acid correspond to CAA or CAG, Asn to AAT or AAC, Lys to AAA or AAG, Asp to GAT or GAC, Glu to GAA or GAG, Cys to TGT or TGC, Trp to TGG, Arg to CGT, CGC, CGA or CGG, Ser to AGT or AGC, Arg to AGA or AGG, and Gly to GGT, GGC, GGA or GGG. Thus, polynucleotides encoding each portion of an anti-Allergin-1 bispecific antibody include polynucleotides in which any combination of codons corresponding to each amino acid is used.

[0096] These polynucleotides include both DNA and RNA forms.

[0097] [Medicinal Uses] Cancers to which the anti-Allergin-1 bispecific antibodies of the present invention can be applied are not particularly limited, and include any solid cancer and blood cancer. Here, examples of solid cancers include malignant melanoma (e.g., malignant melanoma of the skin, oral mucosal epithelium, or orbit, etc.), non-small cell lung cancer (e.g., squamous non-small cell lung cancer and non-squamous non-small cell lung cancer), small cell lung cancer, head and neck cancer (e.g., oral cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, salivary gland cancer, and tongue cancer), renal cell carcinoma (e.g., clear cell renal cell carcinoma), breast cancer, ovarian cancer (e.g., serous ovarian cancer and ovarian clear cell adenocarcinoma), uterine cancer (e.g., cervical cancer and endometrial cancer), anal cancer (e.g., anal canal cancer), and colorectal cancer (e.g., microsatellite instability-high (hereinafter abbreviated as "MSI-H") and / or mismatch repair deficient (hereinafter abbreviated as "dMMR") positive). colorectal cancer), rectal cancer, colon cancer, hepatocellular carcinoma, esophageal cancer, gastric cancer, esophagogastric junction cancer, pancreatic cancer (e.g., pancreatic ductal carcinoma, insulinoma, intraductal papillary mucinous neoplasm), urothelial cancer (e.g., bladder cancer, upper urinary tract cancer, ureteral cancer, renal pelvic cancer, and urethral cancer), prostate cancer, fallopian tube cancer, primary peritoneal cancer, malignant pleural mesothelioma, gallbladder cancer, bile duct cancer, biliary tract cancer, skin cancer (e.g., uveal melanoma and Merkel cell carcinoma), testicular cancer (germ cell tumor), vaginal cancer, vulvar cancer, penile cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, spinal tumor, neuroblastoma, medulloblastoma, ocular retinoblastoma, neuroendocrine tumor, brain tumor (e.g., glioma (e.g., glioblastoma and gliosarcoma) and meningioma), and squamous cell carcinoma.

[0098] Among solid cancers, sarcomas include bone and soft tissue sarcomas (e.g., Ewing's sarcoma, childhood rhabdomyosarcoma, uterine leiomyosarcoma, chondrosarcoma, pulmonary sarcoma, osteosarcoma, and congenital fibrosarcoma), Kaposi's sarcoma, and the like.

[0099] Furthermore, examples of blood cancers include multiple myeloma, malignant lymphomas (e.g., non-Hodgkin's lymphomas (e.g., B-cell non-Hodgkin's lymphomas (e.g., precursor B-cell lymphoblastic lymphoma, precursor B-cell acute lymphoblastic leukemia, chronic B-lymphocytic leukemia, B-cell precursor leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, nodal marginal zone B-cell lymphoma, extranodal marginal zone B-cell lymphoma (MALT lymphoma), primary splenic marginal zone B-cell lymphoma, hairy cell leukemia, hairy cell leukemia-variant type, follicular lymphoma, pediatric follicular lymphoma, diffuse large intestine lymphoma, etc.) B-cell lymphoma, diffuse large B-cell lymphoma, unspecified type, splenic diffuse red pulp small B-cell lymphoma, primary mediastinal large B-cell lymphoma, primary effusion lymphoma, Burkitt lymphoma, mantle cell lymphoma, monoclonal B-cell lymphocytosis, splenic B-cell lymphoma / leukemia, unclassifiable type, monoclonal gammopathy of undetermined significance, IgM type, μ heavy chain disease, λ heavy chain disease, α heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone, extraskeletal plasmacytoma, monoclonal immunoglobulin deposition disease, large B-cell lymphoma with IRF4 rearrangement, primary Cutaneous follicle center lymphoma, T-cell / histiocyte-rich large B-cell lymphoma, primary central nervous system diffuse large B-cell lymphoma, primary cutaneous diffuse large B-cell lymphoma - leg type, EBV-positive diffuse large B-cell lymphoma - not otherwise specified, EBV-positive mucocutaneous ulcer, chronic inflammation-associated diffuse large B-cell lymphoma, lymphomatoid granulomatosis, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, HHV8-positive diffuse large B-cell lymphoma - not otherwise specified, Burkitt-like lymphoma with 11q aberration, MYC and B High-grade B-cell lymphoma with rearrangements of CL2 and / or BCL6, high-grade B-cell lymphoma, unspecified type, and B-cell lymphoma, unclassified type, with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma), T / NK-cell non-Hodgkin lymphoma (e.g., precursor T-cell lymphoblastic lymphoma, chronic T-lymphocytic leukemia, T-cell acute lymphoblastic leukemia, T-cell large granular lymphocytic leukemia, large granular NK-cell leukemia, aggressive NK-cell leukemia, peripheral T-cell lymphoma, peripheral T-cell lymphoma, unspecified type, peripheral T-cell lymphoma, unclassified type),Angioimmunoblastic T-cell lymphoma, anaplastic large cell (CD30 positive) lymphoma, angiocentric lymphoma, intestinal T-cell lymphoma, enteropathy-type T-cell lymphoma, hepatosplenic gamma-delta T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, mycosis fungoides, Sézary syndrome, Hodgkin-like / Hodgkin-related anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma, adult T-cell lymphoma, T-cell prolymphocytic leukemia, chronic NK-cell lymphoproliferative disorder, pediatric allergy Physical EBV-positive T-cell lymphoma, varicella-like bullosa-like lymphoproliferative disorder, extranodal NK / T-cell lymphoma / nasal type, enteropathy-associated T-cell lymphoma, monomorphic epitheliotropic intestinal T-cell lymphoma, gastrointestinal tract slowly progressive T-cell lymphoproliferative disorder hepatosplenic T-cell lymphoma, primary cutaneous CD30-positive T-cell lymphoproliferative disorder, lymphomatoid papulopathy, primary cutaneous anaplastic large cell lymphoma, cutaneous T-cell lymphoma, primary cutaneous γδ T-cell lymphoma, primary cutaneous CD8 Aggressive epidermotropic cytotoxic T-cell lymphoma (ALK-positive), primary cutaneous acral CD8-positive T-cell lymphoma, primary cutaneous CD4-positive small / medium T-cell lymphoproliferative disorder (SMD), follicular T-cell lymphoma, nodal peripheral T-cell lymphoma with follicular helper T-cell phenotype, anaplastic large cell lymphoma (ALK-positive), anaplastic large cell lymphoma (ALK-negative), and breast implant-associated anaplastic large cell lymphoma (ALK-negative) and Hodgkin lymphoma (e.g., classical Hodgkin lymphoma) lymphomas (e.g., nodular sclerosing, mixed cell, lymphocyte-rich, and lymphopenic) and nodular lymphocyte-predominant Hodgkin's lymphoma), leukemias (e.g., acute myeloid leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia (lymphoblastic lymphoma), chronic lymphocytic leukemia (small lymphocytic lymphoma), myelodysplastic syndromes, and chronic myeloid leukemia), primary central nervous system malignant lymphoma, and myeloproliferative syndromes.

[0100] Furthermore, cancers to which the anti-Allergin-1 bispecific antibodies of the present invention are applied also include childhood cancers and cancers of unknown primary origin.

[0101] Furthermore, the anti-Allergin-1 bispecific antibody of the present invention is useful for (a) cancer patients for whom the therapeutic effect of other anti-cancer agents is insufficient or insufficient or whose cancer has progressed after treatment with other anti-cancer agents, (b) patients with curable or inoperable, metastatic, recurrent, refractory and / or distant metastatic cancer, (c) cancer patients in whom the proportion of tumor cells expressing PD-L1 (hereinafter abbreviated as "TPS") or the number of PD-L1 positive cells (tumor cells, lymphocytes and macrophages) divided by the total number of tumor cells multiplied by 100 (hereinafter abbreviated as "CPS") is 50% or more, 25% or more, 10% or more, 5% or more, or 1% or more, (d) cancer patients with MSI-H or dMMR, (e) cancer patients with BRAF (f) patients with malignant melanoma or non-small cell lung cancer that is V600E mutation-positive, (g) patients with cancer that is EGFR gene mutation-positive or ALK fusion gene-positive, or (h) patients with cancer with a high tumor mutation burden (hereinafter abbreviated as "TMB") (10 ６ It may also be applied to patients with cancers with more than 10 mutations per base.

[0102] Here, "other antineoplastic agents" include the antineoplastic agents of the present invention, i.e., alkylating agents, platinum compounds, antimetabolites (e.g., folate antagonists, pyridine metabolism inhibitors, and purine metabolism inhibitors), ribonucleotide reductase inhibitors, nucleotide analogs, topoisomerase inhibitors, microtubule polymerization inhibitors, microtubule depolymerization inhibitors, antitumor antibiotics, cytokine preparations, antihormones, molecularly targeted drugs, tumor immunosuppressants, and other antibody drugs. Furthermore, "the therapeutic effect of other antineoplastic agents is insufficient or not sufficient" refers, for example, to cases where treatment with existing anticancer agents results in "stable disease (SD)" or "progressed disease (PD)" according to RECIST, a tumor shrinkage effect assessment tool.

[0103] Furthermore, the anti-Allergin-1 bispecific antibody of the present invention is useful for (h) cancer patients who have not received treatment with other anti-cancer agents, (i) cancer patients with TPS or CPS of less than 50%, less than 25%, less than 10%, less than 5%, or less than 1%, (j) cancer patients with MSI-H and / or dMMR absent or with MSI-L, (k) patients with BRAF V600 wild-type melanoma or non-small cell lung cancer, (l) patients with EGFR gene mutation-negative and / or ALK fusion gene-negative non-small cell lung cancer, or (m) patients with low TMB (10 ６ It may also be applicable to patients with cancers with fewer than 10 mutations per base.

[0104] Furthermore, the anti-Allergin-1 bispecific antibodies of the present invention can also be applied to postoperative adjuvant therapy to prevent recurrence or metastasis after surgical resection of cancer, or to neoadjuvant therapy performed before surgical resection.

[0105] As used herein, "cancer treatment" includes, for example, treatments performed to (a) reduce cancer cell proliferation, (b) reduce symptoms caused by cancer, (c) improve the quality of life of cancer patients, (d) reduce the dose of other anticancer drugs or adjuvant cancer treatment drugs already administered, and / or (e) extend the survival time of cancer patients. Furthermore, "suppressing cancer progression" means delaying cancer progression, stabilizing cancer-related symptoms, and reversing the progression of symptoms. "Suppressing recurrence" means preventively preventing cancer recurrence in patients whose cancer lesions have completely or substantially disappeared or been removed by cancer treatment or surgical resection.

[0106] The anti-Allergin-1 bispecific antibodies of the present invention are typically administered systemically or locally parenterally. Specific examples of such administration methods include injection, nasal administration, pulmonary administration, and transdermal administration. Examples of injection include intravenous, intramuscular, and intraperitoneal injection. Intravenous administration is preferably by infusion. The dosage varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., but typically ranges from 0.1 μg / kg to 300 mg / kg per adult, and more preferably ranges from 0.1 mg / kg to 10 mg / kg per administration. The dosage is administered parenterally once or several times daily, or by continuous intravenous administration for 30 minutes to 24 hours per day. As mentioned above, the dosage varies depending on various conditions. Therefore, a smaller dosage than the above may be sufficient, or a larger dosage may be required.

[0107] [Combined Uses and Combination Drugs] The anti-Allergin-1 bispecific antibodies of the present invention may be formulated in combination with one or more anti-cancer agents, i.e., as an adjuvant, for the purposes of (a) inhibiting cancer progression, inhibiting recurrence, and / or enhancing the therapeutic effect, (b) reducing the dose of the anti-cancer agent administered in combination, (c) reducing the side effects of the anti-cancer agent administered in combination, and / or (d) enhancing the immunopotentiating effect of the anti-cancer agent administered in combination. In the present invention, when formulated in combination with an anti-cancer agent, the administration form may be a combination drug in which both components are formulated in a single formulation, or may be administered as separate formulations. Such combination use can complement the inhibitory effect of the anti-cancer agent on cancer progression, inhibiting recurrence, and / or enhancing the therapeutic effect, or maintain or reduce the dose or frequency of administration. When the anti-Allergin-1 bispecific antibodies of the present invention and the anti-cancer agent are formulated separately, they may be co-administered for a certain period of time, followed by administration of either the anti-Allergin-1 bispecific antibodies of the present invention or the anti-cancer agent alone. Furthermore, the anti-Allergin-1 bispecific antibody of the present invention may be administered first, followed by the anti-cancer agent, or the anti-cancer agent may be administered first, followed by the antibody of the present invention. Furthermore, the above administration may include a period in which both agents are administered simultaneously. The administration methods for both agents may be the same or different. Depending on the properties of the agents, they may also be provided as a kit of formulations containing the antibody of the present invention and the anti-cancer agent. The dosage of the anti-cancer agent can be appropriately selected based on the dose used in clinical practice. Furthermore, any two or more anti-cancer agents may be administered in combination at an appropriate ratio. Anti-cancer agents include not only those discovered to date but also those to be discovered in the future.

[0108] Other antineoplastic agents include, for example, alkylating agents (e.g., Dacarbazine, Nimustine, Temozolomide, Fotemustine, Bendamustine, Cyclophosphamide, Ifosfamide, Carmustine, Chlorambucil, and Procarbazine, etc.), platinum compounds (e.g., Cisplatin, Carboplatin, Nedaplatin, and Oxaliplatin, etc.), antimetabolites (e.g., antifolates (e.g., Pemetrexed, Leucovorin, and Methotrexate, etc.)), pyridine metabolism inhibitors (e.g., TS-1 (registered trademark), 5-fluorouracil, UFT, Carmofur, and Doxifli, etc.), uridine, FdUrd, Cytarabine, and Capecitabine, etc.), purine metabolism inhibitors (e.g., Fludarabine, Cladribine, and Nelarabine, etc.), ribonucleotide reductase inhibitors, nucleotide analogs (e.g., Gemcitabine, etc.), topoisomerase inhibitors (e.g., Irinotecan, Nogitecan, and Etoposide, etc.), microtubule polymerization inhibitors (e.g., Vinblastine, Vincristine, Vindesine, Vinorelbine, and Eribulin, etc.), microtubule depolymerization inhibitors (e.g., Docetaxel and Paclitaxel, etc.), antitumor antibiotics (e.g., Bleomycin, Mitomycin, etc.), C, Doxorubicin, Daunorubicin, Idarubicin, Etoposide, Mitoxantrone, Vinblastine, Vincristine, Peplomycin, Amrubicin, Aclarubicin, and Epirubicin, etc.), cytokine preparations (e.g., IFN-α2a, IFN-α2b, PEG-IFN-α2b, natural IFN-β, and Interleukin-2, etc.), antihormonal drugs (e.g., Tamoxifen, Fulvestrant, Goserelin, Leuprorelin, Anastrozole, Letrozole, and Exemestane, etc.), molecularly targeted drugs, tumor immunotherapy drugs, and other antibody drugs.Here, the anti-malignant tumor agent according to the present invention is preferably one that is free from the risk of causing cytokine release syndrome when administered alone.

[0109] Examples of molecular targeted drugs include ALK inhibitors (e.g., Crizotinib, Ceritinib, Ensartinib, Alectinib, and Lorlatinib), BCR-ABL inhibitors (e.g., Imatinib and Dasatinib), and EGFR inhibitors (e.g., Erlotinib, EGF816, Afatinib, and Osimertinib). mesilate, Gefitinib, and Rociletinib, etc.), B-RAF inhibitors (e.g., Sorafenib, Vemurafenib, TAK-580, Dabrafenib, Encorafenib, LXH254, Emurafenib, and Zanubrutinib, etc.), VEGFR inhibitors (e.g., Bevacizumab, Apatinib, Lenvatinib, Aflibercept, and Axitinib, etc.), FGFR inhibitors (e.g., AZD4547, Vofatmab, Roblitinib, and Pemigatinib, etc.), c-MET inhibitors (e.g., Savolitinib, Merestinib, Capmatinib, Capmatinib, and Glesatinib, etc.), AXL inhibitors agents (e.g., ONO-7475 and Bemcentinib, etc.), MEK inhibitors (e.g., Cobimetinib, Binimetinib, Selumetinib, and Trametinib, etc.), CDK inhibitors (e.g., Dinaciclib, Abemaciclib, Palbociclib, and Trilaciclib, etc.), BTK inhibitors (e.g., Ibrutinib and Acalabrutinib, etc.), BCL-2 inhibitors (e.g., Navitoclax and Venetoclax, etc.), PI3K-δ / γ inhibitors (e.g., Umbralisib, Parsaclisib, and IPI-549, etc.), JAK-1 / 2 inhibitors (e.g., Itacitinib and Ruxolitinib, etc.), ERK inhibitors (e.g., SCH 900353, etc.), TGFbR1 inhibitors (e.g., Galunisertib, etc.), Cancer cell stemness kinase inhibitors (e.g., Amcasertib), FAK inhibitors (e.g., Defactinib), SYK / FLT3Dual inhibitors (e.g., Mivavotinib, etc.), ATR inhibitors (e.g., Ceralasertib, etc.), WEE1 kinase inhibitors (e.g., Adavosertib, etc.), multi-tyrosine kinase inhibitors (e.g., Sunitinib, Pazopanib, Cabozantinib, Regorafenib, Nintedanib, Sitravatinib, and Midostaurin, etc.), mTOR inhibitors (e.g., Temsirolimus, Everolimus, Vistusertib, and Irinotecan, etc.), HDAC inhibitors (e.g., Vorinostat, Romidepsin, Entinostat, Chidamide, Mocetinostat, Citari nostat, Panobinostat, and Valproate, etc.), PARP inhibitors (e.g., Niraparib, Olaparib, Veliparib, Rucaparib, and Beigene-290, etc.), aromatase inhibitors (e.g., Exemestane and Letrozole, etc.), EZH2 inhibitors (e.g., Tazemetostat, etc.), Galectin-3 inhibitors (e.g., Belapectin, etc.), STAT3 inhibitors (e.g., Napabucasin, etc.), DNMT inhibitors (e.g., Azacitidine, etc.), SMO inhibitors (e.g., Vismodegib, etc.), HSP90 inhibitors (e.g., XL888, etc.), γ-tubulin-specific inhibitors (e.g., Glaziovianin, A and Plinabulin, etc.), HIF2α inhibitors (e.g., PT2385, etc.), glutaminase inhibitors (e.g., telaglenastat, etc.), E3 ligase inhibitors (e.g., avadomide, etc.), NRF2 activators (e.g., omaveloxolone, etc.), arginase inhibitors (e.g., CB-1158, etc.), cell cycle inhibitors (e.g., trabectedin, etc.), Ephrin B4 inhibitors (e.g., sEphB4-HAS, etc.), IAP antagonists (e.g., birinapant, etc.), anti-HER2 antibodies (e.g., pertuzumab, margetuximab, disitamab, disitamab,vedotin, Gancotamab, Timigutuzumab, Zanidatamab, Zenocutuzumab, Trastuzumab, Trastuzumab beta, Trastuzumab deruxtecan, Trastuzumab duocarmazine, Trastuzumab emtansine, R48 and ZW33), anti-HER1 antibody (eg, Cetuximab, Cetuximab sarotalocan, Panitumumab, Necitumumab, Nimotuzumab, Depatuxumab, etc.) mafodotin, futuximab, laprituximab, laprituximab emtansine, matuzumab, modotuximab, petosemtamab, tomuzotuximab, losatuxizumab, losatuxizumab vedotin, serclutamab, serclutamab talirine, imgatuzumab and zalutumumab, etc.), anti-HER3 antibody (eg, Duligotuzumab, Elgemtumab, Istiratumab, Lumretuzumab, Zenocutuzumab, Patritumab, Patritumab, etc.) deruxtecan and Seribantumab, etc.), anti-CD40 antibody (eg, Bleselumab, Dacetuzumab, Iscalimab, Lucatumumab, Mitazalimab, Ravagalimab, Selicrelumab, Teneliximab, ABBV-428 and APX005M), anti-CD70 antibody (eg, Cusatuzumab, Vorsetuzumab, Vorsetuzumab mafodotin and ARGX-110, etc.), anti-VEGF antibody (eg, Bevacizumab, Bevacizumab beta, Ranibizumab, Abiciparpegol, Aflibercept, Brolucizumab, Conbercept, Dilpacimab, Faricimab, Navixixizumab, Varisacumab, and IMC-1C11, etc.), anti-VEGFR1 antibodies (e.g., Icrucumab, etc.), anti-VEGFR2 antibodies (e.g., Ramucirumab, Alacizumab, Alacizumab pegol, Olinvacimab, Pegdinetanib, and AMG596, etc.), anti-CD20 antibodies (e.g., Rituximab, Blontuvetmab, Epitumomab, Ibritumomab tiuxetan, Ocaratuzumab, Ocrelizumab, Technetium ( ９９ mTc) nofetumomab merpentan, Tositumomab, Veltuzumab, Ofatumumab, Ublituximab, Obinutuzumab, and Nofetumomab, etc.), anti-CD30 antibodies (e.g., Brentuximab Vedotin and Iratumumab, etc.), anti-CD38 antibodies (e.g., Daratumumab, Isatuximab, Mezagitamab, AT13 / 5, and MOR202, etc.), anti-TNFRSF10B antibodies (e.g., Benufutamab, Conatumumab, Drozitumab, Lexatumumab, Tigatuzumab, Eftozanermin alfa, and DS-8273a, etc.), anti-TNFRSF10A antibodies (e.g., Mapatumumab, etc.), anti-MUC1 antibodies (e.g., Cantuzumab, Cantuzumab ravtansine, Crivatuzumab, Crivatuzumab tetraxetan, Yttrium ( ９０Y) clivatuzumab tetraxetan, Epitumomab, Epitumomab cituxetan, Sontuzumab, Gatipotuzumab, Nacolomab, Nacolomab tafenatox, 7F11C7, BrE-3, CMB-401, CTM01, and HMFG1, etc.), anti-MUC5AC antibodies (e.g., Ensituximab, etc.), anti-MUC16 antibodies (e.g., Oregovomab, Abagovomab, Igovomab, and Sofituzumab vedotin, etc.), anti-DLL4 antibodies (e.g., Demcizumab, Dilpacimab, Navicizumab, and Enoticumab, etc.), anti-fucosyl-GM1 antibodies (e.g., BMS-986012, etc.), anti-gpNMB antibodies (e.g., Glembatumumab vedotin, etc.), anti-Mesothelin antibodies (e.g., Amatuximab, Anetumab ravtansine, Anetumab corixetan, RG7784, and BMS-986148, etc.), anti-MMP9 antibodies (e.g., Andecaliximab, etc.), anti-GD2 antibodies (e.g., Dinutuximab, Dinutuximab beta, Lorukafusp alfa, Naxitamab, 14G2a, MORAb-028, Surek, TRBs07, and ME361, etc.), anti-MET antibodies (e.g., Emibetuzumab, Onartuzumab, Telisotuzumab, and Telisotuzumab vedotin, etc.), anti-FOLR1 antibodies (e.g., Farletuzumab, Mirvetuximab, and Mirvetuximab soravtansine, etc.), anti-CD79b antibodies (e.g., Iladatuzumab, Iladatuzumab vedotin, and Polatuzumab vedotin, etc.), anti-DLL3 antibodies (e.g., Rovalpituzumab and Rovalpituzumab Tesirine, etc.), anti-CD51 antibodies (e.g., Abituzumab, Etaracizumab, and Intetumumab, etc.), anti-EPCAM antibodies (e.g., Adecatumumab, Catumaxomab, Edrecolomab, Oportuzumab, etc.),monatox, Citatuzumab bogatox and Tucotuzumab celmoleukin), anti-CEACAM5 antibodies (e.g. Altumomab, Arcitumomab, Cergutuzumab amunaleukin, Labetuzumab, Labetuzumab govitecan, ９０ Y-cT84.66, AMG211, BW431 / 26, CE25 / B7, COL-1 and T84.66 M5A, etc.), anti-CEACAM6 antibodies (e.g., Tinurilimab, etc.), anti-FGFR2 antibodies (e.g., Aprutumab, Aprutumab ixadotin and Bemarituzumab, etc.), anti-CD44 antibodies (e.g., Bivatuzumab mertansine, etc.), anti-PSMA antibodies (e.g., Indium ( １１１ In) capromab pendetide, １７７ Lu-J591 and ES414, etc.), anti-Endoglin antibodies (e.g., Carotuximab, etc.), anti-IGF1R antibodies (e.g., Cixutumumab, Figitumumab, Ganitumab, Dalotuzumab, Teprotumumab, and Robatumumab, etc.), anti-TNFSF11 antibodies (e.g., Denosumab), anti-GUCY2C antibodies (e.g., Indusatumab vedotin, etc.), anti-SLC39A6 antibodies (e.g., Ladiratuzumab vedotin, etc.), anti-SLC34A2 antibodies (e.g., Lifastuzumab vedotin, etc.), anti-NCAM1 antibodies (e.g., Lorvotuzumab mertansine and N901, etc.), anti-ganglioside GD3 antibodies (e.g., Ecromeximab and Mitumomab, etc.), anti-AMHR2 antibodies (e.g., Murlentamab, etc.), anti-CD37 antibodies (e.g., Lilotomab, Lutetium ( １７７lu) lilotomab satetraxistan, Naratuximab, Naratuximab emtansine, and Otlertuzumab, etc.), anti-IL1RAP antibodies (e.g., Nidanilimab, etc.), anti-PDGFR2 antibodies (e.g., Olaratumab and Tovetumab, etc.), anti-CD200 antibodies (e.g., Samalizumab, etc.), anti-TAG-72 antibodies (e.g., Anatumomab mafenatox, Minretumomab, Indium ( １１１ In) satumomab pendetide, CC49, HCC49, and M4, etc.), anti-SLITRK6 antibodies (e.g., sirtratumab vedotin, etc.), anti-DPEP3 antibodies (e.g., tamrintamab pamozirine, etc.), anti-CD19 antibodies (e.g., axicabtagene ciloleucel, coltuximab ravtansine, denintuzumab mafodotin, inebilizumab, loncastuximab, loncastuximab tesirine, obexelimab, tafasitamab, taplitumomab paptox, and huAnti-B4, etc.), anti-NOTCH2 / 3 antibodies (e.g., tarextumab, etc.), anti-tenascin C antibodies (e.g., tenatumomab, etc.), anti-AXL antibodies (e.g., enapotamab, enapotamab vedotin, tilvestamab, etc.), anti-STEAP1 antibodies (e.g., vandortuzumab vedotin, etc.), anti-CTAA16 antibodies (e.g., technetium ( ９９mTc) votumumab, etc.), anti-CLDN18 antibodies (e.g., Zolbetuximab, etc.), anti-GM3 antibodies (e.g., Racotumomab, FCGR1, and H22, etc.), anti-PSCA antibodies (e.g., MK-4721, etc.), anti-FN extra domain B antibodies (e.g., AS1409, etc.), anti-HAVCR1 antibodies (e.g., CDX-014, etc.), anti-TNFRSF4 antibodies (e.g., MEDI6383, etc.), anti-HER1-MET bispecific antibodies (e.g., Amivantamab), anti-EPCAM-CD3 bispecific antibodies (e.g., Solitomab and Catumaxomab, etc.), anti-Ang2-VEGF bispecific antibodies (e.g., Vanucizumab, etc.), anti-HER2-CD3 bispecific antibodies (e.g., Ertumaxom ab, etc.), anti-HER3-IGF1R bispecific antibodies (e.g., Istiratumab, etc.), anti-PMSA-CD3 bispecific antibodies (e.g., Pasotuxizumab, etc.), anti-HER1-LGR5 bispecific antibodies (e.g., Petosemtamab, etc.), anti-SSTR2-CD3 bispecific antibodies (e.g., Tidutamab, etc.), anti-CD30-CD16A bispecific antibodies (e.g., AFM13, etc.), anti-CEA-CD3 bispecific antibodies (e.g., Cibisatamb and RO6958688), anti-CD3-CD19 bispecific antibodies (e.g., Duvortuxizumab and Blinatumomab, etc.), anti-IL3RA-CD3 bispecific antibodies (e.g., Flotetuzumab and Vibecotamab, etc.), anti-GPRC5D-CD3 bispecific antibodies (e.g., Talquetamab, etc.), anti-CD20-CD3 bispecific antibodies (e.g., Plamotamab, Odronextamab, Mosunetuzumab, Glof itamab, Epcoritamab, and REGN1979, etc.), anti-TNFRSF17-CD3 bispecific antibodies (e.g., Teclistamab, etc.), anti-CLEC12A-CD3 bispecific antibodies (e.g., Tepoditamab, etc.), anti-HER2-HER3 bispecific antibodies (e.g., Zenocutuzumab, etc.), anti-FAP antibody / IL-2 fusion proteins (e.g., RO6874281, etc.), and anti-CEA antibody / IL-2 fusion proteins (e.g., Cergutuzumab, etc.),amunaleukin, etc.).

[0110] Furthermore, examples of tumor immunotherapeutic agents include anti-PD-1 antibodies (e.g., Nivolumab, Cemiplimab, Pembrolizumab, Spartalizumab, Tislelizumab, Dostarlimab, Toripalimab, Camrelizumab, Genolimzumab, Sintilimab, Lodapolimab, Retifanlimab, Balstilimab, Serplulimab, Budigalimab, Prolgolimab, Sasanlimab, Cetrelimab, Zimberelimab, Geptanolimab, AMP-514, STI-A1110, ENUM 388D4, ENUM 244C8, Zimberelimab, CS1003, BAT-1306, Penpulimab, AK103, Ezabenlimab, LZM009, CMAB819, Sym021, SSI-361, JY034, Pucotenlimab, ISU106, and CX-188, etc.), anti-PD-L1 antibodies (e.g., Atezolizumab, Avelumab, Durvalumab, Manelimab, Pacmilimab, Envafolimab, Cosibelimab, Sugemalimab, BMS-936559, STI-1014, HLX20, Adebrelimab, MSB2311, BGB-A333, KL-A167, AK106, Socazolimab, FAZ053, CBT-502, and and JS003, etc.), PD-1 antagonists (e.g., AUNP-12, the compounds BMS-M1 to BMS-M10 (WO2014 / 151634, WO2016 / 039749, WO2016 / 057624, WO2016 / 077518, WO2016 / 100285, WO2016 / 100608, WO2016 / 126646, WO2016 / 149351, WO201 7 / 151830 and WO2017 / 176608), BMS-1, BMS-2, BMS-3, BMS-8, BMS-37, BMS-200, BMS-202, BMS-230, BMS-242, BMS-1001, and BMS-1166 (see WO2015 / 034820, WO2015 / 160641, WO2017 / 066227, and Oncotarget. 2017 Sep 22;8(42):72167-72181), Incyte-1 to Incyte-6 compounds (see WO2017 / 070089, WO2017 / 087777, WO2017 / 106634, WO2017 / 112730, WO2017 / 192961 and WO2017 / 205464), CAMC-1 to CAMC-4 (see WO2017 / 202273, WO2017 / 202274, WO2017 / 202275 and WO2017 / 202276), RG_1 (see WO2017 / 118762), and DPPA-1 (Angew. Chem. Int. Ed. 2015, 54,11760-11764), PD-L1 / VISTA antagonists (e.g., CA-170), PD-L1 / TIM3 antagonists (e.g., CA-327), anti-PD-L2 antibodies, PD-L1 fusion proteins, PD-L2 fusion proteins (e.g., AMP-224), anti-CTLA-4 antibodies (e.g., Ipilimumab, Zalifrelimab, Nurulimab, Tremelimumab, etc.), anti-LAG-3 antibodies (e.g., Relatlimab, Ieramilimab, Fianlimab, Encelimab, Mavezelimab, etc.), anti-TIM3 antibodies (e.g., MBG453, Cobolimab, etc.), anti-KIR antibodies (e.g., Lirilumab, IPH2101, LY3321367, MK-4280, etc.), anti-BTLA antibodies, anti-TIGIT antibodies (e.g., Tiragolumab, Etigilimab, etc.), , Vibostolimab, and BMS-986207, etc.), anti-VISTA antibodies (e.g., Onvatilimab, etc.), anti-CD137 antibodies (e.g., Urelumab and Utomilumab, etc.), anti-CSF-1R antibodies / CSF-1R inhibitors (e.g., Cabiralizumab, Emactuzumab, LY3022855, Axatilimab, MCS-110, IMC-CS4, AMG820, Pexid artinib, BLZ945, and ARRY-382, etc.), anti-OX40 antibodies (e.g., MEDI6469, Ivuxolimab, MEDI0562, MEDI6383, Efizonerimod, GSK3174998, BMS-986178, and MOXR0916, etc.), anti-HVEM antibodies, anti-CD27 antibodies (e.g., Varlilumab, etc.), anti-GITR antibodies / GITR fusion proteins (e.g., Efaprinermin alfa, Efgivanermin alfa, MK-4166, INCAGN01876, GWN323, and TRX-518, etc.), anti-CD28 antibodies, anti-CCR4 antibodies (e.g., Mogamulizumab, etc.), anti-B7-H3 antibodies (e.g., Enoblituzumab, Mirzotamab, Mirzotamab, etc.),clezutoclax and Omburtamab, etc.), anti-ICOS agonist antibodies (e.g., Vopratelimab and GSK3359609, etc.), anti-CD4 antibodies (e.g., Zanolimumab and IT1208, etc.), anti-DEC-205 antibody / NY-ESO-1 fusion protein (e.g., CDX-1401, etc.), anti-SLAMF7 antibodies (e.g., Azintuxizumab, vedotin and elotuzumab, etc.), anti-CD73 antibodies (e.g., oleclumab and BMS-986179, etc.), pegylated IL-2 (bempegaldesleukin), IDO inhibitors (e.g., epacadostat, indoximod, linrodostat, etc.), TLR agonists (e.g., motolimod, CMP-001, G100, tilsotolimod, SD-101, and MEDI9197, etc.), adenosine A2A receptor antagonists (e.g., preladenant, AZD4635, taminadenant, ciforadenant, etc.), anti-NKG2A antibodies (e.g., monalizumab, etc.), anti-CSF-1 antibodies (e.g., PD0360324, etc.), immune enhancers (e.g., PV-10, etc.), IL-15 superagonists (e.g., ALT-803, etc.), soluble LAG3 (e.g., eftilagimod, alpha, etc.), anti-CD47 antibodies / CD47 antagonists (e.g., ALX148, etc.), and IL-12 antagonists (e.g., M9241, etc.). Nivolumab can be produced according to the method described in WO2006 / 121168, Pembrolizumab can be produced according to the method described in WO2008 / 156712, BMS-936559 can be produced according to the method described in WO2007 / 005874, and Ipilimumab can be produced according to the method described in WO2001 / 014424.

[0111] Further examples of other antibody pharmaceuticals include anti-IL-1β antibodies (e.g., canakinumab) and anti-CCR2 antibodies (e.g., plozalizumab). When used in combination with the anti-Allergin-1 bispecific antibody of the present invention, the tumor immunotherapeutic drugs of the present invention can be administered, for example, according to the following dosage and administration method: The active ingredient of each tumor immunotherapeutic drug is administered intravenously (e.g., by intravenous drip infusion) at a dose of about 1 to about 21 mg / kg (body weight) or about 80 to about 1680 mg per dose over a period of about 30 to about 60 minutes or over about 60 minutes or longer, at intervals of 1 to 8 weeks. Here, examples of the dose per administration in terms of body weight include 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 12 mg / kg, 14 mg / kg, 15 mg / kg, 20 mg / kg, and 21 mg / kg, while examples of the dose per administration include 200 mg, 240 mg, 250 mg, 280 mg, 300 mg, 320 mg, 350 mg, 360 mg, 400 mg, 420 mg, 450 mg, 480 mg, 500 mg, 540 mg, 560 mg, 600 mg, 640 mg, 700 mg, 720 mg, 750 mg, 800 mg, 840 mg, 900 mg, 960 mg, 1000 mg, 1080 mg, and the like. The dosage ranges include 1, 2, 3, 4, 6, and 8 weeks, and the duration of a single administration is, for example, about 30 minutes, about 60 minutes, or more than about 60 minutes.

[0112] Here, when the active ingredient of the tumor immunotherapeutic agent is the anti-PD-1 antibody nivolumab, and when it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, nivolumab can be administered to adults by intravenous infusion at (a) 1 mg / kg (body weight) every 3 weeks, (b) 3 mg / kg (body weight) every 2 weeks, (c) 2 mg / kg (body weight) every 3 weeks, (d) 80 mg every 3 weeks, (e) 240 mg every 2 weeks, (f) 360 mg every 3 weeks, or (g) 480 mg every 4 weeks (preferably 240 mg every 2 weeks, 360 mg every 3 weeks, or 480 mg every 4 weeks). When the active ingredient of the tumor immunotherapy drug is the anti-PD-1 antibody pembrolizumab, and when it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, pembrolizumab may be administered to adults by intravenous infusion at (a) 200 mg once every 3 weeks, (b) 400 mg once every 6 weeks, or (c) 2 mg / kg (body weight) once (up to a maximum of 200 mg per administration) once every 3 weeks.

[0113] When the active ingredient of the tumor immunotherapy drug is the anti-PD-1 antibody cemiplimab-rwlc, and when it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, cemiplimab-rwlc can be administered intravenously to adults at a dose of 350 mg every three weeks. In particular, it can be administered to patients with squamous cell carcinoma in the same dosage regimen as described above.

[0114] When the active ingredient of the tumor immunotherapy drug is the anti-PD-1 antibody Dostarlimab, and it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, the drug can be administered to adults by intravenous infusion at a dose of 500 mg every three weeks for four doses, followed by intravenous infusion at a dose of 1000 mg every six weeks from the fifth dose three weeks later. In particular, the drug can be administered in the same dosage regimen as above for dMMR-positive endometrial cancer or solid tumors.

[0115] When the active ingredient of the tumor immunotherapy drug is the anti-PD-L1 antibody Avelumab, and it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, it can be administered to adults by intravenous infusion at a dose of 10 mg / kg (body weight) of Avelumab once every two weeks.

[0116] When the active ingredient of the tumor immunotherapy drug is the anti-PD-L1 antibody atezolizumab, and it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, atezolizumab may be administered to adults by intravenous infusion at (a) 840 mg once every 2 weeks, (b) 1200 mg once every 3 weeks, or (c) 1680 mg once every 4 weeks.

[0117] When the active ingredient of the tumor immunotherapy drug is the PD-L1 antibody durvalumab, and it is used in combination with the anti-Allergin-1 bispecific antibody of the present invention, durvalumab can be administered to adults by intravenous infusion at a dose of 10 mg / kg (body weight) once every two weeks.

[0118] When the active ingredient of the tumor immunotherapy drug is the anti-CTLA-4 antibody ipilimumab, when used in combination with the anti-Allergin-1 bispecific antibody of the present invention, ipilimumab can be administered to adults by intravenous infusion at (a) 3 mg / kg (body weight) once daily, or (b) 1 mg / kg (body weight) once daily, four times at three-week intervals.

[0119] [Formulations] When the anti-Allergin-1 bispecific antibodies of the present invention are formulated and used as injections or infusions for intravenous drip infusion, the injections or infusions may be in the form of an aqueous solution, suspension, or emulsion, or may be formulated as a solid together with a pharmaceutically acceptable carrier so that they can be dissolved, suspended, or emulsified by adding a solvent when used. Solvents that can be used for injections or infusions for intravenous drip infusion include, for example, distilled water for injection, physiological saline, glucose solution, and isotonic solutions (e.g., solutions of sodium chloride, potassium chloride, glycerin, mannitol, sorbitol, boric acid, borax, propylene glycol, etc.).

[0120] Examples of pharmaceutically acceptable carriers include stabilizers, solubilizers, suspending agents, emulsifiers, soothing agents, buffers, preservatives, antiseptics, pH adjusters, and antioxidants. Examples of stabilizers that can be used include various amino acids, albumin, globulin, gelatin, mannitol, glucose, dextran, ethylene glycol, propylene glycol, polyethylene glycol, ascorbic acid, sodium bisulfite, sodium thiosulfate, sodium edetate, sodium citrate, and dibutylhydroxytoluene. Examples of solubilizing agents that can be used include alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants (e.g., Polysorbate 20 (registered trademark), Polysorbate 80 (registered trademark), HCO-50, and the like). Examples of suspending agents that can be used include glycerin monostearate, aluminum monostearate, methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, and sodium lauryl sulfate. Examples of emulsifiers that can be used include gum arabic, sodium alginate, and tragacanth. Examples of soothing agents that can be used include benzyl alcohol, chlorobutanol, and sorbitol. Examples of buffers that can be used include phosphate buffer, acetate buffer, borate buffer, carbonate buffer, citrate buffer, Tris buffer, glutamate buffer, and epsilon aminocaproic acid buffer. Examples of preservatives that can be used include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, propyl parahydroxybenzoate, butyl parahydroxybenzoate, chlorobutanol, benzyl alcohol, benzalkonium chloride, sodium dehydroacetate, sodium edetate, boric acid, and borax. Examples of antiseptics that can be used include benzalkonium chloride, parahydroxybenzoic acid, and chlorobutanol. Examples of pH adjusters that can be used include hydrochloric acid, sodium hydroxide, phosphoric acid, and acetic acid.Examples of antioxidants that can be used include: (1) water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, lecithin, propyl gallate, and α-tocopherol; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid, sorbitol, tartaric acid, and phosphoric acid.

[0121] Injections or infusion solutions for intravenous drip can be produced by sterilizing in the final step or by aseptic procedures, for example, by sterilizing by filtration through a filter or the like, and then filling into sterile containers. Alternatively, injections or infusion solutions for intravenous drip can be prepared by vacuum-drying or lyophilizing sterile powders (which may contain a powder of a pharmaceutically acceptable carrier) and dissolving them in an appropriate solvent before use. The present invention provides, for example, the following embodiments [1] to

[150] , [1-1] and [1-2], [2-1] to [2-32], [3-1] to [3-3], [4-1], [5-1], [6-1], [7-1] to [7-20], and [8-1] to [8-3]. [1] A bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1, wherein the antigen-binding sites do not compete with each other for Allergin-1, and the two different antigen-binding sites are selected from the group consisting of: (i) an antigen-binding site that specifically binds to a specific epitope in the S1 domain of Allergin-1 (S1-A binding site); (ii) an antigen-binding site that specifically binds to another epitope in the S1 domain of Allergin-1 that is different from the epitope described in (i) above (S1-B binding site); (iii) an antigen-binding site that specifically binds to a specific epitope in the S2 domain of Allergin-1 (S2-C binding site); and (iv) an antigen-binding site that specifically binds to another epitope in the S2 domain of Allergin-1 that is different from the epitope described in (iii) above (S2-D binding site), wherein the S1-A binding site is (A) an antigen-binding site consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (B) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (C) an antigen-binding site consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (D) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, wherein the S1-B binding site is any one of the antigen-binding sites selected from the group consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63, or (F) (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64,(e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, wherein the S2-C binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, wherein the S2-D binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72; [2] A bispecific antibody or antibody fragment thereof against Allergin-1, comprising any two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, wherein the S1-A binding site is: (A) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (B) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30; and(d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (C) (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (D) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36; (d) an antigen-binding site selected from the group consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, wherein the S1-B binding site is an antigen-binding site selected from the group consisting of: (E) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63, or (F) an antigen-binding site consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66,the S2-C binding site is an antigen-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69; and the S2-D binding site is an antigen-binding site consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, (f) the bispecific antibody or antibody fragment thereof, wherein the antigen-binding site consists of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72; [3] the bispecific antibody or antibody fragment thereof according to the above item [1] or [2], wherein any 1 to 5 amino acid residues in one or more CDRs of VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2 and VL-CDR3 constituting any two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site and an S2-D binding site may be substituted with other amino acids; [4] the bispecific antibody or antibody fragment thereof, wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, and (i) the S1-A binding site is composed of: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26 and (c) an antigen-binding site consisting of a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51, (ii) an S1-B binding site consisting of: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and an antigen-binding site consisting of a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; [5] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, and (i) the S1-A binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; and (ii) an S1-B-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66; [6] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A-binding site and an S1-B-binding site, and (i) the S1-A-binding site is (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and(f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54, and (ii) an S1-B-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; [7] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A-binding site and an S1-B-binding site, and (i) the S1-A-binding site is consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54, (ii) an S1-B binding site comprising: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66; [8] the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, and (i) the S1-A binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33;(d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (ii) an S1-B-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; [9] a bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A-binding site and an S1-B-binding site, (i) the S1-A binding site is an antigen-binding site consisting of: (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33; and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; (ii) the S1-B binding site is: (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is composed of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66;

[10] the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, and (i) the S1-A binding site comprises: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, (ii) an S1-B binding site comprising: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is composed of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63;

[11] the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, (i) the S1-A binding site is composed of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, and (ii) the S1-B binding site is composed of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, a bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VH having (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66;

[12] The two different antigen-binding sites are an S1-A binding site and an S2-C binding site, (i) the S1-A binding site is an antigen-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51, and (ii) the S2-C binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and

[13] The bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[13] The two different antigen-binding sites are an S1-A binding site and an S2-C binding site, and (i) the S1-A binding site is an antigen-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54, (ii) a VH having an S2-C binding site comprising: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43; (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44; and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67; (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68;(f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is composed of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[14] the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, (i) the S1-A binding site is composed of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57, and (ii) the S2-C binding site is composed of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[15] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, (ii) an S2-C binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and

[16] The bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[16] The two different antigen-binding sites are an S1-A binding site and an S2-D binding site, and (i) the S1-A binding site is an antigen-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (ii) a bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the S2-D-binding site is an antigen-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[17] a bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A-binding site and an S2-D-binding site, and (i) the S1-A-binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) an VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54, (ii) an S2-D binding site consisting of: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[18] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (ii) an S2-D-binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[19] a bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-A-binding site and an S2-D-binding site, and (i) the S1-A-binding site is (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and(f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, and (ii) an S2-D binding site consisting of: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[20] A bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-B binding site and an S2-C binding site, and (i) the S1-B binding site is consisting of: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63, (ii) an S2-C binding site comprising: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[21] the two different antigen-binding sites are an S1-B binding site and an S2-C binding site, and (i) the S1-B binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42;(d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66; and (ii) an S2-C binding site consisting of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69;

[22] a bispecific antibody or an antibody fragment thereof according to the preceding item [1] or [2], wherein the two different antigen-binding sites are an S1-B binding site and an S2-D binding site, (i) the S1-B binding site is an antigen-binding site consisting of: (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39; and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; (ii) the S2-D binding site is: (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71; (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is composed of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[23] the two different antigen-binding sites are an S1-B binding site and an S2-D binding site, and (i) the S1-B binding site comprises: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, (ii) an S2-D binding site comprising: (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) the bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the antigen-binding site is composed of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[24] the two different antigen-binding sites are an S2-C binding site and an S2-D binding site, (i) the S2-C binding site is composed of a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, and (ii) the S2-D binding site is composed of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, a bispecific antibody or antibody fragment thereof according to the preceding item [1] or [2], wherein the bispecific antibody or antibody fragment thereof has an antigen-binding site consisting of: (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72;

[25] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[24] , wherein the FW of the VH constituting each of two different antigen-binding sites selected from the S1-A-binding site, the S1-B-binding site, the S2-C-binding site, and the S2-D-binding site consists of an amino acid sequence encoded by the germline VH gene IGHV3-66 or the gene which has undergone one or more somatic mutations and / or back mutations, and / or the FW of the VL constituting each of the antigen-binding sites consists of an amino acid sequence encoded by the germline VL gene IGKV1-39 or the gene which has undergone one or more somatic mutations and / or back mutations;

[26] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[24] , wherein the S1-A-binding site is selected from the following: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) a pair of amino acid sequences selected from SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) a pair of amino acid sequences selected from SEQ ID NO: 6 and SEQ ID NO: 17 or 109, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to said amino acid sequences (excluding the three CDRs of each);

[27] a bispecific antibody or an antibody fragment thereof according to any one of the preceding items [1] to

[25] , wherein the S1-B binding site is selected from: (a) a pair of amino acid sequences selected from SEQ ID NO: 7 and SEQ ID NO: 18 or 110, or (b) a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[26] , which is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 8 and SEQ ID NO: 19 or 111, or a pair of amino acid sequences which are independently at least 80%, 90%, 95%, 98%, or 99% identical to said amino acid sequences (excluding the three CDRs of each);

[28] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[27] , wherein the S2-C binding site is composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112, or a pair of amino acid sequences which are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs);

[29] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[28] , wherein the S2-D binding site is composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114, or a pair of amino acid sequences which are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs);

[30]

[31] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[25] ,

[27] , and

[30] , wherein the S1-A binding site is composed of a VH and a VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) SEQ ID NO: 6 and SEQ ID NO: 17 or 109;

[32] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[25] ,

[27] , and

[30] , wherein the S1-B binding site is composed of a VH and a VL consisting of a pair of amino acid sequences: (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110, or (b) SEQ ID NO: 8 and SEQ ID NO: 19 or 111;

[32]

[33] a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[25] ,

[28] ,

[30] and

[31] , wherein the S2-C binding site is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112;

[33] a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[25] ,

[28] ,

[30] and

[31] , wherein the S2-C binding site is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112;(b) a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[25] and

[29] to

[32] , which is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[34] a bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1, wherein the antigen-binding sites do not compete with each other for Allergin-1 and are two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, wherein (i) the S1-A binding site is selected from: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) the VH and VL are composed of a pair of amino acid sequences selected from SEQ ID NO: 6 and SEQ ID NO: 17 or 109, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs); (ii) the S1-B binding site is composed of a VH and VL are composed of a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs); (iii) the S2-C binding site is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding each of the three CDRs); and (iv) the S2-D binding site is composed of: (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or(b) a bispecific antibody or antibody fragment thereof, which is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 22 or 114, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding three CDRs);

[35] a bispecific antibody or antibody fragment thereof against Allergin-1, which consists of any two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, wherein (i) the S1-A binding site is selected from: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) the VH and VL are composed of a pair of amino acid sequences selected from SEQ ID NO: 6 and SEQ ID NO: 17 or 109, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs); (ii) the S1-B binding site is composed of a VH and VL are composed of a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding the three CDRs); (iii) the S2-C binding site is composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding each of the three CDRs); and (iv) the S2-D binding site is composed of: (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or(b) a bispecific antibody or antibody fragment thereof, which is composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114, or a pair of amino acid sequences that are at least 80%, 90%, 95%, 98%, or 99% independently identical to each of said amino acid sequences (excluding three CDRs);

[36] (i) the S1-A binding site is composed of a VH and a VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO: 1 and SEQ ID NO: 12 or 104, (b) SEQ ID NO: 2 and SEQ ID NO: 13 or 105, (c) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (d) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (e) SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and (f) SEQ ID NO: 6 and SEQ ID NO: 17 or 109; and (ii) the S1-B binding site is composed of a VH and a VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NO: 7 and SEQ ID NO: 18 or 110, or (b) the bispecific antibody or antibody fragment thereof according to the preceding item

[34] or

[35] , wherein (b) the VH and VL comprise a VH and VL comprising a pair of amino acid sequences set forth in SEQ ID NO: 8 and SEQ ID NO: 19 or 111; (iii) the VH and VL comprising the S2-C binding site comprise a VH and VL comprising ...; a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 12 or 104, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 18 or 110;

[38] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 13 or 105, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 7 and SEQ ID NO: 18 or 110;

[39]

[40] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 14 or 106, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 7 and 18 or 110;

[41] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 15 or 107, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 7 and 18 or 110;

[41] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 18 or 110;

[42] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 7 and SEQ ID NO: 18 or 110;

[43]

[43] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 1 and 12 or 104, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 8 and 19 or 111;

[44] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 2 and 13 or 105, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 8 and 19 or 111;

[45] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 3 and SEQ ID NO: 14 or 106, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 8 and SEQ ID NO: 19 or 111;

[46] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 4 and SEQ ID NO: 15 or 107, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 8 and SEQ ID NO: 19 or 111;

[47]

[48] ​​A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 5 and 16 or 108, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 8 and 19 or 111;

[49] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S1-B binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 6 and 17 or 109, and the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 8 and 19 or 111;

[49] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 12 or 104, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112;

[50] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 13 or 105, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112;

[51]

[51] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 14 or 106, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 9 and 20 or 112;

[52] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 15 or 107, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 9 and 20 or 112;

[53] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112;

[54] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-C binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112;

[55]

[56] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 1 and SEQ ID NO: 12 or 104, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[57] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 13 or 105, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113; a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 3 and SEQ ID NO: 14 or 106, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[58] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 4 and SEQ ID NO: 15 or 107, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[59]

[60] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[61] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113; a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 12 or 104, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[62] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 13 or 105, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[63]

[64] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 3 and SEQ ID NO: 14 or 106, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[65] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 4 and SEQ ID NO: 15 or 107, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114; a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 16 or 108, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[66] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-A binding site and an S2-D binding site, the S1-A binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[67]

[68] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B binding site and an S2-C binding site, the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 7 and 18 or 110, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 9 and 20 or 112;

[69] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B binding site and an S2-C binding site, the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 8 and 19 or 111, and the S2-C binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 9 and 20 or 112;

[69] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B-binding site and an S2-D-binding site, the S1-B-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 7 and SEQ ID NO: 18 or 110, and the S2-D-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[70] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] , and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B-binding site and an S2-D-binding site, the S1-B-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 7 and SEQ ID NO: 18 or 110, and the S2-D-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[71]

[72] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B binding site and an S2-D binding site, the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 8 and SEQ ID NO: 19 or 111, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[73] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S1-B binding site and an S2-D binding site, the S1-B binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 8 and SEQ ID NO: 19 or 111, and the S2-D binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114; a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S2-C-binding site and an S2-D-binding site, the S2-C-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and the S2-D-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[74] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1], [2],

[25] and

[34] to

[36] , wherein the two different antigen-binding sites are an S2-C-binding site and an S2-D-binding site, the S2-C-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and the S2-D-binding site being composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and SEQ ID NO: 22 or 114;

[75] A bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1, wherein the antigen-binding sites do not compete with each other for Allergin-1 and are two different antigen-binding sites selected from an S1-A-binding site, an S1-B-binding site, an S2-C-binding site and an S2-D-binding site, and wherein (i) the S1-A-binding site is: (a) SEQ ID NO: 1 and SEQ ID NO: 104, (b) SEQ ID NO: 2 and SEQ ID NO: 105, (c) SEQ ID NO: 3 and SEQ ID NO: 106, (d) SEQ ID NO: 4 and SEQ ID NO: 107, (e) SEQ ID NO: 5 and SEQ ID NO: 108, and (f) SEQ ID NO: 6 and SEQ ID NO: 109; (ii) the S1-B binding site is capable of cross-competing for binding to the S1 domain of Allergin-1 by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of (a) SEQ ID NO: 7 and SEQ ID NO: 110, or (b) SEQ ID NO: 8 and SEQ ID NO: 111; (iii) the S2-C binding site is capable of cross-competing for binding to the S2 domain of Allergin-1 by an antibody or antigen-binding site thereof having a VH and VL consisting of the pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 112; (iv) the S2-D binding site is (a) a bispecific antibody or an antibody fragment thereof that can cross-compete for binding to the S2 domain of Allergin-1 by an antibody or antigen-binding site thereof having a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 10 and 113, or (b) SEQ ID NOs: 11 and 114;

[76] A bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1, wherein the antigen-binding sites do not compete with each other for Allergin-1 and are any two different antigen-binding sites selected from the group consisting of an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, and (i) the S1-A binding site is an antigen-binding site whose binding to the S1 domain of Allergin-1 is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of any one pair of amino acid sequences selected from the group consisting of: (a) SEQ ID NO: 1 and SEQ ID NO: 104, (b) SEQ ID NO: 2 and SEQ ID NO: 105, (c) SEQ ID NO: 3 and SEQ ID NO: 106, (d) SEQ ID NO: 4 and SEQ ID NO: 107, (e) SEQ ID NO: 5 and SEQ ID NO: 108, and (f) SEQ ID NO: 6 and SEQ ID NO: 109, (ii) the S1-B binding site is an antigen-binding site whose binding to the S1 domain of Allergin-1 is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of a pair of amino acid sequences of SEQ ID NOs: 7 and 110, or (b) SEQ ID NOs: 8 and 111; (iii) the S2-C binding site is an antigen-binding site whose binding to the S2 domain of Allergin-1 is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of a pair of amino acid sequences of SEQ ID NOs: 9 and 112; and (iv) the S2-D binding site is an antigen-binding site whose binding to the S2 domain of Allergin-1 is cross-competed by an antibody or antigen-binding site thereof having a VH and VL consisting of a pair of amino acid sequences of SEQ ID NOs: 10 and 113, or (b) SEQ ID NOs: 11 and 114;

[78] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[76] , wherein one antigen-binding site is formed as a part of the heavy chain / light chain complex constituting the anti-Allergin-1 bispecific antibody;

[79] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[76] , wherein the heavy chain / light chain complex is(a) a combination of a heavy chain having a VH constituting the S1-A-binding site and a light chain having a VL constituting the S1-A-binding site, (b) a combination of a heavy chain having a VH constituting the S1-B-binding site and a light chain having a VL constituting the S1-B-binding site, (c) a combination of a heavy chain having a VH constituting the S2-C-binding site and a light chain having a VL constituting the S2-C-binding site, and (d) a combination of a heavy chain having a VH constituting the S2-D-binding site and a light chain having a VL constituting the S2-D-binding site;

[79] a bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[78] , wherein the other antigen-binding site has the form of an scFv consisting of the VH and VL constituting it;

[80] a bispecific antibody or antibody fragment thereof according to the preceding item

[79] , wherein the scFv is linked to the C-terminus of the heavy chain or light chain;

[81]

[82] The bispecific antibody or antibody fragment thereof according to the preceding item

[79] , wherein the scFv is linked to the C-terminus of the heavy chain;

[83] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[81] , wherein (i) (a) the heavy chain and (b) the light chain are linked to the C-terminus of the scFv, or (ii) the same two heavy chain / light chain complexes are formed from the heavy chain and light chain described in (i) above;

[84] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] ,

[80] , and

[82] , wherein (i) (a) the heavy chain and (b) the light chain are linked to the C-terminus of the scFv, or (ii) the same two heavy chain / light chain complexes are formed from the heavy chain and light chain described in (i) above;

[85] the bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-B binding site, an S2-C binding site, and an S2-D binding site;

[86] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-A binding site, an S2-C binding site, and an S2-D binding site;

[87] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-D binding site;

[88]

[89] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-D binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-C binding site;

[90] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site, and the antigen-binding site in the form of an scFv is an S1-B binding site;

[91]

[92] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site and the antigen-binding site in the form of an scFv is an S2-D binding site;

[93] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site and the antigen-binding site in the form of an scFv is an S2-C binding site;

[94] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site and the antigen-binding site in the form of an scFv is an S2-D binding site;

[95] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site and the antigen-binding site in the form of an scFv is an S1-A binding site;

[96] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site and the antigen-binding site in the form of an scFv is an S1-B binding site;

[97]

[98] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, and the antigen-binding site in the form of an scFv is an S2-D binding site;

[99] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[84] , wherein the antigen-binding site in the heavy chain / light chain complex is an S2-D binding site, and the antigen-binding site in the form of an scFv is an S1-A binding site;

[100]

[101] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[100] , wherein the scFv is linked to the C-terminus of the heavy chain or light chain via (a) a direct amide bond or (b) a peptide linker;

[102] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[79] to

[101] , wherein the N-terminus of VH constituting the scFv is linked to the C-terminus of the heavy chain or light chain via (a) a direct amide bond or (b) a peptide linker;

[103] The bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[79] to

[101] , wherein the N-terminus of the VL constituting the scFv is linked to the C-terminus of the heavy chain or light chain via (a) a direct amide bond or (b) a peptide linker;

[104] The bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[79] to

[102] , wherein the VH and VL constituting the scFv are linked to each other via (a) a direct amide bond or (b) a peptide linker in the order of VH and VL from the N-terminus;

[105] The bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[79] to

[101] and

[103] , wherein the VH and VL constituting the scFv are linked to each other via (a) a direct amide bond or (b) a peptide linker in the order of VL and VH from the N-terminus;

[106]

[107] A bispecific antibody or antibody fragment thereof according to any one of the preceding items

[104] to

[105] , wherein the peptide linker linking the scFv to the C-terminus of the heavy chain or light chain consists of the amino acid sequence of SEQ ID NO: 73;

[108] A bispecific antibody or antibody fragment thereof according to any one of the preceding items

[109] to

[200] , wherein the peptide linker linking the VH and VL constituting the scFv together consists of the amino acid sequence of SEQ ID NO: 74;

[109] A bispecific antibody or antibody fragment thereof having antigen-binding sites that specifically bind to two different epitopes in the extracellular domain of Allergin-1, wherein the antigen-binding sites do not compete with each other for Allergin-1 and are two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site and an S2-D binding site, wherein (I) the S1-A binding site comprises: (i) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and an antigen-binding site consisting of a VL having (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (ii) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28,(b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (iii) an antigen-binding site consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; (iv) any one of antigen-binding sites selected from the group consisting of (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, wherein the S1-B binding site is any one of the antigen-binding sites selected from the group consisting of (v) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; or (vi) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64,(e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, wherein the S2-C binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, wherein the S2-D binding site is a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72; (II) of the two different antigen-binding sites, one is an antigen-binding site in the heavy chain / light chain complex constituting the bispecific antibody, and the other is an antigen-binding site in the form of an scFv; (i) the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-B binding site, an S2-C binding site, and an S2-D binding site; (ii) the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-A binding site, an S2-C binding site, and an S2-D binding site; (iii) the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, and the antigen-binding site in the form of the scFv is any one antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-D binding site; or(iv) the antigen-binding site in the heavy chain / light chain complex is an S2-D binding site, and the antigen-binding site in the form of the scFv is any one antigen-binding site selected from an S1-A binding site, an S1-B binding site, and an S2-C binding site; (III) the N-terminus of either VH or VL constituting the scFv is linked to the C-terminus of the heavy chain or light chain via a peptide linker; (IV) a bispecific antibody or antibody fragment thereof (i) having heavy chains and light chains having the configurations described in the preceding paragraphs (I) to (III), or (ii) consisting of the same two heavy chain / light chain complexes consisting of heavy chains and light chains having the configurations described in the preceding paragraphs (I) to (III);

[109] a bispecific antibody or antibody fragment thereof against Allergin-1, consisting of any two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, (I) An antigen-binding site comprising an S1-A binding site comprising: (i) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (ii) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (iii) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31; (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32; and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55;(e) an antigen-binding site consisting of a VL having a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; and (iv) an antigen-binding site selected from the group consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59 and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60, wherein the S1-B binding site is: (v) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38 and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39; and (d) an antigen-binding site consisting of a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; or (vi) an antigen-binding site consisting of (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, wherein the S2-C binding site is (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45; and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, wherein the S2-D binding site is(a) an antigen-binding site consisting of a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72; (II) one of the two different antigen-binding sites is an antigen-binding site in the heavy chain / light chain complex constituting the bispecific antibody, and the other is an antigen-binding site in the form of an scFv; (i) the antigen-binding site in the heavy chain / light chain complex is an S1-A binding site, and the antigen-binding site in the form of an scFv is any one antigen-binding site selected from an S1-B binding site, an S2-C binding site, and an S2-D binding site; (ii) the antigen-binding site in the heavy chain / light chain complex is an S1-B binding site, and the antigen-binding site in the form of the scFv is any one selected from an S1-A binding site, an S2-C binding site, and an S2-D binding site; (iii) the antigen-binding site in the heavy chain / light chain complex is an S2-C binding site, and the antigen-binding site in the form of the scFv is any one selected from an S1-A binding site, an S1-B binding site, and an S2-D binding site; or (iv) the antigen-binding site in the heavy chain / light chain complex is an S2-D binding site, and the antigen-binding site in the form of the scFv is any one selected from an S1-A binding site, an S1-B binding site, and an S2-C binding site; (III) the N-terminus of either VH or VL constituting the scFv is linked to the C-terminus of the heavy chain or light chain via a peptide linker; (IV) (i) a bispecific antibody or an antibody fragment thereof, which has a heavy chain and a light chain having the configurations described in the preceding paragraphs (I) to (III), or (ii) consists of the same two heavy chain / light chain complexes composed of the heavy chain and the light chain having the configurations described in the preceding paragraphs (I) to (III);

[110] Two different antigen-binding sites are selected from: (i) (a) SEQ ID NOs: 1 and 12, (b) SEQ ID NOs: 2 and 13,(c) an S1-A binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 3 and 14 or 106, (d) SEQ ID NOs: 4 and 15 or 107, (e) SEQ ID NOs: 5 and 16, and (f) SEQ ID NOs: 6 and 17 or 109; (ii) an S1-B binding site composed of a VH and a VL consisting of a pair of amino acid sequences selected from (a) SEQ ID NOs: 7 and 18, or (b) SEQ ID NOs: 8 and 19; (iii) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences selected from SEQ ID NOs: 9 and 20 or 112; and (iv) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences selected from (a) SEQ ID NOs: 10 and 21 or 113, or (b) SEQ ID NOs: 11 and 22;

[111]

[112] A bispecific antibody or antibody fragment thereof according to the preceding item

[108] or

[109] , wherein the two different antigen-binding sites are any two selected from: (i) an S1-A binding site composed of a VH and a VL consisting of any one pair of amino acid sequences selected from (a) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (b) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, (c) SEQ ID NO: 5 and 16, and (d) SEQ ID NO: 6 and SEQ ID NO: 17 or 109, (ii) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112, and (iii) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences of (a) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (b) SEQ ID NO: 11 and 22;

[112] A bispecific antibody or antibody fragment thereof according to the preceding item

[108] or

[109] , wherein the two different antigen-binding sites are any two selected from:

[113] A bispecific antibody or antibody fragment thereof according to the preceding item

[108] or

[109] , wherein the bispecific antibody or antibody fragment thereof is (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 3 and SEQ ID NO: 14 or 106, and (b) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 20 or 112; ...

[114] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the bispecific antibody or antibody fragment thereof is (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 6 and SEQ ID NO: 17 or 109, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 10 and SEQ ID NO: 21 or 113;

[115] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: an S1-A binding site composed of a VH and a VL consisting of any pair of amino acid sequences selected from (a) SEQ ID NO: 3 and SEQ ID NO: 14 or 106, (b) SEQ ID NO: 4 and SEQ ID NO: 15 or 107, and (c) SEQ ID NO: 5 and 16, and (d) SEQ ID NO: 10 and SEQ ID NO: 21 or 113, or (e) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NO: 11 and 22; (a) the S1-A binding site is, (i) when constituting an scFv, the VL constituting the scFv consists of an amino acid sequence selected from SEQ ID NOs: 12 to 17, (ii) when constituting a part of a heavy chain / light chain complex, the VL constituting the light chain consists of an amino acid sequence selected from SEQ ID NOs: 106, 107 and 109, (b) the S1-B binding site is, (i) when constituting an scFv, the VL constituting the scFv consists of the amino acid sequence of SEQ ID NO: 18 or 19, (ii) when constituting a part of a heavy chain / light chain complex, the VL constituting the light chain consists of the amino acid sequence of SEQ ID NO: 110 or 111, (c) the S2-C binding site is, (i) when constituting an scFv, the VL constituting the scFv consists of the amino acid sequence of SEQ ID NO: 20, (ii) when constituting a part of a heavy chain / light chain complex, the VL constituting the light chain consists of the amino acid sequence of SEQ ID NO: 112, (d) the S2-D binding site is: (i) when constituting an scFv, the VL constituting the scFv consists of the amino acid sequence of SEQ ID NO: 21 or 22;(ii) A bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[110] to

[114] , wherein, when the bispecific antibody or antibody fragment thereof constitutes a part of a heavy chain / light chain complex, the VL constituting the light chain consists of the amino acid sequence of SEQ ID NO: 113;

[116] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 3 and 14, and (b) an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 9 and 112;

[117] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences of SEQ ID NOs: 3 and 106, and (b) a bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the bispecific antibody or antibody fragment thereof has an S2-C binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 9 and 20;

[118] a bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 6 and 17, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113;

[119] a bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 6 and 109, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 21;

[120] A bispecific antibody or antibody fragment thereof according to the above item

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 14, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113;

[121] A bispecific antibody or antibody fragment thereof according to the above item

[108] or

[109] , wherein the two different antigen-binding sites are:

[122] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 15, and (b) an S2-D binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 113;

[123] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 3 and 106, and (b) a bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the bispecific antibody or antibody fragment thereof is an S2-D-binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 10 and 21;

[124] a bispecific antibody or antibody fragment thereof according to the preceding paragraph

[108] or

[109] , wherein the two different antigen-binding sites are: (a) an S1-A-binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 4 and 107, and (b) an S2-D-binding site composed of a VH and a VL consisting of a pair of amino acid sequences set forth in SEQ ID NOs: 11 and 22;

[125] a bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[108] to

[124] , wherein the VH and VL constituting the scFv are linked to each other via a peptide linker consisting of the amino acid sequence of SEQ ID NO: 74 in the order of VL and VH from the N-terminus; The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[108] to

[125] , wherein the scFv is linked to the C-terminus of the heavy chain via a peptide linker consisting of the amino acid sequence of SEQ ID NO: 73;

[127] The epitope to which the S1-A binding site specifically binds (S1-A epitope) is composed of at least one or all of phenylalanine at position 70, arginine at position 71, arginine at position 72, lysine at position 73, histidine at position 75, leucine at position 76, histidine at position 96, glutamic acid at position 97, lysine at position 102, lysine at position 104, and tyrosine at position 113 of Allergin-1 having the amino acid sequence represented by SEQ ID NO: 115, and the epitope to which the S2-D binding site specifically binds (S2-D epitope) is composed of at least isoleucine at position 132, leucine at position 133, and tyrosine at position 113 of Allergin-1 having the amino acid sequence represented by SEQ ID NO: 115. a bispecific antibody or an antibody fragment thereof according to any one of the preceding items [1] to

[126] , wherein the bispecific antibody or the antibody fragment thereof is constituted by any one or all of methionine at position 33, valine at position 134, isoleucine at position 135, glutamine at position 136, threonine at position 137, glutamic acid at position 138, histidine at position 146, and proline at position 215, and the epitope to which the S1-B binding site specifically binds is at least an epitope different from the S1-A epitope represented by the amino acids, and the epitope to which the S2-C binding site specifically binds is at least an epitope different from the S2-D epitope represented by the amino acids;

[128] A bispecific antibody or antibody fragment thereof against Allergin-1, comprising two different antigen-binding sites selected from an S1-A binding site, an S1-B binding site, an S2-C binding site, and an S2-D binding site, wherein: (a) the epitope to which the S1-A binding site specifically binds (S1-A epitope) is composed of at least one or all of phenylalanine at position 70, arginine at position 71, arginine at position 72, lysine at position 73, histidine at position 75, leucine at position 76, histidine at position 96, glutamic acid at position 97, lysine at position 102, lysine at position 104, and tyrosine at position 113 of Allergin-1 comprising the amino acid sequence set forth in SEQ ID NO: 115;(b) the epitope to which the S2-D-binding site specifically binds (S2-D epitope) is composed of at least one or all of isoleucine at position 132, methionine at position 133, valine at position 134, isoleucine at position 135, glutamine at position 136, threonine at position 137, glutamic acid at position 138, histidine at position 146, and proline at position 215 of Allergin-1; (c) the epitope to which the S1-B-binding site specifically binds is at least an epitope different from the S1-A epitope represented by the aforementioned amino acids; and (d) the epitope to which the S2-C-binding site specifically binds is at least an epitope different from the S2-D epitope represented by the aforementioned amino acids;

[129]

[130] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[77] to

[128] , wherein the heavy chain and the light chain are both derived from an IgG antibody;

[131] The bispecific antibody or antibody fragment thereof according to the preceding item

[129] , wherein the heavy chain has CH1, CH2, and CH3 of an IgG antibody, and the light chain has CL of the same antibody;

[132] The IgG antibody is an IgG １ Antibodies or IgG ４

[132] The bispecific antibody or antibody fragment thereof according to the above

[129] or

[130] , which is an IgG antibody;

[133] The bispecific antibody or antibody fragment thereof according to the above

[129] or

[130] , which is an IgG antibody; １ The bispecific antibody or antibody fragment thereof according to the above item

[129] or

[130] , which is an antibody;

[133] the IgG １

[134] The bispecific antibody or antibody fragment thereof according to the above item

[132] , wherein the leucine at position 235 in the two heavy chain constant regions of the antibody is each substituted with glycine and / or the glycine at position 236 is each substituted with arginine, according to the EU numbering system;

[135] The IgG １

[135] The bispecific antibody or antibody fragment thereof according to the above

[132] or

[133] , wherein the lysine at position 447 according to the EU numbering system in each of the two heavy chain constant regions of the antibody is missing;

[136] The IgG antibody or antibody fragment thereof according to the above

[136] or

[137] , wherein the IgG antibody is an IgG ４

[136] The bispecific antibody or antibody fragment thereof according to the above item

[129] or

[130] , which is an antibody;

[137] The IgG４

[137] A bispecific antibody or antibody fragment thereof according to the preceding paragraph

[77] to

[132] , wherein the heavy chain constant region consists of the amino acid sequence of SEQ ID NO: 23 or 99;

[138] A bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[77] to

[132] and

[137] , wherein the light chain constant region consists of the amino acid sequence of SEQ ID NO: 24;

[139] A bispecific antibody or antibody fragment thereof according to any one of the preceding paragraphs

[77] to

[132] , wherein the heavy chain constant region and the light chain constant region consist of the amino acid sequence of SEQ ID NO: 23 or 99 and SEQ ID NO: 24, respectively;

[140]

[141] The bispecific antibody or antibody fragment thereof according to any one of the preceding items

[137] to

[139] , wherein the heavy chain constant region consists of the amino acid sequence of SEQ ID NO: 23 or 99, in which arginine at position 97 in the amino acid sequence of SEQ ID NO: 23 or 99 is substituted with lysine;

[142] (i) (a) SEQ ID NO: 75 and 76, (b) SEQ ID NO: 77 and 78, (c) SEQ ID NO: 79 and 78, (d) SEQ ID NO: 80 and 78, (e) SEQ ID NO: 81 and 82, (f) SEQ ID NO: 83 and 82, (g) SEQ ID NO: 84 and 85, (h) SEQ ID NO: 86 and 87, (i) SEQ ID NO: 88 and 89, (j) SEQ ID NO: 90 and 91, (k) SEQ ID NO: 90 and 92, (l) SEQ ID NO: 93 and 94, (m) SEQ ID NO: 93 and 95, (n) SEQ ID NO: 96 and 97, (o) SEQ ID NO: 98 and 78, (p) SEQ ID NOs: 100 and 78, (q) SEQ ID NOs: 101 and 78, (r) SEQ ID NOs: 102 and 78, and (s) SEQ ID NOs: 103 and 85, or (ii) a bispecific antibody against Allergin-1, comprising the same two heavy chain / light chain complexes composed of the heavy chain and light chain described in the preceding paragraph (i);

[142] (i) (a) SEQ ID NOs: 77 and 78, (b) SEQ ID NOs: 79 and 78, (c) SEQ ID NOs: 83 and 82, (d) SEQ ID NOs: 84 and 85, (e) SEQ ID NOs: 98 and 78, (f) SEQ ID NOs: 100 and 78,(g) a bispecific antibody against Allergin-1, having heavy and light chains consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 101 and 78, (h) SEQ ID NOs: 102 and 78, and (i) SEQ ID NOs: 103 and 85, or (ii) consisting of the same two heavy chain / light chain complexes consisting of the heavy and light chains according to the preceding paragraph (i);

[143] a bispecific antibody against Allergin-1 according to the preceding paragraph

[141] or

[142] , in which arginine at position 214 according to the EU numbering system in the constant region of the heavy chain has been substituted with lysine, and / or alanine at position 299 in SEQ ID NOs: 75, 81, 83, 86, 88, 93, and 96 and at position 301 in SEQ ID NOs: 80 and 90 (corresponding to position 297 according to the EU numbering system in the constant region of the heavy chain) has been substituted with asparagine;

[144]

[145] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[143] , wherein cytokine production during administration or within 24 hours after administration is sufficiently reduced;

[145] The bispecific antibody or antibody fragment thereof according to the preceding item

[144] , wherein the cytokine comprises at least one or more cytokines selected from IL-2, IL-4, IL-6, IL-10, IFN-γ, and TNF-α;

[146] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[145] , wherein the bispecific antibody or antibody fragment thereof inhibits binding of a TLR9 ligand (preferably, ODN M362, ODN 1668, ODN 1826, etc.) to Allergin-1;

[147] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[146] , wherein Allergin-1 is human Allergin-1;

[148]

[149] The bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[148] , which is a monoclonal antibody;

[150] An isolated antibody,[1-1] A bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[149] ; [1-1] A pharmaceutical composition comprising, as an active ingredient, the anti-Allergin-1 bispecific antibody or antibody fragment thereof according to any one of the preceding items [1] to

[150] ; [1-2] The pharmaceutical composition according to the preceding item [1-1], further comprising at least one pharmaceutically acceptable carrier; [2-1] An agent for suppressing the progression of symptoms, suppressing recurrence and / or treating cancer, comprising, as an active ingredient, an anti-Allergin-1 bispecific antibody or antibody fragment thereof selected from any one of the preceding items [1] to

[150] ; [2-2] The agent according to the preceding item [2-1], wherein the cancer is a solid cancer or a blood cancer; [2-3] The cancer is a solid cancer, and the solid cancer is selected from the group consisting of malignant melanoma (e.g., malignant melanoma in the skin, oral mucosal epithelium, or orbit, etc.), non-small cell lung cancer (e.g., squamous non-small cell lung cancer and non-squamous non-small cell lung cancer), small cell lung cancer, head and neck cancer (e.g., oral cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, salivary gland cancer, and tongue cancer), renal cell carcinoma (e.g., clear cell renal cell carcinoma), breast cancer, ovarian cancer (e.g., serous ovarian cancer and ovarian clear cell adenocarcinoma), uterine cancer (e.g., cervical cancer and endometrial cancer), anal cancer (e.g., anal canal cancer), colorectal cancer (e.g., MSI-H and / or dMMR-positive colorectal cancer), rectal cancer, colon cancer, hepatocellular carcinoma, esophageal cancer, gastric cancer, esophagogastric junction cancer, pancreatic cancer ( the agent according to the preceding item [2-2], which is for example one or more cancers selected from pancreatic ductal carcinoma, insulinoma, intraductal papillary mucinous tumor), urothelial cancer (e.g., bladder cancer, upper urinary tract cancer, ureteral cancer, renal pelvic cancer, and urethral cancer), prostate cancer, fallopian tube cancer, primary peritoneal cancer, malignant pleural mesothelioma, gallbladder cancer, bile duct cancer, biliary tract cancer, skin cancer (e.g., uveal melanoma and Merkel cell carcinoma), testicular cancer (germ cell tumor), vaginal cancer, vulvar cancer, penile cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, spinal tumor, neuroblastoma, medulloblastoma, ocular retinoblastoma, neuroendocrine tumor, brain tumor (e.g., glioma (e.g., glioblastoma and gliosarcoma) and meningioma) and squamous cell carcinoma; [2-4] The agent according to the preceding item [2-2], wherein the cancer is a solid cancer, and the solid cancer is bone and soft tissue sarcoma (e.g., Ewing's sarcoma, childhood rhabdomyosarcoma, uterine leiomyosarcoma, chondrosarcoma, pulmonary sarcoma, osteosarcoma, and congenital fibrosarcoma) or Kaposi's sarcoma; [2-5] The agent according to the preceding item [2-2], wherein the cancer is a blood cancer, and the blood cancer is multiple myeloma,Malignant lymphomas (e.g., non-Hodgkin's lymphomas (e.g., B-cell non-Hodgkin's lymphomas (e.g., precursor B-cell lymphoblastic lymphoma, precursor B-cell acute lymphoblastic leukemia, chronic B-lymphocytic leukemia, B-cell precursor leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, nodal marginal zone B-cell lymphoma, extranodal marginal zone B-cell lymphoma (MALT lymphoma), primary splenic marginal zone B-cell lymphoma, hairy cell leukemia, hairy cell leukemia variant, follicular lymphoma, pediatric follicular lymphoma, diffuse large B-cell lymphoma, diffuse large B-cell lymphoma, Cell lymphoma, unspecified type, splenic diffuse red pulp small B-cell lymphoma, primary mediastinal large B-cell lymphoma, primary effusion lymphoma, Burkitt lymphoma, mantle cell lymphoma, monoclonal B-cell lymphocytosis, splenic B-cell lymphoma / leukemia, unclassifiable type, monoclonal gammopathy of undetermined significance, IgM type, μ heavy chain disease, λ heavy chain disease, α heavy chain disease, plasma cell myeloma, solitary plasmacytoma of bone, extraskeletal plasmacytoma, monoclonal immunoglobulin deposition disease, large B-cell lymphoma with IRF4 rearrangement, primary cutaneous follicle center lymphoma, T-cell / histiocytic abundant B-cell-rich large B-cell lymphoma, primary central nervous system diffuse large B-cell lymphoma, primary cutaneous diffuse large B-cell lymphoma - leg type, EBV-positive diffuse large B-cell lymphoma - not otherwise specified, EBV-positive mucocutaneous ulcer, chronic inflammation-associated diffuse large B-cell lymphoma, lymphomatoid granulomatosis, intravascular large B-cell lymphoma, ALK-positive large B-cell lymphoma, plasmablastic lymphoma, HHV8-positive diffuse large B-cell lymphoma - not otherwise specified, Burkitt-like lymphoma with 11q aberration, MYC and BCL2 and / or BCL6 rearrangements high-grade B-cell lymphoma with features intermediate between diffuse large B-cell lymphoma and classical Hodgkin lymphoma, high-grade B-cell lymphoma, unspecified type and diffuse large B-cell lymphoma, unspecified type), T / NK cell non-Hodgkin lymphoma (e.g., precursor T-cell lymphoblastic lymphoma, chronic T-lymphocytic leukemia, T-cell acute lymphoblastic leukemia, T-cell large granular lymphocytic leukemia, large granular NK cell leukemia, aggressive NK cell leukemia, peripheral T-cell lymphoma, peripheral T-cell lymphoma, unspecified type, peripheral T-cell lymphoma, unspecified type, angioimmunoblastic T-cell lymphoma,Anaplastic large cell (CD30 positive) lymphoma, angiocentric lymphoma, intestinal T-cell lymphoma, enteropathic T-cell lymphoma, hepatosplenic gamma-delta T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, mycosis fungoides, Sézary syndrome, Hodgkin-like / Hodgkin-related anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma, adult T-cell lymphoma, T-cell prolymphocytic leukemia, chronic NK-cell lymphoproliferative disorder, pediatric systemic EBV-positive T-cell lymphoma, smallpox vaccination bullosa-like lymphoproliferative disorder, extranodal NK / T-cell lymphoma / nasal type, enteropathy-associated T-cell lymphoma, monomorphic epitheliotropic intestinal T-cell lymphoma, gastrointestinal slowly progressive T-cell lymphoproliferative disorder, hepatosplenic T-cell lymphoma, primary skin CD30-positive T-cell lymphoproliferative disorder, lymphomatoid papulopathy, primary cutaneous anaplastic large-cell lymphoma, cutaneous T-cell lymphoma, primary cutaneous γδ T-cell lymphoma, primary cutaneous CD8-positive rapidly progressive epidermotropic cytotoxic T cells cutaneous acral CD8-positive T-cell lymphoma, primary cutaneous CD4-positive small / medium T-cell lymphoproliferative disorder, follicular T-cell lymphoma, nodal peripheral T-cell lymphoma with follicular helper T-cell phenotype, anaplastic large cell lymphoma ALK-positive, anaplastic large cell lymphoma ALK-negative, and breast implant-associated anaplastic large cell lymphoma) and Hodgkin lymphoma (e.g., classical Hodgkin lymphoma (e.g., nodular sclerosing, mixed cytology) the agent according to the preceding item [2-2], wherein the cancer is one or more selected from the group consisting of leukemia (e.g., acute myeloid leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia (lymphoblastic lymphoma), chronic lymphocytic leukemia (small lymphocytic lymphoma), myelodysplastic syndrome, and chronic myeloid leukemia), primary central nervous system malignant lymphoma, and myeloproliferative syndrome; [2-6] The agent according to any one of the preceding items [2-1] to [2-5], wherein the cancer is childhood cancer or cancer of unknown primary; [2-7] The agent according to any one of the preceding items [2-1] to [2-6], wherein the cancer is a cancer for which the therapeutic effect of other anti-cancer agents is insufficient or insufficient; [2-8] The agent according to any one of the preceding items [2-1] to [2-7], wherein the cancer has progressed after treatment with other anti-cancer agents; [2-9] The cancer patient is a patient with no history of treatment with other anti-cancer agents,[2-10] The agent according to any one of the preceding items [2-1] to [2-9], which is prescribed for postoperative adjuvant therapy or neoadjuvant therapy; [2-11] The agent according to any one of the preceding items [2-1] to [2-10], wherein the cancer is curable or inresectable, metastatic, recurrent, refractory, and / or distant metastatic; [2-12] The agent according to any one of the preceding items [2-1] to [2-11], wherein the TPS or CPS is 50% or more, 25% or more, 10% or more, 5% or more, or 1% or more; [2-13] The agent according to any one of the preceding items [2-1] to [2-11], wherein the TPS or CPS is less than 50%, less than 25%, less than 10%, less than 5%, or less than 1%; [2-14] [2-15] The agent according to any one of the preceding items [2-1] to [2-13], wherein the incidence of TMB is high; [2-16] The agent according to any one of the preceding items [2-1] to [2-15], wherein the incidence of TMB is low; [2-17] The agent according to any one of the preceding items [2-1] to [2-15], further characterized by being used in combination with another anti-cancer agent; [2-18] The agent according to any one of the preceding items [2-7] to [2-9] and [2-16], wherein the other antineoplastic agent is one or more drugs selected from alkylating agents, platinum preparations, antimetabolites (e.g., antifolates, pyridine metabolism inhibitors, and purine metabolism inhibitors), ribonucleotide reductase inhibitors, nucleotide analogs, topoisomerase inhibitors, microtubule polymerization inhibitors, microtubule depolymerization inhibitors, antitumor antibiotics, cytokine preparations, antihormones, molecularly targeted drugs, and tumor immunotherapeutic drugs (preferably drugs that are not likely to cause cytokine release syndrome); the other anti-cancer agent is a molecular targeted drug, and the molecular targeted drug is an ALK inhibitor, a BCR-ABL inhibitor, an EGFR inhibitor, a B-RAF inhibitor, a VEGFR inhibitor, an FGFR inhibitor, a c-MET inhibitor, an AXL inhibitor, a MEK inhibitor, a CDK inhibitor, a BTK inhibitor, a BCL-2 inhibitor, a PI3K-δ / γ inhibitor, a JAK-1 / 2 inhibitor, an ERK inhibitor, a TGFbR1 inhibitor, a cancer cell stemness kinase inhibitor, a FAK inhibitor, a SYK / FLT3 dual inhibitor, an ATR inhibitor, a WEE1 kinase inhibitor, a multi-tyrosine kinase inhibitor, an mTOR inhibitor, an HDAC inhibitor, a PARP inhibitor,アロマターゼ inhibitor, EZH2 inhibitor, ガレクチン-3 inhibitor, STAT3 inhibitor, DNMT inhibitor, SMO inhibitor, HSP90 inhibitor, γ-チューブリンSpecific inhibitors, HIF2α inhibitors, グルタミナーゼ inhibitors, E3 リガーゼ inhibitors, NRF2 activating agents, アルギナーゼ inhibitors, cell cycle inhibitors, Ephrin B4 inhibitors, IAP antagonists, anti-HER2 antibodies, anti-HER1 antibodies, anti-HER3 antibodies, anti-CD40 antibodies, anti-CD70 antibodies, anti-VEGF antibodies, anti-VEGFR1 antibodies, anti-VEGFR2 antibodies, anti-CD20 antibodies, anti-CD30 antibodies, anti-CD38 antibodies, anti-TNFRSF10B antibodies, anti-TNFRSF10A antibodies, anti-MUC1 antibodies, anti-MUC5AC antibodies, anti-MUC16 antibodies, anti-DLL4 antibodies, anti-FukosylGM1 antibodies, anti-gpNMB antibodies, anti-Mesothelin antibodies Antibodies, anti-MMP9 antibodies, anti-GD2 antibodies, anti-MET antibodies, anti-FOLR1 antibodies, anti-CD79b antibodies, anti-DLL3 antibodies, anti-CD51 antibodies, anti-EPCAM antibodies, anti-CEACAM5 antibodies, anti-CEACAM6 antibodies, anti-FGFR2 antibodies, anti-CD44 antibodies, anti-PSMA antibodies, anti-Endoglin antibodies, anti-IGF1R antibodies, anti-TNFSF11 antibodies, anti-GUCY2C antibodies, anti-SLC39A6 antibodies, anti-SLC34A2 antibodies, anti-NCAM1 antibodies, anti-ganglioside GD3 antibody, anti-AMHR2 antibody, anti-CD37 antibody, anti-IL1RAP antibody, anti-PDGFR2 antibody, anti-CD200 antibody, anti-TAG-72 antibody, anti-SLITRK6 antibody, anti-DPEP3 antibody, anti-CD19 antibody, anti-NOTCH2 / 3 antibody, anti-tenascin C antibody, anti-AXL antibody, anti-STEAP1 antibody, anti-CTAA16 antibody, anti-CLDN18 antibody, anti-GM3 antibody, anti-PSCA antibody, anti-FN extra domain antibody B antibody, anti-HAVCR1 antibody, anti-TNFRSF4 antibody, anti-HER1-MET dual-specificity antibody, anti-EPCAM-CD3 dual-specificity antibody, anti-Ang2-VEGF dual-specificity antibody, anti-HER2-CD3 dual-specificity antibody, anti-HER3-IGF1R dual-specificity antibody, anti-PMSA-CD3 dual-specificity antibody, anti-HER1-LGR5 dual-specificity antibody, anti-SSTR2-CD3 dual-specificity antibody, anti-CD30-CD16A dual-specificity antibody, anti-CEA-CD3 dual-specificity antibody, anti-CD3-CD19 dual-specificity antibody, anti-IL3RA-CD3 dual-specificity antibody, anti-GPRC5D-CD3 dual-specificity antibody, anti-CD20-CD3 dual-specificity antibody,The agent according to the above item [2-17], which is any one drug selected from an anti-TNFRSF17-CD3 bispecific antibody, an anti-CLEC12A-CD3 bispecific antibody, an anti-HER2-HER3 bispecific antibody, an anti-FAP antibody / IL-2 fusion protein, and an anti-CEA antibody / IL-2 fusion protein; [2-19] The other anti-cancer agent is a tumor immunotherapy agent, and the tumor immunotherapy agent is an anti-PD-1 antibody, anti-PD-L1 antibody, PD-1 antagonist, PD-L1 / VISTA antagonist, PD-L1 / TIM3 antagonist, anti-PD-L2 antibody, PD-L1 fusion protein, PD-L2 fusion protein, anti-CTLA-4 antibody, anti-LAG-3 antibody, anti-TIM3 antibody, anti-KIR antibody, anti-BTLA antibody, anti-TIGIT antibody, anti-VISTA antibody, anti-CD137 antibody, anti-CSF-1R antibody / CSF-1R inhibitor, anti-OX40 antibody, anti-HVEM antibody, anti-CD27 antibody, anti-GITR antibody / the agent according to the preceding item [2-17], which is any one drug selected from a GITR fusion protein, an anti-CD28 antibody, an anti-CCR4 antibody, an anti-B7-H3 antibody, an anti-ICOS agonist antibody, an anti-CD4 antibody, an anti-DEC-205 antibody / NY-ESO-1 fusion protein, an anti-SLAMF7 antibody, an anti-CD73 antibody, PEGylated IL-2, an IDO inhibitor, a TLR agonist, an adenosine A2A receptor antagonist, an anti-NKG2A antibody, an anti-CSF-1 antibody, an immunopotentiator, an IL-15 superagonist, soluble LAG3, an anti-CD47 antibody / CD47 antagonist, and an IL-12 antagonist; [2-20] The agent according to the preceding item [2-17], wherein the other anti-malignant tumor agent is a tumor immunotherapeutic agent, and the tumor immunotherapeutic agent is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody; [2-21] The tumor immunotherapeutic agent is an anti-PD-1 antibody, and the anti-PD-1 antibody is selected from the group consisting of Nivolumab, Cemiplimab-rwlc, Pembrolizumab, Spartalizumab, Tislelizumab, Dostarlimab, Toripalimab, Camrelizumab, Genolimzumab, Sintilimab, Lodapolimab, Retifanlimab, Balstilimab, Serplulimab, Budigalimab, Prolgolimab, Sasanlimab, Cetrelimab, Zimberelimab, Geptanolimab, AMP-514,The agent according to the above item [2-20], which is an antibody selected from STI-A1110, ENUM 388D4, ENUM 244C8, Zimberelimab, CS1003, BAT-1306, Penpulimab, AK103, Ezabenlimab, LZM009, CMAB819, Sym021, SSI-361, JY034, Pucotenlimab, ISU106, and CX-188; [2-22] [2-23] The agent according to the preceding item [2-20], wherein the tumor immunotherapeutic agent is an anti-PD-L1 antibody selected from Atezolizumab, Avelumab, Durvalumab, Manelimab, Pacmilimab, Envafolimab, Cosibelimab, Sugemalimab, BMS-936559, STI-1014, HLX20, Adebrelimab, MSB2311, BGB-A333, KL-A167, AK106, Socazolimab, FAZ053, CBT-502, and JS003; [2-24] The agent according to the preceding item [2-20], wherein the tumor immunotherapeutic agent is an anti-CTLA-4 antibody selected from Ipilimumab, Zalifrelimab, Nurulimab, and Tremelimumab; [2-25] The agent described in the preceding item [2-21], when the anti-PD-1 antibody is nivolumab, to be administered to adults by intravenous infusion at the following doses: (a) 1 mg / kg (body weight) of nivolumab every 3 weeks, (b) 3 mg / kg (body weight) of nivolumab every 2 weeks, (c) 2 mg / kg (body weight) of nivolumab every 3 weeks, (d) 80 mg of nivolumab every 3 weeks, (e) 240 mg of nivolumab every 2 weeks, (f) 360 mg of nivolumab every 3 weeks, or (g) 480 mg of nivolumab every 4 weeks; [2-26] The agent described in the preceding item [2-26], when the anti-PD-1 antibody is pembrolizumab, to be administered to adults by intravenous infusion at the following doses: (a) 200 mg of nivolumab every 3 weeks, (b) 400 mg of nivolumab every 6 weeks, or (c) 200 mg of nivolumab every 3 weeks, (b) 400 mg of nivolumab every 6 weeks, or (d) 200 mg of nivolumab every 3 weeks, (c) 200 mg of nivolumab every 2 weeks, (d) 200 mg of nivolumab every 2 weeks, (e) 240 mg of nivolumab every 2 weeks, (f) 360 mg of nivolumab every 3 weeks, or (g) 480 mg of nivolumab every 4 weeks; the agent described in the preceding item [2-21], which is administered by intravenous drip infusion at a dose of 200 mg / kg (body weight) (up to a maximum of 200 mg per administration) every 3 weeks; [2-26] when the anti-PD-1 antibody is cemiplimab-rwlc,[2-27] The agent described in the preceding paragraph [2-21], wherein to adults, 350 mg of Cemiplimab-rwlc is administered by intravenous infusion every 3 weeks; [2-28] The agent described in the preceding paragraph [2-22], wherein to adults, 10 mg / kg (body weight) of Avelumab is administered by intravenous infusion every 2 weeks; [2-29] The agent described in the preceding paragraph [2-29], wherein to adults, 10 mg of Atezolizumab is administered by intravenous infusion every 2 weeks; [2-29] The agent described in the preceding paragraph [2-29], wherein to adults, 10 mg of Atezolizumab is administered by intravenous infusion every 2 weeks; [2-30] The agent described in the preceding paragraph [2-31], wherein to adults, 500 mg of Dostarlimab is administered by intravenous infusion every 3 weeks, followed by 1 to 4 doses of Dostarlimab every 3 weeks, followed by 1,000 mg of Dostarlimab every 6 weeks; [2-31] (b) the agent described in the preceding paragraph [2-22], administered by intravenous infusion at a dose of 1200 mg every three weeks, or (c) 1680 mg every four weeks; [2-30] the agent described in the preceding paragraph [2-22], wherein the anti-PD-L1 antibody is durvalumab, and the agent is administered to adults by intravenous infusion at a dose of 10 mg / kg (body weight) of durvalumab every two weeks; [2-31] the agent described in the preceding paragraph [2-23], wherein the anti-CTLA-4 antibody is ipilimumab, and the agent is administered to adults by intravenous infusion at a dose of (a) 3 mg / kg (body weight) of ipilimumab once daily, or (b) 1 mg / kg (body weight) of ipilimumab once daily, every three weeks for four doses; [2-32] The agent according to any one of the preceding items [2-20] to [2-31], wherein the anti-PD-1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody is administered by intravenous infusion over 30 minutes, 60 minutes, 30 to 60 minutes, or more than 60 minutes; [3-1] An intravenous preparation comprising an anti-Allergin-1 bispecific antibody or antibody fragment thereof selected from any one of the preceding items [1] to

[150] and at least one pharmaceutically acceptable carrier; [3-2] An intravenous preparation according to the preceding item [3-1] for inhibiting the progression of symptoms, inhibiting recurrence, and / or treating cancer; [3-3] An intravenous preparation for infusion,[4-1] A method for suppressing the progression of symptoms, suppressing recurrence and / or treating cancer, comprising administering to a patient an effective amount of an anti-Allergin-1 bispecific antibody or antibody fragment thereof selected from any one of the preceding items [1] to

[150] ; [5-1] An anti-Allergin-1 bispecific antibody or antibody fragment thereof selected from any one of the preceding items [1] to

[150] , for use in suppressing the progression of symptoms, suppressing recurrence and / or treating cancer; [6-1] Use of an anti-Allergin-1 bispecific antibody or antibody fragment thereof selected from any one of the preceding items [1] to

[150] in the manufacture of a medicament for suppressing the progression of symptoms, suppressing recurrence and / or treating cancer; [7-1] (a) SEQ ID NOs: 1 and 104, (b) SEQ ID NOs: 2 and 105, (c) SEQ ID NOs: 3 and 106, (d) SEQ ID NOs: 4 and 107, (e) SEQ ID NOs: 5 and 108, (f) SEQ ID NOs: 6 and 109, (g) SEQ ID NOs: 7 and 110, and (h) SEQ ID NOs: 8 and 111, and specifically binds to the S1 domain of Allergin-1 (anti-S1 domain antibody) or an antibody fragment thereof, which cross-competes with the binding to the S1 domain of Allergin-1 of an antibody or antigen-binding site thereof having VH and VL consisting of any one pair of amino acid sequences selected from: (a) SEQ ID NOs: 1 and 104, (b) SEQ ID NOs: 2 and 105, (c) SEQ ID NOs: 3 and 106, (d) SEQ ID NOs: 4 and 107, (e) SEQ ID NOs: 5 and 108, (f) SEQ ID NOs: 6 and 109, (g) SEQ ID NOs: 7 and 110, and (h) an antibody or antibody fragment thereof that cross-competes with an antibody or antigen-binding site thereof having a VH and VL consisting of any one pair of amino acid sequences selected from SEQ ID NOs: 8 and 111; [7-3] an antibody or antibody fragment thereof that cross-competes with an antibody or antigen-binding site thereof having a VH and VL consisting of any one pair of amino acid sequences selected from (a) SEQ ID NOs: 9 and 112, (b) SEQ ID NOs: 10 and 113, and (c) SEQ ID NOs: 11 and 114 for binding to the S2 domain of Allergin-1;An antibody (anti-S2 domain antibody) or an antibody fragment thereof that specifically binds to the S2 domain of Allergin-1; [7-4] An anti-S2 domain antibody or an antibody fragment thereof, wherein the binding of the antibody or antibody fragment thereof to the S2 domain of Allergin-1 is cross-competed with an antibody or an antigen-binding site thereof having a VH and a VL consisting of any one pair of amino acid sequences selected from (a) SEQ ID NOs: 9 and 112, (b) SEQ ID NOs: 10 and 113, and (c) SEQ ID NOs: 11 and 114; [7-5] (A) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 26, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51; (B) (a) a VH having a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 29, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54; (C) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 32, and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56, and (f) a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57; (D) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34; (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35; and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36;and (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 60; (E) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 38, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63; and (F) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 41, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42; (d) a VL having a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66; [7-6] (A) a VH having (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44, and (c) a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) a VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68, and (f) a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, or (B) (a) a VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) a VH having a VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47; and (c) a VH having a VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48; and (d) a VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70.(e) the antibody or antibody fragment thereof according to the preceding paragraph [7-3] or [7-4], which has a pair of VH and VL, wherein VL has VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71 and (f) VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72; [7-7] the antibody or antibody fragment thereof according to the preceding paragraph [7-5] or [7-6], in which any 1 to 5 amino acid residues in any one or more CDRs selected from VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2 and VL-CDR3 may be substituted with other amino acids; [7-8] the antibody or antibody fragment thereof according to any one of the preceding paragraphs [7-1] to [7-7], in which FW of VH and VL consists of amino acid sequences encoded by germline VH gene IGHV3-66 and VL gene IGKV1-39, respectively, or the genes thereof that have undergone one or more somatic mutations and / or back mutations; [7-9] The antibody or antibody fragment thereof according to any one of the preceding items [7-1], [7-2], [7-5], [7-7] and [7-8], having a VH and a VL consisting of any one set of amino acid sequences selected from (a) SEQ ID NOs: 1 and 104, (b) SEQ ID NOs: 2 and 105, (c) SEQ ID NOs: 3 and 106, (d) SEQ ID NOs: 4 and 107, (e) SEQ ID NOs: 5 and 108, (f) SEQ ID NOs: 6 and 109, (g) SEQ ID NOs: 7 and 110, and (h) SEQ ID NOs: 8 and 111, or a set of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of the amino acid sequences (excluding the three CDRs of each); [7-10] (a) SEQ ID NOs: 9 and 112, (b) SEQ ID NOs: 10 and 113, and (c) the antibody or antibody fragment thereof according to any one of the preceding items [7-3], [7-4], and [7-6] to [7-8], having VH and VL consisting of any pair of amino acid sequences selected from SEQ ID NOs: 11 and 114, or a pair of amino acid sequences that are independently at least 80%, 90%, 95%, 98%, or 99% identical to each of said amino acid sequences (excluding three CDRs for each); [7-11] an S1-A binding site, an S1-B binding site,An antibody or antibody fragment thereof against Allergin-1, having an antigen-binding site selected from an S2-C binding site and an S2-D binding site, wherein: (a) the epitope to which the S1-A binding site specifically binds (S1-A epitope) is composed of at least one or all of phenylalanine at position 70, arginine at position 71, arginine at position 72, lysine at position 73, histidine at position 75, leucine at position 76, histidine at position 96, glutamic acid at position 97, lysine at position 102, lysine at position 104, and tyrosine at position 113 of Allergin-1 consisting of the amino acid sequence represented by SEQ ID NO: 115; (b) the epitope to which the S2-D binding site specifically binds (S2-D epitope) is composed of at least one or all of isoleucine at position 132, methionine at position 133, valine at position 134, isoleucine at position 135, glutamine at position 136, threonine at position 137, glutamic acid at position 138, histidine at position 146, and proline at position 215 of the Allergin-1; (c) the epitope to which the S1-B binding site specifically binds is at least an epitope different from the S1-A epitope represented by the amino acids; and (d) the epitope to which the S2-C binding site specifically binds is at least an epitope different from the S2-D epitope represented by the amino acids; [7-12] the antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-11], which is an IgG antibody; [7-13] The IgG antibody is IgG, １ Antibodies or IgG ４[7-14] The antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-13], wherein the Allergin-1 is human Allergin-1; [7-15] The antibody or antibody fragment thereof according to the preceding item [7-14], wherein the human Allergin-1 is any one or more of human Allergin-1 selected from human Allergin-1L, human Allergin-1S1, and human Allergin-1S2; [7-16] The antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-7] and [7-11] to [7-15], wherein the antibody or antibody fragment thereof is a rabbit antibody; [7-17] The antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-7] and [7-11] to [7-16], wherein the antibody or antibody fragment thereof is a chimeric antibody; [7-18] [7-19] The antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-18], which is a monoclonal antibody; [7-20] The antibody or antibody fragment thereof according to any one of the preceding items [7-1] to [7-19], which is an isolated antibody; [8-1] A polynucleotide encoding any of the amino acid sequences selected from SEQ ID NOs: 1 to 24 and SEQ ID NOs: 75 to 114; [8-2] An expression vector comprising any of the polynucleotides according to the preceding item [8-1]; and [8-3] An animal cell into which the expression vector according to the preceding item [8-2] has been introduced or which has been transformed by introducing said vector.

[0122] The contents of all patent and non-patent literature or references explicitly cited in this specification are hereby incorporated by reference in their entirety.

[0123] The present invention will be described in more detail by the following examples, but the scope of the present invention is not limited thereto. Various changes and modifications can be made by those skilled in the art based on the description of the present invention, and these changes and modifications are also included in the present invention.

[0124] Example 1: Immunization of rabbits with recombinant human Allergin-1L-His fusion protein and human Allergin-1L expression plasmid vector To obtain antibodies (including anti-S1 domain antibodies and anti-S2 domain antibodies) that specifically bind to human Allergin-1 of the present invention, rabbits were immunized with recombinant human Allergin-1L protein. Similarly, rabbits were immunized with expression vectors containing cDNAs encoding human Allergin-1L, human Allergin-1S1, and human Allergin-1S2, respectively.

[0125] Nine rabbits were immunized with recombinant human Allergin-1L protein by subcutaneous injection four times over 56 days or five times over 104 days with recombinant human Allergin-1L-His fusion protein (Sino Biological, product number 16083-H08H) emulsified with incomplete Freund's adjuvant or complete Freund's adjuvant. Blood samples were collected from the immunized rabbits, and serum antibody titers were assessed by ELISA using the recombinant human Allergin-1L-His fusion protein or by Cell ELISA (CELISA) using a CHO cell line stably expressing human Allergin-1L.

[0126] Eleven rabbits were immunized with expression vectors containing cDNA encoding human Allergin-1 (Human Allergin-1L, Human Allergin-1S1, and Human Allergin-1S2) subcutaneously eight times over 114 days. Blood samples were collected from the immunized rabbits, and serum antibody titers were assessed by CELISA using CHO cell lines stably expressing human Allergin-1L, human Allergin-1S1, and human Allergin-1S2.

[0127] Spleens were removed from individuals with confirmed elevated serum antibody titers, and lymphocytes were prepared from the spleens. The percentage of cell populations that bound fluorescently labeled recombinant human Allergin-1L-His fusion protein was analyzed by flow cytometry, and the percentage of antigen-specific antibody-producing B cells was calculated. Splenic lymphocytes from individuals with high serum antibody titers and high percentages of antigen-specific antibody-producing B cells among splenic lymphocytes were used to select anti-Allergin-1 antibody-producing B cells.

[0128] Example 2: Selection of B cells producing anti-Allergin-1 antibodies. Cells that bound fluorescently labeled recombinant human Allergin-1L-His fusion protein, i.e., B cells producing antigen-specific antibodies, were isolated by flow cytometry from splenic lymphocytes collected from each individual rabbit selected in Example 1. The isolated B cells were cultured, and the binding of antibodies in the culture supernatant to recombinant human Allergin-1L protein was analyzed by ELISA, and the binding to membrane-expressed human Allergin-1L was analyzed by CELISA. Furthermore, antibody clones that showed binding in either analysis were analyzed by CELISA for binding to CHO cells stably expressing human Allergin-1S1, human Allergin-1S2, and monkey Allergin-1. In this CELISA, clones that gave signals (OD values) 10% or higher of those obtained in the positive control wells (using previously obtained anti-human Allergin-1 antibodies) were defined as positive clones. Furthermore, surface plasmon resonance (SPR) analysis using a MASS-2 SPR device (Sierra Sensors) was used to analyze the binding of antibodies in the culture supernatant to recombinant human Allergin-1L protein, and the dissociation constant (KD value) was calculated. The analysis was performed by binding rabbit antibodies in the culture supernatant to anti-rabbit IgG antibodies immobilized on a sensor chip (SPR-2 Affinity sensor, High Capacity Amine, Bruker LabScape), followed by flowing recombinant human Allergin-1L protein as an analyte through a flow cell. From the analysis results, several antibody clones were selected, including positive clones that bound to both human Allergin-1L and monkey Allergin-1.

[0129] Example 3: DNA sequencing of candidate clones for anti-Allergin-1 antibodies RNA was extracted from each of the positive B cell clones isolated in Example 2, and cDNA was prepared by reverse transcription. PCR was performed using the prepared cDNA and primers specific to the immunoglobulin heavy and light chain variable regions. The amplified PCR products were subjected to DNA sequencing to determine the DNA sequences encoding the heavy and light chain variable region sequences of the antibody produced by the B cell clone.

[0130] Example 4: Evaluation of binding to Allergin-1-expressing cells using rabbit IgG format and screening by competitive binding assay Expression vectors encoding the heavy and light chains of the rabbit antibody whose DNA sequence was determined in Example 3 were prepared, and both expression vectors were transfected into mammalian cells to produce the antibody in the culture supernatant. The culture supernatant was collected and treated with protein A affinity chromatography to purify rabbit IgG.

[0131] The binding of purified rabbit IgG to CHO cells stably expressing human Allergin-1L, human Allergin-1S1, human Allergin-1S2, and monkey Allergin-1 was analyzed by CELISA. Specifically, CHO cells stably expressing each Allergin-1 were seeded into a 96-well plate, diluted rabbit IgG was added, and the cells were incubated at 37°C. After washing, the CHO cells were added with diluted HRP (horseradish peroxidase)-labeled goat anti-rabbit IgG antibody and incubated again at 37°C. After washing again, the CHO cells were added with a chromogenic substrate for HRP and incubated at room temperature, followed by measurement of absorbance.

[0132] A competitive binding assay was performed to evaluate cross-competitive activity of 34 anti-S1 domain antibodies confirmed to bind to human Allergin-1L and human Allergin-1S1 by CELISA, and 32 anti-S2 domain antibodies confirmed to bind to human Allergin-1L and human Allergin-1S2. Specifically, two antibodies were randomly selected from the 34 anti-S1 domain antibodies, one as a competitor antibody and the other as a detector antibody, and the binding of the detector antibody to human Allergin-1L in the presence of the competitor antibody was evaluated. First, the competitor antibody was added to CHO cells expressing human Allergin-1L and incubated on ice. An equal volume of buffer was added to samples without the competitor antibody. Next, biotin-labeled rabbit IgG was added as a detector antibody to the competitor antibody and incubated on ice. After washing, the CHO cells were incubated with APC (Allophycocyanin)-labeled streptavidin. The CHO cells were washed again, and the amount of detector antibody binding (MFI value) was measured by flow cytometry. The amount of detector antibody binding in the presence of competitor antibody was calculated, with the amount of detector antibody binding in the absence of competitor antibody set at 100. A similar analysis was performed on 32 types of anti-S2 domain antibodies.

[0133] The cross-competitive patterns of each antibody clone obtained from the competitive binding assay were analyzed, and antibodies showing similar patterns of binding to human Allergin-1L were classified into four groups sharing the same epitope (S1-A epitope, S1-B epitope, S2-C epitope, and S2-D epitope).

[0134] Example 5: Evaluation of binding and stability to Allergin-1-expressing cells using a humanized scFv format. Humanized scFvs were designed from rabbit antibodies to reduce immunogenicity in humans and stabilize the scFv structure. Specifically, the heavy and light chain complementarity-determining regions (VH-CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2, and VL-CDR3) of the rabbit antibodies selected and evaluated in Examples 3 and 4 were determined, and the DNA sequences encoding these six CDRs were then grafted onto DNA sequences encoding the framework regions of the heavy and light chains of a human antibody using techniques known to those skilled in the art. The grafted DNAs encoding the heavy and light chains were then recombined using techniques known to those skilled in the art to be expressed in the form of scFvs, and the recombined DNAs were inserted into a mammalian cell expression vector. The expression vector was transfected into mammalian cells to produce scFv in the culture supernatant, which was then collected and purified by protein L affinity chromatography to give humanized scFv.

[0135] The purified humanized scFv was analyzed for binding to CHO cells stably expressing human Allergin-1L and monkey Allergin-1 by CELISA. Specifically, the CHO cells were seeded into a 96-well plate, diluted humanized scFv was added, and the cells were incubated at 37°C. After washing, the CHO cells were added with diluted HRP-labeled Protein L and incubated again at 37°C. After washing the CHO cells again, a chromogenic substrate for HRP was added, and the cells were incubated at room temperature, after which the absorbance was measured. As a result, binding to human Allergin-1L and monkey Allergin-1 was confirmed in multiple clones.

[0136] Furthermore, the binding of each humanized scFv to Fc-fused human Allergin-1L recombinant protein was analyzed by SPR. Specifically, the Fc-fused human Allergin-1L recombinant protein was immobilized on a sensor chip, and then the humanized scFv was applied as an analyte to a flow cell. As a result, binding to human Allergin-1L was confirmed in multiple clones.

[0137] Based on the results of the above series of binding assays, 17 humanized scFvs were selected, taking into account the diversity of the epitope group, and stability was evaluated. Specifically, solutions of each selected humanized scFv in 50 mM phosphate-citrate buffer (humanized scFv: ≥ 10 mg / mL; containing 150 mM NaCl; pH 6.4) were stored at temperatures below -80°C, 4°C, and 40°C for 4 weeks. The changes in monomer ratio were evaluated by SE-HPLC after 1, 7, 14, 21, 28, and 35 days of storage, and the changes in protein concentration were measured by absorbance at 280 nm. To assess the effect of freeze-thaw cycles on stability, the changes in monomer ratio were also evaluated by SE-HPLC after up to four freeze-thaw cycles. Furthermore, thermal stability was evaluated using differential scanning fluorimetry (DSF).

[0138] Taking into consideration the binding to human Allergin-1L and monkey Allergin-1 as determined by CELISA and SPR, as well as the diversity of epitope groups, eight humanized scFvs shown in Table 1 were selected from among those with a small decrease in monomer ratio in a 14-day stability evaluation as humanized scFvs having VH and VL to be used in the subsequent design of anti-Allergin-1 bispecific antibodies.

[0139]

[0140] Two different types of humanized scFvs were randomly selected from the eight types of humanized scFvs selected in Example 5, and anti-Allergin-1 bispecific antibodies were designed, each containing two binding sites derived from the two humanized scFvs. Specifically, two types of humanized scFvs that did not compete with each other (confirmed by the competitive binding assay shown in Example 4) were selected from the eight types of humanized scFvs selected, and the VH and VL of one humanized scFv were designed to form the Fab region of the scFv, with the other humanized scFv linked to the C-terminus of the heavy chain or light chain, forming a so-called Morrison-form antibody. When linking the humanized scFv to the C-terminus of the heavy chain or light chain, the linker sequence Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 73) was used. The DNA encoding the heavy and light chains constituting the anti-Allergin-1 bispecific antibody and their expression vectors were constructed according to the design using genetic recombination techniques known to those skilled in the art. pcDNA3.4 (Thermo Fisher Scientific) was used as the expression vector. The constructed expression vectors were then simultaneously transfected into HEK293 cells, and the antibody was produced in the culture supernatant. The culture supernatant was collected and subjected to protein A affinity chromatography to purify the desired anti-Allergin-1 bispecific antibody. The structure of some of the constructed anti-Allergin-1 bispecific antibodies is shown in Table 2.

[0141]

[0142] Example 7: Evaluation of the degree of shielding by binding of anti-Allergin-1 bispecific antibodies to Allergin-1 For receptors with unknown ligands, antibodies that cover a wider range of the extracellular portion are preferred. Therefore, the degree of shielding by binding of the anti-Allergin-1 bispecific antibodies prepared in Example 6 to Allergin-1 was evaluated.

[0143] First, each anti-Allergin-1 bispecific antibody (antibody clones #1 to #20 in the table in Figure 16 ) prepared according to the same procedure as in Example 6 was added to a human Allergin-1L-expressing CHO cell line, followed by incubation on ice. Furthermore, each biotin-labeled anti-Allergin-1 antibody (rabbit IgG obtained in Example 4: each antibody listed in the vertical column labeled "Rabbit IgG ID No." in the table in Figure 16 ) was added, followed by incubation on ice again. After washing the CHO cells, APC-labeled streptavidin was added, followed by incubation on ice again. After washing the CHO cells again, the amount of binding of the biotin-labeled anti-Allergin-1 antibody was assessed using a flow cytometer. The results are shown in Figure 16 .

[0144] It was confirmed that each anti-Allergin-1 bispecific antibody inhibits the binding of many anti-Allergin-1 antibodies to Allergin-1L and widely shields the surface of Allergin-1L.

[0145] Example 8: Construction of mutants of anti-Allergin-1 bispecific antibodies Mutants that retained the antigen recognition ability of anti-Allergin-1 bispecific antibodies were constructed using site-directed mutagenesis techniques known to those skilled in the art.

[0146] The anti-Allergin-1 bispecific antibody #15_h clone produced in Example 6 was subjected to site-directed mutagenesis so as to substitute the aspartic acid at position 461 with alanine, the serine at position 474 with arginine, the aspartic acid at position 532 with glutamine, the threonine at position 565 with cysteine, and the glycine at position 637 with cysteine ​​in SEQ ID NO: 77, which represents the amino acid sequence of its heavy chain, to produce the anti-Allergin-1 bispecific antibody #15_h_opt clone having a heavy chain consisting of the amino acid sequence of SEQ ID NO: 79.

[0147] Furthermore, the #15_h clone was separately subjected to site-directed mutagenesis to replace aspartic acid at position 461 with alanine, serine at position 474 with arginine, and aspartic acid at position 532 with glutamine in SEQ ID NO: 77, thereby generating the anti-Allergin-1 bispecific antibody #15_h_optv2 clone having a heavy chain consisting of the amino acid sequence of SEQ ID NO: 98.

[0148] On the other hand, the anti-Allergin-1 bispecific antibody #15p_h clone was prepared by swapping the two antigen recognition sites (on the Fab and scFv sides) of the #15_h clone. (i) Site-directed mutagenesis was performed on the heavy chain amino acid sequence of SEQ ID NO: 83, substituting threonine at position 562 with cysteine ​​and glycine at position 634 with cysteine, to produce a heavy chain having the amino acid sequence of SEQ ID NO: 84, and (ii) Site-directed mutagenesis was performed on the light chain amino acid sequence of SEQ ID NO: 82, substituting aspartic acid at position 1 with alanine, serine at position 14 with arginine, and aspartic acid at position 72 with glutamine, to produce the anti-Allergin-1 bispecific antibody #15p_h_opt clone (see Table 3).

[0149]

[0150] Flow cytometry revealed that the binding activity of the mutants created here to Allergin-1L (EC ５０ As shown in Table 4, the binding activity of each of the clones was equivalent to that of the original #15_h clone.

[0151]

[0152] Example 9: Evaluation of binding activity of anti-Allergin-1 bispecific antibodies The contribution of each binding site of the anti-Allergin-1 bispecific antibodies prepared in Example 8 to binding to Allergin-1L was evaluated.

[0153] First, CHO cells expressing human Allergin-1L were incubated with either the rabbit anti-Allergin-1S1 IgG antibody #85 clone (described in Example 4), which has the same CDR as the scFv of the anti-Allergin-1 bispecific antibody #15_h_opt clone, or the rabbit anti-Allergin-1S2 IgG antibody #50 clone (described in Example 4), which has the same CDR as the Fab of the anti-Allergin-1 bispecific antibody #15_h_opt clone, or both. After washing, the CHO cells were incubated with 340 ng / mL of the #15_h_opt clone. After washing again, the CHO cells were incubated with an HRP-labeled anti-human Fc antibody. After further washing, the CHO cells were incubated with a chromogenic substrate for HRP, and the absorbance was measured. The results are shown in Figure 17.

[0154] Addition of either clone #85 or clone #50 alone did not completely inhibit the binding of clone #15_h_opt to Allergin-1L, but simultaneous addition of both clones completely inhibited the binding, confirming that each binding site of the anti-Allergin-1 bispecific antibody clone #15_h_opt contributes to the binding to Allergin-1L.

[0155] Example 10: Evaluation of binding characteristics of anti-Allergin-1 bispecific antibodies A competitive binding assay was performed to evaluate the effect of the anti-Allergin-1 bispecific antibodies prepared in Example 8 on the binding of ODN M362, known as a TLR9 ligand, to Allergin-1.

[0156] First, 6xHis tagged human Allergin-1L protein was prepared using a method known to those skilled in the art and immobilized on a Series S Sensor Chip CM5 (Cytiva, model number 29149603) using a His Capture Kit (Cytiva, model number 28995056). The anti-Allergin-1 bispecific antibody #15_h_opt clone, rabbit anti-Allergin-1S1 IgG antibody #85 clone, and rabbit anti-Allergin-1S2 IgG antibody #50 clone were each added to the Series S Sensor Chip CM5. The binding of ODN M362 (InvivoGen, model number tlrl-m362-1) to the immobilized 6xHis tagged human Allergin-1L protein was then evaluated using Biacore (Biacore T200, Cytiva). The results are shown in Figure 18.

[0157] The #15_h_opt clone and the #85 clone inhibited the binding of ODN M362 to the 6xHis tag-fused human Allergin-1L protein, with the #15_h_opt clone exhibiting stronger inhibition.

[0158] Example 11: Evaluation of in vivo activity of anti-Allergin-1 bispecific antibodies in a mouse cancer-bearing model The in vivo activity of the anti-Allergin-1 bispecific antibodies prepared in Example 6 was evaluated in a cancer-bearing model using human Allergin-1L knock-in C57BL / 6 mice, which were generated by replacing the Allergin-1 gene with the human Allergin-1L gene using techniques known to those skilled in the art.

[0159] The human Allergin-1L knock-in C57BL / 6 mice were injected with 2 x 10 ５Mouse colon cancer cell line MC38 was subcutaneously implanted, and the anti-mouse PD-1 antibody 4H2 (see WO2006 / 121168) was intraperitoneally administered on the day of implantation, 6 days, 12 days, and 17 days after implantation. Meanwhile, anti-Allergin-1 bispecific antibodies (#11_L, #13_L, #15_L, #16_L, and #15_h clones) were intraperitoneally administered together with the anti-mouse PD-1 antibody 4H2 according to the above-mentioned administration schedule on the day of implantation, and 4, 7, 10, 14, 18, and 21 days after implantation. Starting 7 days after implantation, the long and short diameters of the tumor were measured twice a week, and tumor volume was calculated using the formula (long diameter) × (short diameter) 2 × 0.5. For mice with multiple tumors, the tumor volumes were calculated individually, and the sum of these values ​​was used as the tumor volume for that mouse. The results are shown in Figure 19.

[0160] All of the anti-Allergin-1 bispecific antibodies evaluated significantly enhanced the antitumor effect of the anti-mouse PD-1 antibody 4H2 when administered in combination with the antibody.

[0161] Example 12: Evaluation of in vivo activity of anti-Allergin-1 bispecific antibodies in a mouse cancer-bearing model The in vivo activity of the anti-Allergin-1 bispecific antibodies prepared in Example 6 was evaluated in the human Allergin-1L knock-in C57BL / 6 mouse cancer-bearing model described in Example 11.

[0162] The human Allergin-1L knock-in C57BL / 6 mice were injected with 2 x 10 ５ Mouse lung cancer cell line 3LL was subcutaneously implanted, and the anti-mouse PD-1 antibody 4H2 was administered intraperitoneally 6, 12, and 18 days after implantation. Meanwhile, the anti-Allergin-1 bispecific antibody #15_h clone was administered intraperitoneally along with the anti-mouse PD-1 antibody 4H2 according to the above schedule on the day of implantation, and 4, 7, 11, 14, 18, and 21 days after implantation. Starting 7 days after implantation, the long and short diameters of the tumor were measured twice a week, and tumor volume was calculated using the formula (long diameter) × (short diameter) 2 × 0.5. For mice with multiple tumors, the tumor volumes were calculated individually, and the sum of these values ​​was used as the tumor volume for that mouse. The results are shown in Figure 20.

[0163] When used in combination with the anti-mouse PD-1 antibody 4H2, the #15_h clone enhanced the antitumor effect, which was hardly observed when 4H2 was administered alone.

[0164] Example 13: Evaluation of in vivo activity of anti-Allergin-1 bispecific antibodies in a mouse cancer-bearing model The in vivo activity of the anti-Allergin-1 bispecific antibodies prepared in Example 8 was evaluated in the human Allergin-1L knock-in C57BL / 6 mouse cancer-bearing model described in Example 11.

[0165] The human Allergin-1L knock-in C57BL / 6 mice were injected with 6 x 10 ４ MC38 mice overexpressing human PD-L1 were subcutaneously implanted, and the anti-Allergin-1 bispecific antibody #15_h_opt clone was administered intraperitoneally on the day of implantation, 7 days after implantation, and 14 days after implantation. Starting 7 days after implantation, the long and short diameters of the tumors were measured twice a week, and tumor volume was calculated using the formula (long diameter) × (short diameter) 2 × 0.5. For mice with multiple tumors, the tumor volumes of each mouse were calculated, and the sum of these values ​​was used as the tumor volume for that mouse. The results are shown in Figure 21.

[0166] The #15_h_opt clone also showed antitumor effects when administered alone.

[0167] Example 14: Evaluation of in vivo effects of anti-Allergin-1 bispecific antibodies on tumor-infiltrating CD45-positive cells in a mouse cancer-bearing model In the human Allergin-1L knock-in C57BL / 6 mouse cancer-bearing model of Example 11, the in vivo effects of the anti-Allergin-1 bispecific antibodies prepared in Examples 6 and 8 on tumor-infiltrating CD45-positive cells were evaluated.

[0168] The human Allergin-1L knock-in C57BL / 6 mice were injected with 2 x 10 ５Mouse colon cancer cell line MC38 cells were subcutaneously implanted, and the anti-Allergin-1 bispecific antibodies #15_h, #15_h_opt, and #15p_h_opt clones were intraperitoneally administered on the day of implantation, 4 days after implantation, and 7 days after implantation, along with the anti-mouse PD-1 antibody 4H2 (administered on the day of implantation and 6 days after implantation, respectively). Tumor tissue was excised 11 days after implantation, and CD45-positive cells were purified. mRNA was extracted from the purified cells, cDNA was synthesized by reverse transcription, and expression of various genes was analyzed by quantitative PCR. The results are shown in Table 5.

[0169]

[0170] The numbers in the table represent the expression ratios of various genes relative to the expression levels in the group administered with the anti-mouse PD-1 antibody 4H2 alone.

[0171] When used in combination with the anti-mouse PD-1 antibody 4H2, each of these anti-Allergin-1 bispecific antibody clones induced the expression of anti-tumor immune-related genes Gzmb, Prf1, Ifng, Ifnb1, Ifna2, Ifna4, and Ifna6.

[0172] Example 15: Evaluation of the in vivo effect of anti-Allergin-1 bispecific antibodies on tumor-infiltrating CD45-positive cells in a mouse cancer-bearing model The in vivo effect of the anti-Allergin-1 bispecific antibodies produced in Example 8 on tumor-infiltrating CD45-positive cells was evaluated in the human Allergin-1L knock-in C57BL / 6 mouse cancer-bearing model of Example 11.

[0173] The human Allergin-1L knock-in C57BL / 6 mice were injected with 6 × 10 ４ MC38 tumors expressing human PD-L1 were subcutaneously implanted, and the anti-Allergin-1 bispecific antibody #15_h_opt clone was administered intraperitoneally on the day of implantation, 8 days after implantation, and 14 days after implantation. Tumor tissue was excised 10 or 15 days after implantation, and CD45-positive cells were purified. mRNA was extracted from the purified cells, and cDNA was synthesized by reverse transcription. Expression of various genes was then analyzed by quantitative PCR. The results are shown in Table 6.

[0174]

[0175] The numbers in the table represent the ratio of the expression level of each gene to the expression level of the control group.

[0176] Even when administered alone, the #15_h_opt clone induced the expression of anti-tumor immune-related genes Gzmb, Prf1, Ifng, Ifnb1, and Ifna4.

[0177] Example 16: Identification of the epitope of the anti-Allergin-1 bispecific antibody #15_h_opt clone. We prepared a 6xHis-tagged human Allergin-1L (C38S) protein (deleting amino acids 20-21 and 217 and beyond), an Fv-clasp protein (S1-Fv-clasp) with the same CDRs as the scFv of #15_h_opt clone (see Structure, 2017, Vol. 25, No. 10, pp. 1611-1622), and an Fv-clasp protein (S2-Fv-clasp) with the same CDRs as the Fab of #15_h_opt clone, and crystals of the ternary complex were obtained. Crystals were frozen using the free mounting system (FMS) method, and X-ray diffraction data were acquired. The structure was refined using data corrected by STARANISO, and the structure was determined at 2.62 Å resolution.

[0178] The determined structure revealed that the S1-A epitope is composed of at least one or more of the following residues of Allergin-1: phenylalanine at position 70, arginine at position 71, arginine at position 72, lysine at position 73, histidine at position 75, leucine at position 76, histidine at position 96, glutamic acid at position 97, lysine at position 102, lysine at position 104, and tyrosine at position 113; and the S2-D epitope is composed of at least one or more of the following residues of Allergin-1: isoleucine at position 132, methionine at position 133, valine at position 134, isoleucine at position 135, glutamine at position 136, threonine at position 137, glutamic acid at position 138, histidine at position 146, and proline at position 215.

[0179] Example 17: Evaluation of the in vitro effect of anti-Allergin-1 bispecific antibodies on cytokine release from human peripheral blood mononuclear cells An experiment was carried out in which the anti-Allergin-1 bispecific antibodies prepared in Example 8 were added to human peripheral blood mononuclear cells (hereinafter referred to as "human PBMCs").

[0180] Human PBMCs (Lonza, product number CC-2702) were cultured for 24 hours after the addition of the anti-Allergin-1 bispecific antibody #15_h_opt clone at final concentrations of 1 ng / mL to 30 μg / mL and the positive control anti-human CD3 antibody at 10 μg / mL. After 24 hours, the levels of IFN-γ, TNF, IL-10, IL-6, IL-4, and IL-2 in the culture supernatant were quantified using a Cytometric Bead Array Human Th1 / Th2 Cytokine Kit II (BD Biosciences, product number 551809). The results are shown in Figure 22. Cytokine production levels (pg / mL) are expressed as mean ± standard error (N = 3).

[0181] While the anti-CD3 antibody induced the production of all cytokines, the #15_h_opt clone did not significantly induce any of the cytokines.

[0182] The anti-Allergin-1 bispecific antibodies of the present invention are useful for preventing cancer, inhibiting the progression of symptoms, inhibiting recurrence, or treating cancer.

Claims

[Claim 1] A bispecific antibody against Allergin-1 or an antibody fragment thereof, comprising two different antigen-binding sites selected from the S1-A binding site, S1-B binding site, S2-C binding site, and S2-D binding site, The S1-A binding site is (A) (a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 25, (b) VH-CDR2 and the amino acid sequence of SEQ ID NO: 26 (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 27, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 49, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 50 and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 51, (B) (a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 28, (b) VH-CDR2 and the amino acid sequence of SEQ ID NO: 29 (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 30, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 52, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 53 and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 54, (C)(a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 31, (b) VH-CDR2 and the amino acid sequence of SEQ ID NO: 32 (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 33, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 55, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 56 and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 57, and (D) (a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 34, (b) VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 35 and (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 36, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 58, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 59 and (f) Any one antigen-binding site selected from the antigen-binding sites consisting of VL having VL-CDR3, which is the amino acid sequence of SEQ ID NO: 60, The S1-B binding site is (E)(a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 37, (b) VH-CDR2 and the amino acid sequence of SEQ ID NO: 38 (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 39, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 61, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 62 and (f) Antigen binding site consisting of VL having VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 63 or (F)(a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 40, (b) VH-CDR2 and the amino acid sequence of SEQ ID NO: 41 (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 42, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 64, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 65 and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 66, The S2-C binding site is (a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 43, (b) VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 44 and (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 45, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 67, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 68 and (f) An antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 69, The S2-D binding site is (a) VH-CDR1 consisting of the amino acid sequence of SEQ ID NO: 46, (b) VH-CDR2 consisting of the amino acid sequence of SEQ ID NO: 47 and (c) VH having VH-CDR3 consisting of the amino acid sequence of SEQ ID NO: 48, and (d) VL-CDR1 consisting of the amino acid sequence of SEQ ID NO: 70, (e) VL-CDR2 consisting of the amino acid sequence of SEQ ID NO: 71 and (f) The bispecific antibody or antibody fragment, which has an antigen-binding site consisting of a VL having a VL-CDR3 consisting of the amino acid sequence of SEQ ID NO: 72.