Antigen-binding polypeptides, antigen-binding polypeptide complexes, and methods of using same

Abstract

Disclosed are antigen-binding polypeptides and antigen-binding polypeptide complexes (e.g., antibodies and antigen-binding fragments thereof) having certain structural characteristics. Also disclosed are polynucleotides and vectors encoding such polypeptides and polypeptide complexes, host cells, chimeric antigen receptors (CARs), immune cells, pharmaceutical compositions, and kits containing such polypeptides and polypeptide complexes, and methods of using such polypeptides and polypeptide complexes.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 249,833, filed September 29, 2021, U.S. Provisional Application No. 63 / 249,794, filed September 29, 2021, U.S. Provisional Application No. 63 / 249,919, filed September 29, 2021, U.S. Provisional Application No. 63 / 249,722, filed September 29, 2021, U.S. Provisional Application No. 63 / 291,305, filed December 17, 2021, and U.S. Provisional Application No. 63 / 292,382, filed December 21, 2021, all of which are incorporated by reference in their entirety.

[0002] Reference to an electronically submitted sequence listing The contents of the electronically submitted sequence listing (Name: 4850_003PC01_SeqListing_ST26; Size: 1,196,609; Creation Date: September 26, 2022) are incorporated herein by reference in their entirety.

[0003] Field The present disclosure relates to antigen-binding polypeptides and antigen-binding polypeptide complexes (e.g., antibodies and antigen-binding fragments thereof) having certain structural characteristics. The disclosure also relates to polynucleotides and vectors encoding such polypeptides and polypeptide complexes, host cells containing such polypeptides and polypeptide complexes, chimeric antigen receptors (CARs), immune cells, pharmaceutical compositions, and kits, as well as methods of using such polypeptides and polypeptide complexes. [Background technology]

[0004] background Immunotherapy is the treatment of diseases by activating or suppressing the immune system. In recent years, immunotherapy has become of great interest to researchers and clinicians, especially in its potential to treat cancer and infectious diseases. Therapeutic antibodies are an important type of immunotherapy. Therapeutic antibodies can be monospecific, meaning they have specificity for one antigen or epitope. Therapeutic antibodies have also been engineered to have specificity for two different antigens or epitopes (i.e., bispecific antibodies) or multiple different antigens or epitopes (trispecific antibodies, tetraspecific antibodies, etc.). In addition, monospecific antibodies, bispecific antibodies, and multispecific antibodies have been combined to form multitargeting strategies for treating complex human diseases such as cancer and infectious diseases.

[0005] However, the development of therapeutic antibodies can be challenging, especially in manufacturing and later stages of development. For example, the production of bispecific or multispecific antibodies often requires multiple genes or plasmids for cell line development. These multiple genes or plasmids must be delivered to the same cell to produce the correct molecule. Furthermore, bispecific and multispecific antibodies may have mismatched heavy and light chains, which can reduce product yields, increase cell line colony screening workload, and result in product heterogeneity.

[0006] A need exists for multispecific and multifunctional antigen-binding polypeptides and antigen-binding polypeptide complexes that can bind to specific combinations of target molecules for selectivity or amplification / neutralization, bring together two or more cell types, bring together targets to deliver activating signals, modify the disease microenvironment, and enhance binding affinity for improved efficacy. The present invention fulfills this unmet need.

[0007] Additionally, human immunodeficiency virus (HIV) imposes a major infectious disease burden worldwide with enormous medical and economic impact. Globally, approximately 38 million people are infected with HIV, and more than 30 million people suffer from acquired immunodeficiency syndrome (AIDS), a chronic condition of weakened immune system caused by HIV infection. "Global Health Sector Strategy On HIV-2016-2021 - Towards Ending AIDS," World Health Organization, June 2016 (Non-Patent Document 1). There are two major forms of HIV: HIV-1 and HIV-2. HIV-1 is the more common form worldwide, while HIV-2 is less pathogenic and is primarily limited to West Africa.

[0008] The major structural proteins of HIV are Gag, Pol, and Env. Gag (group-specific antigen) is a structural protein of the viral core. Pol is a polyprotein containing enzymes important for viral replication: protease (PR), reverse transcriptase (RT), and integrase (IN). Env (envelope) encodes the glycoprotein that forms the outer envelope of the virus. Env is synthesized as a precursor glycoprotein, gp160, which is then processed into gp120 and gp41. Env interacts with the primary receptor, CD4, and co-receptors (such as the chemokine receptor CCR5) to fuse the viral and target cell membranes.

[0009] The genetic heterogeneity and glycan shield of Env resist the development of natural immunity against HIV, posing a challenge to conventional vaccine development and prompting the search for alternative approaches to HIV prevention, one of the highest global health priorities.

[0010] Despite the availability of a significant selection of anti-HIV / AIDS drugs, HIV patients still face the daily challenges of taking multiple medications in rigorous regimens. Inevitably, most patients endure the consequences of the emergence of drug-resistant viral variants and develop other health problems, such as cardiovascular disease, kidney disease, diabetes, bone disease, liver disease, and cognitive impairment, from the toxicity of long-term anti-HIV medications. Alternative treatment options are urgently needed for HIV / AIDS patients.

[0011] Broadly neutralizing HIV-1 antibodies (bnAbs) are antibodies that neutralize multiple HIV-1 virus strains. bnAbs target conserved epitopes on the virus, meaning that the targeted epitopes are more likely to remain even when the virus mutates. Therefore, bnAbs have recently been investigated for the treatment and prevention of HIV / AIDS. Human clinical studies have revealed two key factors for the efficacy of bnAbs. First, there is a need to exceed the minimum effective dose or trough level of circulating bnAbs to prevent infection. Second, there is a need to prevent the emergence of viral escape through resistance mutations.

[0012] Early human clinical studies using bnAbs demonstrated the feasibility and safety of this approach, with transient reductions in viral load and acceptable tolerability and immunogenicity. Burton et al., Annu. Rev. Immunol. 34:635-659 (2016) (Non-Patent Document 2), Mascola et al., Immunol. Rev. 254:225-244 (2013) (Non-Patent Document 3), Wu et al., Science. 329:856-861 (2010) (Non-Patent Document 4). However, resistant HIV strains rapidly emerged after treatment with individual bnAbs in vitro and in vivo. More recently, a phase II clinical trial using the VRC01 bnAb emphasized the importance of maintaining adequate circulating antibody levels to reduce acquisition rates, suggesting that combination antibody therapies that improve efficacy and minimize escape mutations will be required for effective prevention. Corey et al., N. Engl. J. Med. 384:1003-1014 (2021) (Non-patent Document 5).

[0013] Multispecific antibodies address the limitations of bnAbs by providing a single antibody type that recognizes multiple independent binding sites on the HIV-1 envelope protein. Xu et al., Science. 358(6359):85-90(2017) (Non-Patent Document 6). Treatment with multispecific antibodies also ensures that independent binding specificities maintain the same pharmacokinetics, while treatment with multiple single-targeting antibodies results in different antibody half-lives that decline at different rates. Furthermore, multispecific antibodies simplify manufacturing and administrative procedures by using a single product for clinical development instead of a combination of multiple products.

[0014] Thus, multispecific anti-HIV antibodies provide an important technology platform for developing neutralizing antibody-based therapeutics for treating HIV / AIDS, offering a class of drugs with low long-term toxicity and significantly less frequent treatment regimens. Multispecific antibodies also complement existing drugs by using an HIV target that is completely different from current standard-of-care HIV / AIDS drugs, offering patients an alternative for disease control and health management. Multispecific antibodies may also offer a meaningful approach to HIV prevention, where no effective HIV vaccine currently exists.

[0015] In addition, the development of therapeutic antibodies can be challenging, especially in manufacturing and later stages of development. For example, the production of multispecific antibodies often requires multiple genes or plasmids for cell line development. These multiple genes or plasmids must be delivered to the same cell to produce the correct molecule. Furthermore, multispecific antibodies may have mismatched heavy and light chains, which can reduce product yields, increase cell line colony screening workload, and result in product heterogeneity.

[0016] Thus, there is a need for multispecific and multifunctional antibodies, antigen-binding polypeptides, and antigen-binding polypeptide complexes that can bind to HIV proteins for selectivity or amplification / neutralization, bring together two or more cell types, bring together targets to deliver activation signals, modify the HIV microenvironment, and enhance binding affinity to improve efficacy. The present invention fulfills this unmet need. [Prior art documents] [Non-patent literature]

[0017] [Non-Patent Document 1] “Global Health Sector Strategy On HIV-2016-2021-Towards Ending AIDS,” World Health Organization, June 2016 [Non-patent document 2] Burton et al.,Annu.Rev.Immunol.34:635-659(2016) [Non-patent document 3] Mascola et al.,Immunol.Rev.254:225-244(2013) [Non-patent document 4] Wu et al.,Science.329:856-861(2010) [Non-patent document 5] Corey et al.,N.Engl.J.Med.384:1003-1014(2021) [Non-patent document 6] Xu et al.,Science.358(6359):85-90(2017) Summary of the Invention

[0018] Quick Overview Provided herein are antigen-binding polypeptides having a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, and L1, L2, and L3 are amino acid linkers.

[0019] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-VH2-VL2-VL1, VL1-VH2-VL2-VL2-L3-VL1, or VL1-L1-VH2-L2-VL2-L3-VL1. and VH1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, and L1, L2, and L3 are amino acid linkers.

[0020] Provided herein are antigen-binding polypeptides having a structure represented by VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, wherein VL1 is is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, and L4 are amino acid linkers.

[0021] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, and the second polypeptide is VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L 1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, and L4 are amino acid linkers.

[0022] Provided herein are VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-CL, VH1-L1-VH2-L2- VL2-L3-VL1-L4-CH1-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-CH1, VH1-L1-VH2-L2-VL2-L3- VL1-L4-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L 4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0023] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1- VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL 1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1- and the second polypeptide has a structure represented by VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, and the second polypeptide is VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-C H1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1- L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL 1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1,or an antigen-binding polypeptide complex having a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0024] Provided herein are VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L 4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH 2-L3-VH1-L4-CL-L5-CH1-Fc, or by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0025] Provided herein is an antigen-binding polypeptide complex comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2- VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-F c, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1- CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL 1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2- VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc,and VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, and VH1 is a first immunoglobulin light chain variable region. and L1, L2, L3, L4, and L5 are amino acid linkers.

[0026] Provided herein is an antigen binding polypeptide complex containing the 1st polypeptide and the 2nd polypeptide VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1- L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-V H2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2 -VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-V L1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-Fc CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L 3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL It has the structure shown by 2-L3-VL1-L4-CL-L5-CH1-Fc. VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-Fc L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL L1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、 VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-C H1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2 -VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL and VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, and VH1 is a first immunoglobulin heavy chain variable region. VH1 is a second immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0027] Provided herein are VL1-VL2-VH2-VH1-Fc-Fc, VH1-VH2-VL2-VL1-Fc-Fc, VL1-L1-VL2-L2 -VH2-L3-VH1-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc-Fc, VL1-L1-VL2-L2-VH2-L3- a polypeptide having a structure represented by VH1-L4-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-L5-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc-L5-Fc wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0028] Provided herein are VL1-VL2-VH2-VH1-CH3, VH1-VH2-VL2-VL1-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-CH3, VL1-L1-VL2-L2-VH2-L3-VL1-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-L4 -CH3, VL1-VL2-VH2-VH1-CH3-CH3, VH1-VH2-VL2-VL1-CH3-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-CH3-CH3 , VH1-L1-VH2-L2-VL2-L3-VL1-CH3-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH3-CH3, VH1-L1-VH2-L2-V an antigen-binding polypeptide or antigen-binding polypeptide complex comprising a polypeptide having the structure represented by L2-L3-VL1-L4-CH3-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH3-L5-CH3, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH3-L5-CH3, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH3 is immunoglobulin heavy chain constant region 3, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0029] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, and the second polypeptide has a structure represented by VL3-VL4-VH4-VH3, VH3-VH4-VL4-VL3, VL3-L4-VL4-L5-VH4-L6-VH3, or VH3-L4-VH4-L5-VL4-L6-VL3. wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, VL4 is a fourth immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, VH3 is a third immunoglobulin heavy chain variable region, VH4 is a fourth immunoglobulin heavy chain variable region, and L1, L2, L3, L4, L5, and L6 are amino acid linkers.

[0030] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- and the second polypeptide has a structure represented by VL3-VL4-VH4-VH3-Fc, VH3-VH4-VL4-VL3-Fc, VL3-L5-VL4-L6-VH4-L7-VH3-Fc, VH3-L5-VH4-L6-VL4-L7-VL3-Fc, VL3-L5-VL4-L6-VH4-L7-VH3-Fc, VL3-L5-VL4-L6-VH4-L7-VH3-L8-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, has a structure represented by VH3-L5-VH4-L6-VL4-L7-VL3-L8-Fc, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, and VL4 is a fourth immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, and VH2 is a second immunoglobulin light chain variable region. VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, VH3 is a third immunoglobulin heavy chain variable region, VH4 is a fourth immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, L5, L6, L7, and L8 are amino acid linkers.

[0031] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4 -CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, and the second polypeptide has a structure represented by VL3-VL4-VH4-VH3-CH1, VH3-VH4-VL4-VL3-CH1, VL3-VL4-VH4-VH3-CL, VH3-VH4-VL4-VL3-CL, VL3-VL4-VH4-VH3-CH1-CL, VH3-VH4-VL4-VL3-CH1-CL, VH3-VL4-VH4-VL3-CH1-CL, VH3-VL4-VH4-VL3-CH1-CL, VL3-VL4-VH4-VH3-CL-CH1, VH3-VH4-VL4-VL3-CL-CH1, VH3-VH4-VL4-VL3-CL-CH1, VL3- L6-VL4-L7-VH4-L8-VH3-L9-CH1, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL, VH3-L6-VH4-L7-VL4-L8-VL3-L9- CL, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1-L10-CL, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1-L10-CL, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL-L10-CH1,or has a structure represented by VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-L10-CH1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, VL4 is a fourth immunoglobulin light chain variable region, and VH1 is a first immunoglobulin heavy chain variable region; an antigen-binding polypeptide complex, wherein VH2 is a second immunoglobulin heavy chain variable region, VH3 is a third immunoglobulin heavy chain variable region, VH4 is a fourth immunoglobulin heavy chain variable region, CH1 is an immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, L5, L6, L7, L8, L9, and L10 are amino acid linkers.

[0032] Provided herein are antigen-binding polypeptide complexes comprising a first polypeptide and a second polypeptide, wherein the first polypeptide is selected from the group consisting of VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2 -VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL- Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL and the second polypeptide has a structure represented by VL3-VL4-VH4-VH3-CH1-Fc, VH3-VH4-VL4-VL3-CH1-Fc, VH3-VH4-VL4-VH3-CL-Fc, VH3-VL4-VH4-VH3-CL-Fc, VH3-VL4-VH4-VH3-CL-Fc, VH3-VL4-VH4-VH3-CH1-CL-Fc, VH3-VL4-VH4-VL3-C H1-CL-Fc, VL3-VL4-VH4-VH3-CL-CH1-Fc, VH3-VH4-VL4-VL3-CL-CH1-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1-Fc, VH3-L6-VH4-L7-VL4-L8-VL3 -L9-CH1-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL-Fc, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1-L10-CL-Fc,VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1-L10-CL-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL-L10-CH1-Fc, or VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-L10-CH1-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, VL4 is a fourth immunoglobulin light chain variable region, and VH1 is a first immunoglobulin heavy chain variable region. VH1 is a second immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, VH3 is a third immunoglobulin heavy chain variable region, VH4 is a fourth immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, L5, L6, L7, L8, L9, and L10 are amino acid linkers.

[0033] Provided herein is an antigen binding polypeptide complex containing the 1st polypeptide and the 2nd polypeptide VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH 2-VL2-VL1-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VH1-CL L2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-Fc CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-C L-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-Fc VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1- L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L 4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、またはVH1-L1-VH2-L2-VL2- L3-VL1-L4-CL-L5-CH1-Fc 、VH3-VH4-VL4-VL3、VL3-VL4-VH4-VH3-Fc、VH3-VH4-VL4-VL3-Fc、VL3-VL4 -VH4-VH3-CH1、VH3-VH4-VL4-VL3-CH1、VL3-VL4-VH4-VH3-CL、VH3-VH4-VL 4-VL3-CL、VL3-VL4-VH4-VH3-CH1-CL、VH3-VH4-VL4-VL3-CH1-CL、VL3-VL4 -VH4-VH3-CL-CH1、VH3-VH4-VL4-VL3-CL-CH1、VL3-VL4-VH4-VH3-CH1-Fc、 VH3-VH4-VL4-VL3-CH1-Fc、VL3-VL4-VH4-VH3-CL-Fc、VH3-VH4-VL4-VL3-C L-Fc, VL3-VL4-VH4-VH3-CH1-CL-Fc, VH3-VH4-VL4-VL3-CH1-CL-Fc, VL3-VL4-VH4-VH3-CL-CH1-Fc, VH3-VH4-VL4-VL3-CL-CH1-Fc, VL3-L6-VL4-L7-V H4-L8-VH3, VH3-L6-VH4-L7-VL4-L8-VL3, VL3-L6-VL4-L7-VH4-L8-VH3-Fc, VH3-L6-VH4-L7-VL4-L8-VL3-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-Fc, VH 3-L6-VH4-L7-VL4-L8-VL3-L9-Fc、VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1、VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1、VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL、VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1-L10-CL, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1-L10-CL, VL 3-L6-VL4-L7-VH4-L8-VH3-L9-CL-L10-CH1, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-L10-CH1, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CH1-Fc, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL-Fc, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-Fc, VL3-L6-V L4-L7-VH4-L8-VH3-L9-CH1-L10-CL-Fc, VH3-L6-VH4-L7-VL4-L8-VL3-L9-CH1-L10-CL-Fc, VL3-L6-VL4-L7-VH4-L8-VH3-L9-CL-L10-C H1-Fc, or VH3-L6-VH4-L7-VL4-L8-VL3-L9-CL-L10-CH1-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, and VL4 is a fourth immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, and VH3 is a fourth immunoglobulin light chain variable region. VH3 is an immunoglobulin heavy chain variable region, VH4 is a fourth immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, L5, L6, L7, L8, L9, and L10 are amino acid linkers.

[0034] Also provided herein are antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to viral peptides or HIV proteins.

[0035] Also provided herein is an antibody or antigen-binding fragment thereof comprising an antigen-binding polypeptide or antigen-binding polypeptide complex described herein.

[0036] Provided herein are polypeptides having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 32-43, 62-79, 130-138, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, and 679. Also provided herein are polypeptides having at least 90% identity, at least 95% identity, or 100% identity to the amino acid sequence of SEQ ID NO: 32 or 33 that does not contain the eight histidine residues at the C-terminus. Also provided herein are polypeptides encoded by polynucleotides having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 44-55, 80-97, 139-147, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, and 680.

[0037] Provided herein are polynucleotides encoding the antigen-binding polypeptides or antigen-binding polypeptide complexes described herein. Also provided herein are polynucleotides having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 44-55, 80-97, 139-147, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, and 680. Also provided herein are polynucleotides that encode a polypeptide having at least 90%, at least 95%, or 100% identity to any one of SEQ ID NOs: 32-43, 62-79, 130-138, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, and 679, or polynucleotides that encode a polypeptide having at least 90%, at least 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 32 or 33 that does not contain the eight histidine residues at the C-terminus.

[0038] Provided herein are vectors that include the polynucleotides described herein.

[0039] Provided herein are host cells containing the polynucleotides or vectors described herein.

[0040] Provided herein is a chimeric antigen receptor (CAR) comprising an antigen-binding polypeptide or antigen-binding polypeptide complex described herein. Also provided herein is an immune cell comprising a CAR described herein.

[0041] Provided herein is a pharmaceutical composition comprising (i) an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody or antigen-binding fragment thereof, polypeptide, polynucleotide, vector, host cell, CAR, or immune cell described herein, or a combination thereof, and (ii) a pharmaceutically acceptable carrier.

[0042] Provided herein are kits that include an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody or antigen-binding fragment thereof, polypeptide, polynucleotide, vector, host cell, CAR, or immune cell described herein, or a combination thereof.

[0043] Also provided herein are certain methods of using the antigen-binding polypeptides, antigen-binding polypeptide complexes, antibodies or antigen-binding fragments thereof, polypeptides, polynucleotides, vectors, host cells, CARs, or immune cells, or combinations thereof, described herein. [Brief explanation of the drawings]

[0044] Some aspects of the present invention are herein described, by way of example only, with reference to the accompanying drawings. Referring now specifically to the drawings in detail, it is emphasized that the details shown are by way of example and for purposes of illustrative discussion of aspects of the invention.

[0045] [Figure 1A]The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 1B] The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 1C]The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 1D] The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 1E]The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 1F] The configurations of exemplary bispecific molecules of the invention are shown from the N-terminus to the C-terminus of the single-chain antigen-binding polypeptide(s). Figures 1A and 1D are bispecific molecules without an Fc region. Figure 1B is a bispecific tetravalent molecule with an Fc region. Figures 1C, 1E, and 1F are bispecific molecules with an Fc region. As used in Figures 1A-1F, VL1 refers to the first immunoglobulin light chain variable region, VL2 refers to the second immunoglobulin light chain variable region, VH1 refers to the first immunoglobulin heavy chain variable region, and VH2 refers to the second immunoglobulin heavy chain variable region. In Figures 1B, 1C, and 1F, CH2 refers to immunoglobulin heavy chain constant region 2, and CH3 refers to immunoglobulin heavy chain constant region 3. In Figures 1A and 1F, l1, l2, and l3 refer to amino acid linkers. In Figure 1D, L1, L2, and L3 refer to amino acid linkers. In Figures 1C and 1F, the circle symbols refer to knob-into-hole modifications. [Figure 2] FIG. 1B shows the SDS-PAGE results of the histidine-tagged nickel-NTA (Ni-NTA) purified bispecific molecule shown in FIG. 1A. [Figure 3]3A and 3B show the ELISA results for the bispecific molecule aCD19aCD38-His, which binds to CD19 (A) and CD38 (B), or an isotype control (control IgG). [Figure 4] FIG. 1B shows the SDS-PAGE results of Protein A purified bispecific tetravalent molecules with LALAPA Fc. [Figure 5] Figures 5A and 5B show ELISA results for bispecific tetravalent aCD28aCD3LALAPAFc, aCD3aCD28LALAPAFc, or an isotype control (control IgG) that binds to CD3 (A) and CD28 (B). The molecular structure is shown in Figure 1C. [Figure 6] 1 shows nuclear factor of activated T cells (NFAT) pathway activation by bispecific tetravalent aCD28aCD3L1LALAPAFc or aCD3aCD28L1LALAPAFc, or anti-CD3 and anti-CD28 mAbs using NFAT promoter-luciferase expressing human Jurkat T cells. [Figure 7] Figures 7A and 7B show ELISA results for bispecific aCD28aCD3LALAPAFc or aCD3aCD28LALAPAFc binding to CD3 (A) and CD28 (B), or an isotype control (control IgG). The molecular structure is shown in Figure 1C. [Figure 8] 8A-8C show exemplary tetraspecific molecule configurations of the invention. VL1 refers to a first immunoglobulin light chain variable region. VL2 refers to a second immunoglobulin light chain variable region. VL3 refers to a third immunoglobulin light chain variable region. VL4 refers to a fourth immunoglobulin light chain variable region. VH1 refers to a first immunoglobulin heavy chain variable region. VH2 refers to a second immunoglobulin heavy chain variable region. VH3 refers to a third immunoglobulin heavy chain variable region. VH4 refers to a fourth immunoglobulin heavy chain variable region. CH1 refers to immunoglobulin heavy chain constant region 1. CH2 refers to immunoglobulin heavy chain constant region 2. CH3 refers to immunoglobulin heavy chain constant region 3. CL refers to an immunoglobulin light chain constant region. The circle symbols in A-C refer to knob-into-hole modifications. [Figure 9] Figures 9A-9D show ELISA results for tetraspecific aCD28aCD3CD19CD38LALAPAFc, aCD3aCD28CD19CD38LALAPAFc, aCD28aCD3CD19CD38LALAPAFc, or aCD28aCD3CD38CD19LALAPAFc, which bind to CD3 (A), CD28 (B), CD19 (C), and CD38 (D), or an isotype control (control IgG). The molecular structure is shown in Figure 8A. [Figure 10] NFκB pathway activation by tetraspecific aCD28aCD3 / aCD19CD38L1LALAPAFc or aCD3aCD28 / CD19CD38L1LALAPAFc, or anti-CD3 mAb, using NFκB promoter-luciferase expressing human Jurkat T cells. [Figure 11] Figures 11A and 11B show activation of CD4+ (A) or CD8+ (B) T cells (CD69+) from three different donors with the tetraspecific molecules aCD28aCD3 / aCD19CD38L1LALAPAFc or aCD3aCD28 / CD19CD38L1LALAPAFc, or anti-CD3 mAb. [Figure 12] This shows that both the orientation and the linker can affect the expression of the tetraspecific molecule. [Figure 13] Figures 13A-13D show ELISA results for tetraspecific aCD28aCD3CD19CD38LALAPAFc with different linker lengths as shown in Figure 12, or an isotype control (control IgG), binding to CD3 (A), CD28 (B), CD19 (C), and CD38 (D). [Figure 14-1] Figures 14A-14D show ELISA results for tetraspecific aCD28aCD3CH1 / CD19CD38CL LALAPAFc with different linker lengths as shown in Figure 8B, which binds to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG). [Figure 14-2]Figures 14A-14D show ELISA results for tetraspecific aCD28aCD3CH1 / CD19CD38CL LALAPAFc with different linker lengths as shown in Figure 8B, which binds to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG). [Figure 15A] ELISA results for tetraspecific aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, or aCD3aCD28CD19CD38LALAPAFc, which bind to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG) are shown. The molecular structures are shown in Figure 8C. [Figure 15B] ELISA results for tetraspecific aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, or aCD3aCD28CD19CD38LALAPAFc, which bind to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG) are shown. The molecular structures are shown in Figure 8C. [Figure 15C] ELISA results for tetraspecific aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, or aCD3aCD28CD19CD38LALAPAFc, which bind to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG) are shown. The molecular structures are shown in Figure 8C. [Figure 15D]ELISA results for tetraspecific aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, aCD28aCD3CD38CD19LALAPAFc, or aCD3aCD28CD19CD38LALAPAFc, which bind to CD3 (A), CD28 (B), CD38 (C), and CD19 (D), or an isotype control (control IgG) are shown. The molecular structures are shown in Figure 8C. [Figure 15E] ELISA results for tetraspecific aCD28aCD3L1 / aCD38aCD19L1_HHLL, aCD28aCD3L1 / aCD19aCD38L1_HHLL, aCD3aCD28L1 / aCD38aCD19L1_HHLL, aCD3aCD28L1 / aCD19aCD38L1_HHLL, or an isotype control (control HuIgG) are shown, binding to CD3 (E), CD28 (F), CD38 (G), and CD19 (H). [Figure 15F] ELISA results for tetraspecific aCD28aCD3L1 / aCD38aCD19L1_HHLL, aCD28aCD3L1 / aCD19aCD38L1_HHLL, aCD3aCD28L1 / aCD38aCD19L1_HHLL, aCD3aCD28L1 / aCD19aCD38L1_HHLL, or an isotype control (control HuIgG) are shown, binding to CD3 (E), CD28 (F), CD38 (G), and CD19 (H). [Figure 15G] ELISA results for tetraspecific aCD28aCD3L1 / aCD38aCD19L1_HHLL, aCD28aCD3L1 / aCD19aCD38L1_HHLL, aCD3aCD28L1 / aCD38aCD19L1_HHLL, aCD3aCD28L1 / aCD19aCD38L1_HHLL, or an isotype control (control HuIgG) are shown, binding to CD3 (E), CD28 (F), CD38 (G), and CD19 (H). [Figure 15H]ELISA results for tetraspecific aCD28aCD3L1 / aCD38aCD19L1_HHLL, aCD28aCD3L1 / aCD19aCD38L1_HHLL, aCD3aCD28L1 / aCD38aCD19L1_HHLL, aCD3aCD28L1 / aCD19aCD38L1_HHLL, or an isotype control (control HuIgG) are shown, binding to CD3 (E), CD28 (F), CD38 (G), and CD19 (H). [Figure 16] 16A-16D show the configuration of exemplary bispecific molecules of the invention. VL1 refers to a first immunoglobulin light chain variable region. VL2 refers to a second immunoglobulin light chain variable region. VL3 refers to a third immunoglobulin light chain variable region. VL4 refers to a fourth immunoglobulin light chain variable region. VH1 refers to a first immunoglobulin heavy chain variable region. VH2 refers to a second immunoglobulin heavy chain variable region. VH3 refers to a third immunoglobulin heavy chain variable region. VH4 refers to a fourth immunoglobulin heavy chain variable region. CH3 refers to immunoglobulin heavy chain constant region 3. [Figure 17] Figures 17A-17E show exemplary configurations of trispecific antibody molecules of the invention. A: Bispecific arm paired with scFv-Fc. B: Bispecific arm paired with Fab-Fc. C: Bispecific arm paired with single-chain Fab (scFab). D: Bispecific arm paired with scFv single-chain CL-CH1-Fc. E: Bispecific arm fused to CH1 and paired with scFv-CL-Fc. [Figure 18A] ELISA results for the trispecific aCD28aCD3 / aCD38scFv, aCD28aCD3 / aCD38Fab, aCD28aCD3 / aCD38scFab, aCD28aCD3 / aCD38CLCH1, or an isotype control (control IgG) that binds to CD3 (Figure 18A), CD28 (Figure 18B), and CD38 (Figure 18C) are shown. Molecular structures are shown in Figures 17A-17D. [Figure 18B]ELISA results for the trispecific aCD28aCD3 / aCD38scFv, aCD28aCD3 / aCD38Fab, aCD28aCD3 / aCD38scFab, aCD28aCD3 / aCD38CLCH1, or an isotype control (control IgG) that binds to CD3 (Figure 18A), CD28 (Figure 18B), and CD38 (Figure 18C) are shown. Molecular structures are shown in Figures 17A-17D. [Figure 18C] ELISA results for the trispecific aCD28aCD3 / aCD38scFv, aCD28aCD3 / aCD38Fab, aCD28aCD3 / aCD38scFab, aCD28aCD3 / aCD38CLCH1, or an isotype control (control IgG) that binds to CD3 (Figure 18A), CD28 (Figure 18B), and CD38 (Figure 18C) are shown. Molecular structures are shown in Figures 17A-17D. [Figure 19] Activation (CD69+) of CD2+ T cells from three different donors with trispecific antibodies aCD28aCD3L1 / aCD38scFv, aCD3aCD28 / aCD38scFv, aCD28aCD3 / aCD38scFab, aCD3aCD28 / aCD38scFab, PMA / IO positive or negative isotype (control IgG) control is shown. [Figure 20] Figures 20A-20C show in vitro cytolysis of lymphoma tumor cells Z-138 by T cells mediated by the trispecific antibodies aCD28aCD3L1 / aCD38scFv, aCD3aCD28 / aCD38scFv, aCD28aCD3 / aCD38scFab, aCD3aCD28 / aCD38scFab, PMA / IO, or an isotype (control IgG) control from three different donors (A-C, respectively). [Figure 21A] ELISA results for the trispecific aCD28aCD3CL1CH1 / aCD38scFvCL, aCD28aCD3CL1CH1 / aCD19scFvCL, or an isotype control (control IgG) that binds to CD3 (Figure 21A), CD28 (Figure 21B), CD19 (Figure 21C), and CD38 (Figure 21D) are shown. The molecular structures are shown in Figure 17E. [Figure 21B]ELISA results for the trispecific aCD28aCD3CL1CH1 / aCD38scFvCL, aCD28aCD3CL1CH1 / aCD19scFvCL, or an isotype control (control IgG) that binds to CD3 (Figure 21A), CD28 (Figure 21B), CD19 (Figure 21C), and CD38 (Figure 21D) are shown. The molecular structures are shown in Figure 17E. [Figure 21C] ELISA results for the trispecific aCD28aCD3CL1CH1 / aCD38scFvCL, aCD28aCD3CL1CH1 / aCD19scFvCL, or an isotype control (control IgG) that binds to CD3 (Figure 21A), CD28 (Figure 21B), CD19 (Figure 21C), and CD38 (Figure 21D) are shown. The molecular structures are shown in Figure 17E. [Figure 21D] ELISA results for the trispecific aCD28aCD3CL1CH1 / aCD38scFvCL, aCD28aCD3CL1CH1 / aCD19scFvCL, or an isotype control (control IgG) that binds to CD3 (Figure 21A), CD28 (Figure 21B), CD19 (Figure 21C), and CD38 (Figure 21D) are shown. The molecular structures are shown in Figure 17E. [Figure 22] Non-limiting examples of different configurations of pentaspecific antibody molecules are shown below. vL1 is the first immunoglobulin light chain variable region. vL2 is the second immunoglobulin light chain variable region. vL3 is the third immunoglobulin light chain variable region. vL4 is the fourth immunoglobulin light chain variable region. vL5 is the fifth immunoglobulin light chain variable region. vH1 is the first immunoglobulin heavy chain variable region. vH2 is the second immunoglobulin heavy chain variable region. vH3 is the third immunoglobulin heavy chain variable region. vH4 is the fourth immunoglobulin heavy chain variable region. vH5 is the fifth immunoglobulin heavy chain variable region. CH2 is the immunoglobulin heavy chain constant region 2. CH3 is the immunoglobulin heavy chain constant region 3. The circle symbol in the CH3 region indicates a knob-into-hole modification. [Figure 23A]ELISA results for the pentaspecific aCD28aCD3LHaCD38 / aCD19aCD20, aCD28aCD3LHaCD38 / aCD20aCD19, aCD28aCD3HLaCD38 / aCD19aCD20, aCD28aCD3HLaCD38 / aCD20aCD19, or an isotype control (control IgG) that binds to CD3 (Figure 23A), CD28 (Figure 23B), CD38 (Figure 23C), and CD19 (Figure 23D). Molecular structures are shown in Figure 22. [Figure 23B] ELISA results for the pentaspecific aCD28aCD3LHaCD38 / aCD19aCD20, aCD28aCD3LHaCD38 / aCD20aCD19, aCD28aCD3HLaCD38 / aCD19aCD20, aCD28aCD3HLaCD38 / aCD20aCD19, or an isotype control (control IgG) that binds to CD3 (Figure 23A), CD28 (Figure 23B), CD38 (Figure 23C), and CD19 (Figure 23D). Molecular structures are shown in Figure 22. [Figure 23C] ELISA results for the pentaspecific aCD28aCD3LHaCD38 / aCD19aCD20, aCD28aCD3LHaCD38 / aCD20aCD19, aCD28aCD3HLaCD38 / aCD19aCD20, aCD28aCD3HLaCD38 / aCD20aCD19, or an isotype control (control IgG) that binds to CD3 (Figure 23A), CD28 (Figure 23B), CD38 (Figure 23C), and CD19 (Figure 23D). Molecular structures are shown in Figure 22. [Figure 23D] ELISA results for the pentaspecific aCD28aCD3LHaCD38 / aCD19aCD20, aCD28aCD3LHaCD38 / aCD20aCD19, aCD28aCD3HLaCD38 / aCD19aCD20, aCD28aCD3HLaCD38 / aCD20aCD19, or an isotype control (control IgG) that binds to CD3 (Figure 23A), CD28 (Figure 23B), CD38 (Figure 23C), and CD19 (Figure 23D). Molecular structures are shown in Figure 22. [Figure 24] 10 shows additional non-limiting examples of different configurations of tetraspecific antibody molecules. [Figure 25]1 shows an exemplary configuration of a masked tetraspecific antibody. The antibody variable domains (Fv) are shown as heavy / light chain pairs, with Fv1 through Fv3 targeting tumor-associated antigens (TAAs) or immune co-stimulatory receptors, and a fourth Fv targeting CD3 (αCD3 or αCD3). In some embodiments, the linker between Fv3 and αCD3 contains one or more protease recognition sites. [Figure 26] 1 shows SDS-PAGE results of in vitro cleavage of the exemplary masked tetraspecific molecules shown. The molecules were treated with either MTP or MMP9 protease as indicated. [Figure 27] ELISA binding results for exemplary masked tetraspecific molecules shown in Figure 26 with or without protease treatment, or a negative isotype (control IgG1) are shown. Molecules cleaved or not by the indicated MTP or MMP9 were tested for binding affinity to Trop2 and cMet. [Figure 28] ELISA binding results for exemplary masked tetraspecific molecules shown in Figure 26 with or without protease treatment, or a negative isotype (control IgG1) are shown. The molecules, cleaved or uncleaved by the indicated MTP or MMP9, were tested for binding affinity to CD28. [Figure 29] ELISA binding results for exemplary masked tetraspecific molecules shown in Figure 26 with or without protease treatment, or a negative isotype (control IgG1) are shown. Molecules cleaved or not by the indicated MTP or MMP9 were tested for binding affinity to CD3. [Figure 30] Figure 1 shows cytolysis of HCC1954 tumor cells by PBMCs (E:T:10:1) mediated by the exemplary masked tetraspecific molecule shown in Figure 2, or a negative isotype (control IgG1) from PBMCs of two donors (KP63250 and KP63251). [Figure 31]ELISA binding results of the indicated exemplary unmasked tetraspecific molecules, or a negative isotype (hIgG1LALPA) control, against their respective targets: hTrop2, hcMet, hCD28, and hCD3. [Figure 32] 1 shows CD69+ activation of CD2+ T cells from PBMCs of two different donors by an exemplary unmasked tetraspecific molecule or a negative isotype (IgG1LALPA) control. [Figure 33] 10 shows additional non-limiting examples of tetraspecific antibody molecules. [Figure 34A] Figure 34A shows a further non-limiting example of a tetravalent bispecific antibody construct called MX846. MX846 was analyzed for binding to CD3 by biolayer interferometry (BLI) (Figure 34B) and CD20 by flow cytometry (Figure 34C). [Figure 34B] Figure 34A shows a further non-limiting example of a tetravalent bispecific antibody construct called MX846. MX846 was analyzed for binding to CD3 by biolayer interferometry (BLI) (Figure 34B) and CD20 by flow cytometry (Figure 34C). [Figure 34C] Figure 34A shows a further non-limiting example of a tetravalent bispecific antibody construct called MX846. MX846 was analyzed for binding to CD3 by biolayer interferometry (BLI) (Figure 34B) and CD20 by flow cytometry (Figure 34C). [Figure 35A] Figure 35A shows a further non-limiting example of a tetravalent trispecific antibody construct called MX855. MX855 was analyzed for binding to CD3 and CD28 by biolayer interferometry (BLI) (Figure 35B), and for binding to CD20 by flow cytometry (Figure 35C). [Figure 35B]Figure 35A shows a further non-limiting example of a tetravalent trispecific antibody construct called MX855. MX855 was analyzed for binding to CD3 and CD28 by biolayer interferometry (BLI) (Figure 35B), and for binding to CD20 by flow cytometry (Figure 35C). [Figure 35C] Figure 35A shows a further non-limiting example of a tetravalent trispecific antibody construct called MX855. MX855 was analyzed for binding to CD3 and CD28 by biolayer interferometry (BLI) (Figure 35B), and for binding to CD20 by flow cytometry (Figure 35C). [Figure 36A] Figure 36A shows a further non-limiting example of a tetraspecific antibody construct called MX851. MX851 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 36B), and for binding to CD20 by flow cytometry (Figure 36C). [Figure 36B] Figure 36A shows a further non-limiting example of a tetraspecific antibody construct called MX851. MX851 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 36B), and for binding to CD20 by flow cytometry (Figure 36C). [Figure 36C] Figure 36A shows a further non-limiting example of a tetraspecific antibody construct called MX851. MX851 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 36B), and for binding to CD20 by flow cytometry (Figure 36C). [Figure 37A] Figure 37A shows a further non-limiting example of a tetraspecific antibody construct called MX853. MX853 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 37B), and CD20 by flow cytometry (Figure 37C). [Figure 37B]Figure 37A shows a further non-limiting example of a tetraspecific antibody construct called MX853. MX853 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 37B), and CD20 by flow cytometry (Figure 37C). [Figure 37C] Figure 37A shows a further non-limiting example of a tetraspecific antibody construct called MX853. MX853 was analyzed for binding to CD3, CD28, and BCMA by biolayer interferometry (BLI) (Figure 37B), and CD20 by flow cytometry (Figure 37C). [Figure 38] Figures 38A and 38B show T cell killing of mantle cell lymphoma cell line Z-138 mediated by tetravalent tetraspecific MX851 (A) and tetravalent trispecific MX855 (B). [Figure 39A] Figure 39A shows a further non-limiting example of a trispecific antibody construct called MX894 (VRC01scFv / PGT121x10e8v4L1IgG1LS). MX894 was analyzed by biolayer interferometry (BLI) for binding to the 10e8 fusion peptide (Figure 39B), and to CD4 site-dependent (Figure 39C) and CD4 site-independent (Figure 39D) HIV spike protein. [Figure 39B] Figure 39A shows a further non-limiting example of a trispecific antibody construct called MX894 (VRC01scFv / PGT121x10e8v4L1IgG1LS). MX894 was analyzed by biolayer interferometry (BLI) for binding to the 10e8 fusion peptide (Figure 39B), and to CD4 site-dependent (Figure 39C) and CD4 site-independent (Figure 39D) HIV spike protein. [Figure 39C]Figure 39A shows a further non-limiting example of a trispecific antibody construct called MX894 (VRC01scFv / PGT121x10e8v4L1IgG1LS). MX894 was analyzed by biolayer interferometry (BLI) for binding to the 10e8 fusion peptide (Figure 39B), and to CD4 site-dependent (Figure 39C) and CD4 site-independent (Figure 39D) HIV spike protein. [Figure 39D] Figure 39A shows a further non-limiting example of a trispecific antibody construct called MX894 (VRC01scFv / PGT121x10e8v4L1IgG1LS). MX894 was analyzed by biolayer interferometry (BLI) for binding to the 10e8 fusion peptide (Figure 39B), and to CD4 site-dependent (Figure 39C) and CD4 site-independent (Figure 39D) HIV spike protein. [Figure 40A] Figure 40A shows a further non-limiting example of a tetraspecific antibody construct designated MX873 (VRC26.25x10-1074L9 / VRC01xPGT121L1 IgG1LS). MX873 was analyzed for CD4 site-dependent (Figure 40B) and CD4 site-independent (Figure 40C) binding to the HIV spike protein by biolayer interferometry (BLI). [Figure 40B] Figure 40A shows a further non-limiting example of a tetraspecific antibody construct designated MX873 (VRC26.25x10-1074L9 / VRC01xPGT121L1 IgG1LS). MX873 was analyzed for CD4 site-dependent (Figure 40B) and CD4 site-independent (Figure 40C) binding to the HIV spike protein by biolayer interferometry (BLI). [Figure 40C] Figure 40A shows a further non-limiting example of a tetraspecific antibody construct designated MX873 (VRC26.25x10-1074L9 / VRC01xPGT121L1 IgG1LS). MX873 was analyzed for CD4 site-dependent (Figure 40B) and CD4 site-independent (Figure 40C) binding to the HIV spike protein by biolayer interferometry (BLI). [Figure 41A]Figure 41A shows a further non-limiting example of a tetraspecific antibody construct called MX875 (10-1074xVRC26.25L9 / VRC01xPGT121L1 IgG1LS). MX875 was analyzed by biolayer interferometry (BLI) for binding to the CD4 site-dependent (Figure 41B) and CD4 site-independent (Figure 41C) HIV spike protein. [Figure 41B] Figure 41A shows a further non-limiting example of a tetraspecific antibody construct called MX875 (10-1074xVRC26.25L9 / VRC01xPGT121L1 IgG1LS). MX875 was analyzed by biolayer interferometry (BLI) for binding to the CD4 site-dependent (Figure 41B) and CD4 site-independent (Figure 41C) HIV spike protein. [Figure 41C] Figure 41A shows a further non-limiting example of a tetraspecific antibody construct called MX875 (10-1074xVRC26.25L9 / VRC01xPGT121L1 IgG1LS). MX875 was analyzed by biolayer interferometry (BLI) for binding to the CD4 site-dependent (Figure 41B) and CD4 site-independent (Figure 41C) HIV spike protein. [Figure 42A] Figure 42A shows a further non-limiting example of a tetraspecific antibody construct called MX877 (STAR_VRC26.25xPGT128L9 / STAR_VRC01xPGT121L1 IgG1LS). MX877 was analyzed for CD4 site-dependent (Figure 42B) and CD4 site-independent (Figure 42C) binding to the HIV spike protein by biolayer interferometry (BLI). [Figure 42B] Figure 42A shows a further non-limiting example of a tetraspecific antibody construct called MX877 (STAR_VRC26.25xPGT128L9 / STAR_VRC01xPGT121L1 IgG1LS). MX877 was analyzed for CD4 site-dependent (Figure 42B) and CD4 site-independent (Figure 42C) binding to the HIV spike protein by biolayer interferometry (BLI). [Figure 42C]Figure 42A shows a further non-limiting example of a tetraspecific antibody construct called MX877 (STAR_VRC26.25xPGT128L9 / STAR_VRC01xPGT121L1 IgG1LS). MX877 was analyzed for CD4 site-dependent (Figure 42B) and CD4 site-independent (Figure 42C) binding to the HIV spike protein by biolayer interferometry (BLI). DETAILED DESCRIPTION OF THE INVENTION

[0046] Detailed Description of the Invention The present invention is directed to antigen-binding polypeptides and antigen-binding polypeptide complexes (e.g., antibodies or antigen-binding fragments thereof) with improved characteristics. In some aspects, the present invention allows for the generation of multispecific and multifunctional antigen-binding polypeptides and antigen-binding polypeptide complexes through the expression of complementary self-assembling heavy and light chains expressed with a single polypeptide per arm, and, optionally, the addition of specific amino acid linkers. Because of this multifunctionality, the antigen-binding polypeptides and antigen-binding polypeptide complexes of the present invention can bind to specific combinations of target molecules for selectivity or amplification / neutralization, bring together two or more cell types, bring together targets to deliver activation signals, modify the disease microenvironment, and improve binding affinity for improved efficacy.

[0047] Throughout the specification and claims, various terms relating to aspects of the present disclosure are used. Unless otherwise indicated, such terms shall be given their ordinary meaning in the art. Other terms that are specifically defined shall be construed in a manner consistent with the definition set forth herein.

[0048] definition As used herein, the term "antigen-binding polypeptide" refers to a polypeptide that has the ability to specifically bind to one or more substances (i.e., one or more antigens or epitopes) that induce an immune response.

[0049] As used herein, the term "antigen-binding polypeptide complex" refers to a group of two, three, four, or more associated polypeptides, where at least one polypeptide has the ability to specifically bind to one or more antigens. Antigen-binding polypeptide complexes include, but are not limited to, antibodies or antigen-binding fragments thereof.

[0050] The term "antibody" includes, but is not limited to, a glycoprotein immunoglobulin that specifically binds an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region comprises three constant domains, CH1, CH2, and CH3. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region comprises one constant domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity-determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FRs). Each VH and VL comprises three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of the antibody can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (C1q) of the classical complement system. The heavy chain may or may not have a C-terminal lysine. Unless otherwise specified herein, amino acids in the variable regions are numbered using the Kabat numbering system, and amino acids in the constant regions are numbered using the EU system.

[0051] The term "monoclonal antibody," as used herein, refers to an antibody produced by a single clone of B cells and that binds to the same epitope. In contrast, the term "polyclonal antibody" refers to a population of antibodies produced by different B cells and that bind to different epitopes of the same antigen. The term "antibody" includes, by way of example, monoclonal and polyclonal antibodies, chimeric and humanized antibodies, human or non-human antibodies, fully synthetic antibodies, and single-chain antibodies. Non-human antibodies can be humanized by recombinant methods to reduce their immunogenicity in humans.

[0052] The antibody may be an altered antibody (e.g., by mutation, deletion, substitution, conjugation to a non-antibody moiety). For example, the antibody may contain one or more variant amino acids that alter the properties (e.g., functional properties) of the antibody (compared to a naturally occurring antibody). For example, several such alterations are known in the art, affecting, for example, half-life, effector functions, and / or the immune response to the antibody in a patient. The term antibody also includes artificial polypeptide constructs that contain an antigen-binding site derived from at least one antibody.

[0053] An "antigen-binding fragment" of an antibody refers to one or more fragments or portions of an antibody that retain the ability to specifically bind to the antigen bound by the whole antibody. It has been shown that the antigen-binding function of an antibody can be performed by fragments or portions of a full-length antibody. An antigen-binding fragment may contain the antigen-determining region (e.g., complementarity-determining region (CDR)) of the intact antibody. Examples of antigen-binding fragments of antibodies include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, and single-chain antibodies. Antigen-binding fragments of antibodies can be derived from any animal species, such as rodents (e.g., mice, rats, or hamsters) and humans, or can be artificially produced.

[0054] Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked by a synthetic linker that allows them to be produced using recombinant methods as a single protein chain (known as a single-chain Fv (scFv), see, e.g., Bird et al. (1988) Science 242:423-426, and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883), allowing the VL and VH regions to pair to form a monovalent molecule. Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding fragment" of an antibody.

[0055] Antigen-binding fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. Antigen-binding fragments can be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact immunoglobulins.

[0056] As used herein, the term "variable region" typically refers to a portion of an antibody, generally a portion of either the light or heavy chain, typically the amino-terminal approximately 110-120 amino acids, or 110-125 amino acids in mature heavy chains and approximately 90-115 amino acids in mature light chains, which vary significantly in sequence among antibodies and are responsible for the binding and specificity of a particular antibody to its particular antigen. Sequence variability is concentrated in these regions, called complementarity-determining regions (CDRs), while more highly conserved regions within the variable domain are called framework regions (FRs). While not wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with the antigen. In some embodiments, the variable region is a mammalian variable region, e.g., a human, mouse, or rabbit variable region. In some embodiments, the variable region comprises rodent or mouse CDRs and human framework regions (FRs). In some embodiments, the variable region is a primate (e.g., non-human primate) variable region. In some embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0057] As used herein, the term "complementarity determining region" or "CDR" refers to each of the regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops (hypervariable loops) and / or contain antigen-contacting residues. An antibody may contain six CDRs, e.g., three in the VH and three in the VL.

[0058] The terms "VL," "VL region," and "VL domain" are used interchangeably herein to refer to the light chain variable region of an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof. In some aspects, a VL region is referred to herein as VL1, which denotes a first light chain variable region, VL2, which denotes a second light chain variable region, VL3, which denotes a third light chain variable region, VL4, which denotes a fourth light chain variable region, etc. A listed VL region (e.g., VL1) may have the same or different antigen-binding characteristics and / or the same or different sequence as another listed VL region (e.g., VL2).

[0059] The terms "VH," "VH region," and "VH domain" are used interchangeably herein to refer to the heavy chain variable region of an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof. In some aspects, a VH region is referred to herein as VH1, which denotes a first heavy chain variable region, VH2, which denotes a second heavy chain variable region, VH3, which denotes a third heavy chain variable region, VH4, which denotes a fourth heavy chain variable region, etc. A listed VH region (e.g., VH1) may have the same or different antigen-binding characteristics, and / or the same or different sequence, as another listed VH region (e.g., VH2).

[0060] As used herein, "Kabat numbering" and similar terms are art-recognized and refer to a numbering system for amino acid residues in the heavy and light chain variable regions of an antibody or antigen-binding fragment thereof. In some embodiments, CDRs can be determined according to the Kabat numbering system (see, e.g., Kabat EA & Wu TT (1971) Ann NY Acad Sci 190:382-391 and Kabat EA et al., (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). Using the Kabat numbering system, the CDRs in an antibody heavy chain molecule are typically located at amino acid positions 31-35, followed by 35 (designated 35A and 35B in the Kabat numbering scheme) (CDR1), which may optionally contain one or two additional amino acids, at amino acid positions 50-65 (CDR2), and at amino acid positions 95-102 (CDR3). Using the Kabat numbering system, the CDRs in an antibody light chain molecule are typically located at amino acid positions 24-34 (CDR1), at amino acid positions 50-56 (CDR2), and at amino acid positions 89-97 (CDR3).

[0061] As used herein, the terms "constant region" or "constant domain" are used interchangeably to refer to a portion of an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof, e.g., the carboxyl-terminal portion of the light chain and / or heavy chain, which is not directly involved in binding the antibody to an antigen but can exhibit various effector functions, such as interacting with the Fc region. Constant regions generally have a more conserved amino acid sequence compared to the variable region. In some embodiments, an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof comprises a constant region, or portion thereof, sufficient for antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC).

[0062] As used herein, the terms "fragment crystallizable region," "Fc region," or "Fc domain" are used interchangeably herein to refer to the tail region of an antibody that interacts with cell surface receptors called Fc receptors and several proteins of the complement system. The Fc region typically includes the CH2 and CH3 regions and, optionally, an immunoglobulin hinge. Examples of Fc regions include, but are not limited to, the amino acid sequence of any one of SEQ ID NOs: 391-404, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 391-404. Examples of CH2 regions include, but are not limited to, the amino acid sequence of any one of SEQ ID NOs: 410-415, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 410-415. Examples of CH3 regions include, but are not limited to, the amino acid sequence of any one of SEQ ID NOs: 416-419, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 416-419.

[0063] As used herein, the terms "immunoglobulin hinge," "hinge," "hinge domain," or "hinge region" are used interchangeably to refer to the region of an antigen-binding polypeptide, antigen-binding polypeptide complex, or heavy chain between the Fab and Fc portions of an antibody or antigen-binding fragment thereof. The hinge provides structure, positioning, and flexibility, aiding in the normal function of the antibody (e.g., for cross-linking two antigens or binding two antigenic determinants on the same antigen molecule). Immunoglobulin hinges are divided into upper, middle, and lower hinge regions, which can be separated based on structural and / or genetic components. Immunoglobulin hinges of the present invention may contain one, two, or all three of these regions. Structurally, the upper hinge region extends from the C-terminal end of CH1 to the first hinge disulfide bond. The middle hinge region extends from the first cysteine ​​to the last cysteine ​​in the hinge. The lower hinge region extends from the last cysteine ​​to the glycine of CH2. The cysteines present in the hinge form the interchain disulfide bonds that link the immunoglobulin monomers.

[0064] As used herein, the term "Fab" refers to the region of an antibody that binds to an antigen, typically consisting of one constant domain and one variable domain from each of the heavy and light chains.

[0065] As used herein, the term "heavy chain" refers to an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof, typically composed of a heavy chain variable region (VH), heavy chain constant region 1 (CH1), heavy chain constant region 2 (CH2), and heavy chain constant region 3 (CH3). A typical antibody is composed of two heavy chains and two light chains. When used in reference to an antibody, heavy chain can refer to any distinct type, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the constant region, which gives rise to antibodies of the IgA, IgD, IgE, IgG, and IgM classes, respectively, including subclasses of IgG, e.g., IgG1, IgG2, IgG3, and IgG4. Heavy chain amino acid sequences are known in the art. In some embodiments, the heavy chain is a human heavy chain.

[0066] As used herein, the term "light chain" refers to a portion of an antigen-binding polypeptide, antigen-binding polypeptide complex, antibody, or antigen-binding fragment thereof, typically composed of a light chain variable region (VL) and a light chain constant region (CL). A typical antibody is composed of two light chains and two heavy chains. When used in reference to an antibody, light chain may refer to any distinct type, e.g., kappa (κ) or lambda (λ), based on the amino acid sequence of the constant region. Light chain amino acid sequences are known in the art. In some embodiments, the light chain is a human light chain.

[0067] The term "chimeric" antibody or antigen-binding fragment thereof refers to an antibody or antigen-binding fragment thereof whose amino acid sequences are derived from two or more species. Typically, the variable regions of both the light and heavy chains correspond to the variable regions of an antibody or antigen-binding fragment thereof from one species of mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and capacity, while the constant regions are homologous to the sequences of an antibody or antigen-binding fragment thereof from another species (usually human) to avoid eliciting an immune response in that species.

[0068] The term "humanized" antibody or antigen-binding fragment thereof refers to a form of a non-human (e.g., murine) antibody or specific immunoglobulin chain antigen-binding fragment, chimeric immunoglobulin, or fragment thereof that contains minimal non-human (e.g., murine) sequence. Typically, humanized antibodies or antigen-binding fragments thereof are human immunoglobulins in which residues from the complementarity-determining regions (CDRs) are replaced by residues from the CDRs of a non-human species (e.g., mouse, rat, rabbit, hamster) having the desired specificity, affinity, and capacity (Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988)). In some embodiments, Fv framework region (FR) residues of a human immunoglobulin are replaced with corresponding residues in an antibody or fragment from a non-human species having the desired specificity, affinity, and capacity. To improve and optimize the specificity, affinity, and / or capacity of the antibody or antigen-binding fragment, the humanized antibody or antigen-binding fragment can be further modified by substitution of additional residues within the Fv framework regions and / or the replaced non-human residues. Generally, a humanized antibody or antigen-binding fragment will comprise substantially all of at least one, typically two or three, variable domains containing all or substantially all of the CDR regions corresponding to the non-human immunoglobulin, while all or substantially all of the FR regions are of human immunoglobulin consensus sequences. The humanized antibody or antigen-binding fragment may also comprise at least a portion of a constant region, typically that of a human immunoglobulin. Examples of methods used to generate humanized antibodies are known and are described, for example, in U.S. Pat. No. 5,225,539, Roguska et al., Proc. Natl. Acad. Sci., USA, 91(3):969-973 (1994), and Roguska et al., Protein Eng. 9(10):895-904 (1996).

[0069] As used herein, the term "human" antibody or antigen-binding fragment thereof means an antibody or antigen-binding fragment thereof having an amino acid sequence derived from the human immunoglobulin locus, and such an antibody or antigen-binding fragment is produced using recombinant techniques known in the art. This definition of a human antibody or antigen-binding fragment thereof includes intact or full-length antibodies and fragments thereof.

[0070] An "isolated" polypeptide, polypeptide complex, antibody, antigen-binding fragment thereof, polynucleotide, vector, or host cell is a polypeptide, polypeptide complex, antibody, antigen-binding fragment thereof, polynucleotide, vector, or host cell that is in a form not found in nature. Isolated polypeptides, polypeptide complexes, antibodies, antigen-binding fragments thereof, polynucleotides, vectors, or host cells include those that have been purified to the extent that they are no longer in the form in which they are found in nature. In some embodiments, an isolated polypeptide, polypeptide complex, antibody, antigen-binding fragment thereof, polynucleotide, vector, or host cell is substantially pure. As used herein, "substantially pure" refers to material that is at least 50% pure (i.e., free from contaminants), at least 90% pure, at least 95% pure, at least 99% pure, or at least 98% pure.

[0071] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. Polymers can be linear or branched, can comprise modified amino acids, and can be interrupted by non-amino acids. These terms also encompass amino acid polymers, whether naturally occurring or modified by intervention, e.g., disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are polypeptides containing, for example, one or more analogs of an amino acid (including, e.g., unnatural amino acids), as well as other modifications known in the art. Because the polypeptides of the present invention are based on antibodies, it is understood that in some aspects the polypeptides can occur as single chains or associated chains.

[0072] The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the indefinite article "a" or "an" should be understood to refer to "one or more" of any shown or listed components.

[0073] As used herein, the term "and / or" is to be construed as a specific disclosure of each of the two specified features or components, with or without the other features or components. Thus, as used herein, the term "and / or" when used in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, the term "and / or" when used in phrases such as "A, B, and / or C" is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0074] In all cases, embodiments are described herein using language such as "comprising," "having," and the like, but analogous embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0075] As used herein, the term "about" refers to a value or composition that is within an acceptable error range for a particular value or composition, as determined by one of ordinary skill in the art, and will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within 1 or more standard deviations, as is customary in the art. Alternatively, "about" can mean a range of up to 10% or 20% (i.e., ±10% or ±20%). For example, approximately 3 mg can refer to any number between 2.7 mg and 3.3 mg (10%), or between 2.4 mg and 3.6 mg (20%). Furthermore, specifically with respect to biological systems or processes, the term can mean up to an order of magnitude, or up to five times the value. When a particular value or composition is provided in this application and claims, unless otherwise specified, the meaning of "about" should be assumed to be within an acceptable error range for that particular value or composition.

[0076] As described herein, any numerical range, concentration range, percentage range, ratio range, or integer range, unless otherwise indicated, is understood to include any integer value within the stated range, and, where appropriate, fractions thereof (such as integer tenths and hundredths).

[0077] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press, The Dictionary of Cell and Molecular Biology, 5th ed., 2013, Academic Press, and the Oxford Dictionary of Biochemistry and Molecular Biology, 2006, Oxford University Press provide those skilled in the art with a general dictionary of many of the terms used in this disclosure.

[0078] Units, prefixes, and symbols are denoted in the format accepted by the Systeme International de Unites (SI). Numerical ranges are intended to be inclusive of the numbers defining the range. The headings provided herein are not limitations of the various aspects of the disclosure, which may be had by reference to the specification as a whole. Accordingly, terms defined herein are more fully defined by reference to the specification as a whole.

[0079] Various aspects are described in further detail in the following paragraphs.

[0080] Antigen-binding polypeptides and antigen-binding polypeptide complexes In some aspects, the present invention is directed to antigen-binding polypeptides and antigen-binding polypeptide complexes that have certain structural characteristics.

[0081] In some embodiments, the present invention is directed to antigen-binding polypeptides and antigen-binding polypeptide complexes having a structure represented by VL1-VL2-VH2-VH1 or VH1-VH2-VL2-VL1. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex contains an amino acid linker between any two regions depicted in the structures described herein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex may contain an Fc region, a CH1 region, a CL region, a CH3 region, or any combination thereof. In some embodiments, the Fc region, CH1 region, CL region, and / or CH3 region are located at the carboxy terminus of the antigen-binding polypeptide and are optionally linked to the polypeptide by at least one amino acid linker. In some embodiments, the Fc region comprises the amino acid sequence of any one of SEQ ID NOs: 391-404, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 391-404. In some embodiments, the CH1 region comprises the amino acid sequence of any one of SEQ ID NOs: 405-409, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 405-409. In some embodiments, the CL region comprises the amino acid sequence of SEQ ID NO: 420 or 421, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 420 or 421. In some embodiments, the antigen-binding polypeptide complex is an antibody or antigen-binding fragment thereof.

[0082] In some aspects, the antigen-binding polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, and L1, L2, and L3 are amino acid linkers. In some aspects, the antigen-binding polypeptide has a structure represented by VL1-VL2-VH2-VH1. In some aspects, the antigen-binding polypeptide has a structure represented by VH1-VH2-VL2-VL1. In some embodiments, the antigen-binding polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1. In some embodiments, the antigen-binding polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1.

[0083] In some aspects, the antigen-binding polypeptide complex comprises a first polypeptide and a second polypeptide, wherein the first polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2- VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, and L1, L2, and L3 are amino acid linkers. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, or VH1-L1-VH2-L2-VL2-L3-VL1.

[0084] In some embodiments, the antigen-binding polypeptide further comprises at least one Fc region, optionally located at its carboxy terminus. The Fc region may be linked to the polypeptide via at least one amino acid linker. For example, the antigen-binding polypeptide may have a structure represented by VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, and L4 are amino acid linkers.

[0085] In some aspects, the antigen-binding polypeptide complexes defined herein further comprise at least one Fc region, optionally located at the carboxy terminus of the first polypeptide and / or the second polypeptide. The Fc region may be linked to the first polypeptide and / or the second polypeptide via at least one amino acid linker. For example, the antigen-binding polypeptide complexes may comprise a first polypeptide and a second polypeptide, wherein the first polypeptide has a sequence similar to: VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1- L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH 1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, and L4 are amino acid linkers.In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc.

[0086] In some embodiments, the antigen-binding polypeptide further comprises at least one CH1 region and / or CL region, optionally located at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CH1-CL at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CL-CH1 at its carboxy terminus. The CH1 region and / or CL region may be linked to the polypeptide via at least one amino acid linker. When both a CH1 region and a CL region are present, they may be linked to each other via at least one amino acid linker. For example, the antigen-binding polypeptide can be any of the following: VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-CL, VH ... 1-VH2-L2-VL2-L3-VL1-L4-CH1-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-CH1, VH1-L1-VH 2-L2-VL2-L3-VL1-L4-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2 -L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0087] In some aspects, the antigen-binding polypeptide complex defined herein further comprises at least one CH1 region and / or CL region, optionally located at the carboxy terminus of the first polypeptide and / or second polypeptide. For example, the carboxy terminus of the first polypeptide and / or second polypeptide may comprise a CH1 region. For example, the carboxy terminus of the first polypeptide and / or second polypeptide may comprise a CL region. For example, the carboxy terminus of the first polypeptide and / or second polypeptide may comprise both a CH1 and a CL region. In some aspects, the carboxy terminus of the first polypeptide and / or second polypeptide comprises the structure CH1-CL. In some aspects, the carboxy terminus of the first polypeptide and / or second polypeptide comprises the structure CL-CH1. The CH1 region and / or CL region may be linked to the first polypeptide and / or second polypeptide via at least one amino acid linker. When both a CH1 region and a CL region are present, they may be linked to each other via at least one amino acid linker. For example, an antigen-binding complex can comprise a first polypeptide and a second polypeptide, wherein the first polypeptide is VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2 -L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL , VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1,or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, and the second polypeptide is VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1- CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH 2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH By 1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1 wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is a light chain constant region, and L1, L2, L3, L4, and L5 are amino acid linkers. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1 and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1,VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL 2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3- and VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1 and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1 -L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1 In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VL2-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL,VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-V L1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4- CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1 -L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1 In some embodiments, the first polypeptide has a structure represented by: VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-C H1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-C L, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L It has a structure represented by 4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL -CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL,In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-CH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VL2-VL1-CH1, VL1-VL2-VH, 2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH 2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CL-CH1 and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4- and having a structure represented by CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4- and having a structure represented by CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4- and having a structure represented by CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4- and having a structure represented by CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2- L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2- L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2- L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL. 、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3- VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2 -L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、V It has the structure shown by L1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1.

[0088] In some embodiments, the antigen-binding polypeptide further comprises at least two of an Fc region, a CH1 region, and a CL region, optionally located at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CH1-Fc at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CL-Fc at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CL-CH1-Fc at its carboxy terminus. In some embodiments, the antigen-binding polypeptide further comprises the structure CH1-CL-Fc at its carboxy terminus. The Fc region, CH1 region, and / or CL region may be linked to the polypeptide via at least one amino acid linker. The Fc region, CH1 region, and / or CL region may be linked to each other via at least one amino acid linker.For example, the C-terminal structure is VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-C L-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL 1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2- VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH VL1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, where VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, and VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. 1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0089] In some aspects, the antigen-binding polypeptide complex defined herein further comprises at least two of an Fc region, a CH1 region, and a CL region, optionally located at the carboxy terminus of the first polypeptide and / or the second polypeptide. For example, the carboxy terminus of the first polypeptide and / or the second polypeptide may comprise the structure CH1-Fc. For example, the carboxy terminus of the first polypeptide and / or the second polypeptide may comprise the structure CL-Fc. For example, the carboxy terminus of the first polypeptide and / or the second polypeptide may comprise the structure CL-CH1-Fc. For example, the carboxy terminus of the first polypeptide and / or the second polypeptide may comprise the structure CH1-CL-Fc. In some aspects, the first polypeptide may comprise at least two of an Fc region, a CH1 region, and a CL region at its C-terminus, and the second polypeptide may comprise an Fc region at its C-terminus. The Fc region, CH1 region, and / or CL region can be linked to the first polypeptide and / or the second polypeptide via at least one amino acid linker. The Fc region, CH1 region, and / or CL region can be linked to each other via at least one amino acid linker. For example, an antigen-binding complex can include a first polypeptide and a second polypeptide, wherein the first polypeptide has a sequence similar to: VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-C H1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc,and the second polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, and the second polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-VH2-L2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH ... -VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH 1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1 -VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, wherein VL1 is a first immunoglobulin light chain VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers. In some aspects, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-Fc, and the second polypeptide has a structure represented by Fc,VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2- VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3 -VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、V H1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-FcにIn some embodiments, polypeptide 1 has a structure represented by VH1-VH2-VL2-VL1-CH1-Fc, polypeptide 2 has Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1- CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1 -Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-Fc, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL ... -VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL 2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL -Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5- CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CL-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2 -VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2- VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc,VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH 1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-F In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-CL-Fc, and the second polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1- VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1 -VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5 In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-CL-Fc and the second polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2- L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-V L2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1- VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L It has a structure represented by 5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-CH1-Fc, and the second polypeptide has a structure represented by any of Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1- VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1 -VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5 -CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CL-CH1-Fc, and the second polypeptide has a structure represented by any of Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1- VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1 -VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5 -CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-F c, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-F c, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3- VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1- L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-F c, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-F c, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3- VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1- L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, c, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-F c, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3- VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1- L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, c, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-F c, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3- VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1- L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1 -CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1- CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CH1-Fc, VL1-VL2-VH2-VH1 -CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1- CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc and the second polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, VL1-VL2-L2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc. 2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc. CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH 1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VL1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CH1-Fc, VL1-VL2-VH2-VH1 -CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1- CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2- L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.

[0090] In some embodiments, the antigen-binding polypeptide complex contains the 1st polypeptide and the 2nd polypeptide. L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1- VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2- VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L 3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-V H2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL 1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-C H1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L 3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL It has the structure shown by 2-L3-VL1-L4-CL-L5-CH1-Fc. VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-Fc L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL L1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、 VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-C H1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-V L2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1 -Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3- VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1- VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, and VH1 is is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH1 is immunoglobulin heavy chain constant region 1, CL is an immunoglobulin light chain constant region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1 and the second polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc,VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL 、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-C H1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL 2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2 -L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L 3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL 1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or having a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1, and the second polypeptide has Fc. VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL 1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL 2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1- VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH 2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、 VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、 VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-V It has a structure represented by L2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the 1st polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1, the 2nd polypeptide has a Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3 -VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VH1 L1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-V H2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L 3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-CL L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-C L-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-V H2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-V It has a structure represented by L1-L4-CL-L5-CH1-Fc. In some embodiments, the 1st polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1, the 2nd polypeptide has a structure represented by Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2 -L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1 H1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2- L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc 、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc c. 3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4- It has a structure represented by CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-Fc. 2のpolypeptideは、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2 -VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-C L、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2- L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5 -CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1- Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1 -L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL- It has the structure represented by Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.VH1-VH2-VL2-VL1-VH1-VL2-VL1-VL1-VL1-VL2-VL1-VL1-VL2-VL2-VL1-VL1-VL2-VL2-VL1 VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4 -Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2 -VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L 2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L3-VL1-L4-CL L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1 -CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc Fc.VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L It has a structure represented by 3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc,VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、 VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2- The structure represented by VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc In some embodiments, the 1st polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-Fc. L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2 -L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH 2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-V H2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1 -VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-V H1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1- VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1 -L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc has a structure. In some embodiments, the 1st polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc.プチドは、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1 1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-C L、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2- L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5 -CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1- Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1 -L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL- It has the structure represented by Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.It has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc,VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5- It has a structure represented by CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1.リpeptideは、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2 VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc c、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VL2-VL2-VL1-CH1-C L、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2- L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc c.It has a structure represented by VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL,VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-V H2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-C H1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4- CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-V Represented by H2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL, the second polypeptide has Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2 -L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1- Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2 -VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1 -L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-C L-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1- VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-C H1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4- CL-L5-CH1-Fc has a structure represented by CL-L5-CH1-Fc. In some embodiments, polypeptide 1 has a structure represented by VH1-VH2-VL2-VL1-CL, polypeptide 2 has a structure represented by Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1- VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1 VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、 VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1- CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-V L1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1 VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-C L-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1- VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2- VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fcrepresented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, In some embodiments, polypeptide 1 has a structure represented by VL1-VL2-VH2-VH1-CH1-CL, polypeptide 2 has Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2- L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3 -VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH 1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL 2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1- VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2- L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH 1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、orVH1-L1-VH2-L 2-VL2-L3-VL1-L4-CL-L5-CH1-CL has a structure represented by Fc. In some embodiments, the 1st polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-CL, and the 2nd polypeptide has a structure represented by Fc, VL1-VL 2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL1 VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-V H2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL 2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、It has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-CH1, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL,VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-V L1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2- VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、 VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VH2-VL2-VL1-CL-Fc L2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1 -VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3- It has several structures represented by VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In an embodiment, the 1st polypeptide has the structure represented by VH1-VH2-VL2-VL1-CL-CH1, the 2nd polypeptide has the structure Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1,VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, V L1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、V L1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-FcIn some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. - It has a structure represented by -L2-VH2-L3-VH1-L4-CH1, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc,Structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VH1-L4-CL-L5-CH1-Fc In some embodiments, polypeptide 1 has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, polypeptide 2 has Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L 1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L 2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1- VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CL CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L.4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-V L1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL -Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH 2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL 1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or It has the structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has the structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL.リpeptideは、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VH1 L1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、 VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、It has a structure represented by VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1,VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH 2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-V H2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-C L, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL 2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc 、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2- VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3- VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-FcIn some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. has a structure represented by 2-L3-VH1-L4-CH1-L5-CL, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc,The structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc In some embodiments, the 1st polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, the 2nd polypeptide has the structure Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, V L1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-V H2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1 L1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-C L-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-V L1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL -Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1- L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1 -L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc has a structure. 2のpolypeptideは、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2 -VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc 、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、Structures represented by VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1,VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, V L1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、V L1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-FcIn some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. The first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-Fc, and the second polypeptide has structures such as Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc,VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、orVH1-L1-VH2-L It has a structure represented by 2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the 1st polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-Fc, and the 2nd polypeptide has a structure represented by Fc, VL1-VL 2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL1 VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-V H2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL 2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL 1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2 H2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、 VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL It has the structure represented by 2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the 1st polypeptide is VL1-VL2-VH2-V Having the structure represented by H1-CL-Fc, the second polypeptide is Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2 -VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1- VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-V.L2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3- VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2 -VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc 、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1- VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L 1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3 -VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc has the structure represented by In one aspect, the 1st polypeptide has the structure represented by VH1-VH2-VL2-VL1-CL-Fc, the 2nd polypeptide has the Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1,VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, V L1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、V L1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-FcIn some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CH1-CL-Fc, The second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc,It has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CH1-CL-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH I-Fc,It has a structure represented by VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-VL2-VH2-VH1-CL-CH1-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH2-VH1-CH1-CL, VH1-VH2-VL2-VL1-CH1-CL, VL1-VL2-VH2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1,VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4- CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1 -VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、V L1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3 -VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L 1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH It has a structure represented by 1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-VH2-VL2-VL1-CL-CH1-Fc. 、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1- VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH 2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1 -VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-V H2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-C H1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、V H1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4 -CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, and the second polypeptide has a structure represented by Fc, VL1-VL, 2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-V L2-L2-VH2-L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2- VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL 1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH 2-VH1-CL-CH1, VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH 1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1- VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2 -VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-F c、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2 -VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1- It has the structure represented by VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the 1st polypeptide is VH1-L1-VH2 -L2-VL2-L3-VL1-L4-CH1-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-VH2- L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、V L1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、 VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-C H1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH 2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL 2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5- CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L 5-CH1-Fc has a structure. In some embodiments, the 1st polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, and the 2nd polypeptide has a Fc、VL1-VL2-VH2-VH1、V H1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-Fc、VL1-L1-VL2-L2-VH2-L3 -VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1- VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、Structures represented by VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1, VL1-VL2-VH2-VH1-Fc,VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2- L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-V H2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2 -VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1 -L1-VL2-L2-VH2-L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4- CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1- CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2 -VH1-CH1-CL-Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1 -Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1- L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-FcIn some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, and the second polypeptide, は、Fc、VL1-VL2-VH2-VH1、VH1-VH2-VL2-VL1、VL1-L1-VL2-L2-VH2-L3-VH1 、VH1-L1-VH2-L2-VL2-L3-VL1、VL1-VL2-VH2-VH1-Fc、VH1-VH2-VL2-VL1-F c, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1- VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH 2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1 -VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1、VL1-L1-VL2-L2-VH2 -L3-VH1-L4-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL 1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1- VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL -Fc、VH1-VH2-VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH 2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1- It has the structure represented by VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the polypeptide 1 is VH1-L1-VH2- L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VH1, VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VH1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L2-VL H2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1-VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-C L、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1 1. CH1-L5-CL、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1、VL1-VL2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2 -VL2-VL1-CH1-CL-Fc、VL1-VL2-VH2-VH1-CL-CH1-Fc、VH1-VH2-VL2-VL1-CL-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc、VH1-L1-VH2-L2-VL2 -L3-VL1-L4-CH1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL -Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc、or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-C In some embodiments, the 1st polypeptide has the structure VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, the 2nd polypeptide has the structure Fc, VL1-VL2-VH2-VH 1. L3-VH1-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-Fc、VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc、VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc、VL1-VL2-VH2-VH1-CH1、VH1 -VH2-VL2-VL1-CH1、VL1-VL2-VH2-VH1-CL、VH1-VH2-VL2-VL1-CL、VL1-VL2-VH2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL2-VH2-VH1-CH1-Fc, VH1-VH2-VL2-VL1-CH1-Fc, VL1-VL2-VH2-VH1-CL-Fc, VH1-VH2-VL2-VL1-CL-Fc, VL1-VL2-VH2-VH1-CH1-CL-Fc, VH1-VH2-VL2-VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc. In some embodiments, the first polypeptide has a structure represented by VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc, and the second polypeptide is Fc, VL1-VL2-VH2-VH1, VH1-VH2-VL2-VL1, VL1-L1-VL2-L2-VH2-L3-VH1, VH1-L1-VH2-L2-VL2-L3-VL1,VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4- Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc, VL1-VL2-VH2-VH1-CH1, VH1-VH2-VL2-VL1-CH1, VL1-VL2-VH2-VH1-CL, VH1-VH2-VL2-VL1-CL, VL1-VL2-VH 2-VH1-CH1-CL、VH1-VH2-VL2-VL1-CH1-CL、VL1-VL2-VH2-VH1-CL-CH1、VH1-VH2-VL2-VL1-CL-CH1、VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1、VH1-L1-V H2-L2-VL2-L3-VL1-L4-CH1, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-C L, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1, VL1-VL 2-VH2-VH1-CH1-Fc、VH1-VH2-VL2-VL1-CH1-Fc、VL1-VL2-VH2-VH1-CL-Fc 、VH1-VH2-VL2-VL1-CL-Fc、VL1-VL2-VH2-VH1-CH1-CL-Fc、VH1-VH2-VL2- VL1-CH1-CL-Fc, VL1-VL2-VH2-VH1-CL-CH1-Fc, VH1-VH2-VL2-VL1-CL-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-Fc, VH1-L1-VH2-L2-VL2-L3- VL1-L4-CH1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH1-L5-CL-FcVH1-L1-VH2-L2-VL2-L3-VL1-L4-CH1-L5-CL-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CL-L5-CH1-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CL-L5-CH1-Fc.

[0091] In other aspects, the present invention is directed to an antigen-binding polypeptide comprising at least two Fc regions at its carboxy terminus, or an antigen-binding polypeptide complex comprising a polypeptide comprising at least two Fc regions at its carboxy terminus. The at least two Fc regions may be linked to the polypeptide via at least one amino acid linker. The at least two Fc regions may be linked to each other via at least one amino acid linker. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex may comprise a polypeptide having a structure represented by VL1-VL2-VH2-VH1 or VH1-VH2-VL2-VL1, which has two Fc regions. In some aspects, one or both Fc regions comprise the amino acid sequence of any one of SEQ ID NOs: 391-404, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 391-404. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex is selected from the group consisting of VL1-VL2-VH2-VH1-Fc-Fc, VH1-VH2-VL2-VL1-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VL1-L4-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-L5-Fc, or VH1-L1 -VH2-L2-VL2-L3-VL1-L4-Fc-L5-Fc, wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0092] In other aspects, the invention is directed to antigen-binding polypeptides comprising at least two CH3 regions, or antigen-binding polypeptide complexes comprising polypeptides comprising at least two CH3 regions. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex may comprise a polypeptide having a structure represented by VL1-VL2-VH2-VH1 or VH1-VH2-VL2-VL1, which has two CH3 regions. In some aspects, one or both CH3 regions comprise the amino acid sequence of any one of SEQ ID NOs: 416-419, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to any one of SEQ ID NOs: 416-419.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex is VL1-VL2-VH2-VH1-CH3, VH1-VH2-VL2-VL1-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-CH3, VL1-L1-VL2-L2-VH2-L3-VH1- L4-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH3, VL1-VL2-VH2-VH1-CH3-CH3, VH1-VH2-VL2-VL1-CH 3-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-CH3-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-CH3-CH3, VL1-L1-VL VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH3-CH3, VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH3-CH3, VL1-L1-VL2-L2-VH2-L3-VH1-L4-CH3-L5-CH3, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-CH3-L5-CH3. wherein VL1 is a first immunoglobulin light chain variable region, VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region, VH2 is a second immunoglobulin heavy chain variable region, CH3 is immunoglobulin heavy chain constant region 3, and L1, L2, L3, L4, and L5 are amino acid linkers.

[0093] Any one of the first polypeptides described herein can be combined with any one of the second and / or third polypeptides described herein to form an antigen-binding polypeptide complex of the invention.

[0094] All disclosures relating to antigen-binding polypeptide structures and antigen-binding polypeptide complex structures described herein apply to and can be combined with all VH and VL sequences described herein, including all target antigens described herein, and all VH and VL sequences and CDR sequences described herein.

[0095] In some embodiments, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes (e.g., antibodies or antigen-binding fragments thereof) that specifically bind to viral peptides, proteins, polypeptides, or fragments thereof. In some embodiments, the viral peptides, proteins, polypeptides, or fragments thereof include influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSV IgG1, RSV IgE, RSV IgE II, RSV IgE III, RSV IgE II ... G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hydodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyophyllae virusliyopneutiioniae), infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus A / Alaska 91 neuraminidase, equine influenza virus A / Miami 63 neuraminidase, equine influenza virus A / Kentucky 81 neuraminidase In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex is selected from the group consisting of equine influenza virus or equine herpesvirus antigens, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, bovine respiratory syncytial virus or bovine parainfluenza virus antigens, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, and human hepatitis C virus glycoproteins E1 and E2. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex is selected from the group consisting of influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, and human hepatitis C virus glycoproteins E1 and E2. In some embodiments, the antigen-binding polypeptide or polypeptide comprised within the antigen-binding complex specifically binds to at least one epitope on at least one viral protein selected from influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoproteins E1E2, or combinations thereof. For example, the polypeptide comprised within the antigen-binding polypeptide or antigen-binding complex can include VL1, VL2, VL3, VL4, VH1, VH2, VH3, and / or VH4 that specifically bind to a viral peptide, protein, polypeptide, or fragment thereof, such as influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, dengue virus matrix protein or other proteins, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, HIV-1 envelope glycoprotein, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneutyoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, The antibody may comprise a VL1 that specifically binds to an antigen of an equine influenza virus or equine herpesvirus, including equine influenza virus type A / Kentucky 81 neuraminidase, equine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D, an antigen of a bovine respiratory syncytial virus or bovine parainfluenza virus, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoproteins E1E2, or a combination thereof. For example, the antigen-binding polypeptide or polypeptide contained within the antigen-binding complex may be influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)For example, the polypeptide comprised within the antigen-binding polypeptide or antigen-binding polypeptide complex may comprise a VL2 that specifically binds to influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein ...G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoproteins E1 and E2, or a combination thereof. For example, the polypeptide comprised within the antigen-binding polypeptide or antigen-binding complex can include influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoproteins E1E2, or combinations thereof. For example, the polypeptide comprised within the antigen-binding polypeptide or antigen-binding complex can include influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSV G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV-1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, infectious gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneutyoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, neonatal calf diarrhea virusvirus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, mouse mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase, equine influenza virus or equine herpesvirus antigens including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, bovine respiratory syncytial virus or bovine parainfluenza virus antigens, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV G), For example, the polypeptides included within the antigen-binding polypeptide or antigen-binding complex may include a VH1 that specifically binds to influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein, RSV F glycoprotein, bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoprotein E1E2, or a combination thereof. G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195,Transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneutyoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G , infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus A / Alaska 91 neuraminidase, equine influenza virus A / Miami 63 neuraminidase, equine influenza virus A / Kentucky 81 neuraminidase The antigen-binding polypeptide or polypeptides comprised within the antigen-binding complex may include a VH2 that specifically binds to an antigen of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, an antigen of bovine respiratory syncytial virus or bovine parainfluenza virus, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine E virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, human hepatitis C virus glycoproteins E1 and E2, or a combination thereof. For example, the antigen-binding polypeptide or polypeptide comprised within the antigen-binding complex may include an influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral protein, RSV F glycoprotein,RSV G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuraminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, La Crosse virus glycoprotein, newborn calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D; antigens of bovine respiratory syncytial virus or bovine parainfluenza virus; bovine respiratory syncytial virus attachment protein (BRSV G);The antigen-binding polypeptides described herein or polypeptides of the antigen-binding polypeptide complexes described herein may comprise any combination of VL1, VL2, VL3, VL4, VH1, VH2, VH3, and / or VH4 that bind to a target described herein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of influenza virus neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a human respiratory syncytial virus (RSV) viral protein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a RSV F glycoprotein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a RSV G glycoprotein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a herpes simplex virus (HSV) viral protein.or fragment thereof. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of herpes simplex virus glycoprotein gB, gC, gD, or gE. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of chlamydia MOMP. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of PorB antigen. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of core protein, matrix protein, or other protein of dengue virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of measles virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of the simplex virus type 2 glycoprotein gB. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of the poliovirus 1 VPL. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of the envelope glycoprotein of HIV1. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of the hepatitis B surface antigen.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of diphtheria toxin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex The complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of the Streptococcus 24M epitope. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of gonorrhea pilin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of pseudorabies virus g50 (gpD). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of pseudorabies virus II (gpB). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of pseudorabies virus III (gpC). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of pseudorabies virus glycoprotein H. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of pseudorabies virus glycoprotein E. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of transmissible gastroenteritis glycoprotein 195. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of transmissible gastroenteritis matrix protein.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of porcine rotavirus glycoprotein 38. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of porcine parvovirus capsid protein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of Serpulina hyodysenteriae protective antigen. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine virus diarrhea glycoprotein 55. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of Newcastle disease virus hemagglutinin-neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of swine influenza hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of swine influenza neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of foot-and-mouth disease virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of classical swine fever virus.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of swine influenza virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of African swine fever virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of Mycoplasma lyopneutyoniae. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of Infectious Bovine Rhinotracheitis virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of glycoprotein E of infectious bovine rhinotracheitis virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of glycoprotein G. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody) or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of infectious laryngotracheitis virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of infectious laryngotracheitis virus glycoprotein G or glycoprotein I.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a La Crosse virus glycoprotein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a newborn calf diarrhea virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a Venezuelan equine encephalomyelitis virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a Punta Toro virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of a murine leukemia virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of mouse mammary tumor virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of hepatitis B virus core protein or hepatitis B virus surface antigen.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of equine influenza virus or equine herpesvirus, such as equine influenza virus A / Alaska 91 neuraminidase, equine influenza virus A / Miami 63 neuraminidase, equine influenza virus A / Kentucky 81 neuraminidase, equine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine respiratory syncytial virus or bovine parainfluenza virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine respiratory syncytial virus attachment protein (BRSV G). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine respiratory syncytial virus fusion protein (BRSV F). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine respiratory syncytial virus nucleocapsid protein (BRSVN). In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine parainfluenza virus type 3 fusion protein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of ovine parainfluenza virus type 3 hemagglutinin neuraminidase.In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of bovine E virus diarrhea virus glycoprotein 48 or glycoprotein 53. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of glycoprotein E of Dengue virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex (e.g., an antibody or antigen-binding fragment thereof) specifically binds to a viral peptide, protein, polypeptide, or fragment of glycoprotein E1E2 of human hepatitis C virus. Any of the antigen-binding polypeptide structures described herein and an antigen-binding polypeptide may be used to target one or more of the viral targets described herein. Any of the polypeptide complex structures can be used.

[0096] Sequences from antibodies or antibody fragments against known spike protein epitopes on any virus or overexpressed receptors on cancer cells can be inserted into the constructs disclosed herein to produce multispecific, multivalent polypeptides and polypeptide complexes that bind to epitopes on viruses or viral variants and to T cells that engage viruses or cancer cells. The Immune Epitope Database and Analysis Resource provides lists of epitope sequences associated with specific antigens and infectious organisms. Known VL / VH pairs and CDRs are selected for insertion into a plasmid or multiple plasmids that encode a fully functional multispecific, multivalent antibody. In a preferred embodiment, the source of the preferred initial antibody or monoclonal antibody is from a highly resistant subject that has developed broadly neutralizing antibodies that are resistant across evolving infectious viruses or cancer cells.

[0097] Viral antigens present in influenza A viruses include matrix protein 1, hemagglutinin, nucleoprotein RNA-directed RNA polymerase catalytic subunit, polymerase acidic protein, nuclear export protein, and polymerase basic protein 2. Epitope sequences include, for example, those selected from GILGFVFTL (SEQ ID NO: 422), PKYVKFQNTLKLAT (SEQ ID NO: 423), SRYWAIRTR (SEQ ID NO: 424), CTELKLSDY (SEQ ID NO: 425), ELRSRYWAI (SEQ ID NO: 426), ILRGSVAHK (SEQ ID NO: 427), VSDGGPNLY (SEQ ID NO: 428), FMYSEFHFI (SEQ ID NO: 429), AIMDKNIIL (SEQ ID NO: 430), NMLSTVLGV (SEQ ID NO: 431), FLKDVMESM (SEQ ID NO: 432), LPFEKSTVM (SEQ ID NO: 433), and FVRQCFNPM (SEQ ID NO: 434) disclosed in the above database.

[0098] Viral antigens present in influenza B virus are selected from the group consisting of nucleoprotein, hemagglutinin, nonstructural protein 1, neuraminidase, and matrix protein 1. Epitopes from such proteins are selected from, for example, KLGEFYNQMM (SEQ ID NO: 435), AVLLSNEGIINSEDE (SEQ ID NO: 436), AVLLSNEGIINSEDEH (SEQ ID NO: 437), AYDQSGRL (SEQ ID NO: 438), AYDQSGRLV (SEQ ID NO: 439), FPIMHDRTKI+OX(M4) (SEQ ID NO: 440), ITKNLNSLSELEVKN (SEQ ID NO: 441), ITKNLNSLSELEVKNLQ ​​(SEQ ID NO: 442), LAVLLSNEGIINSEDE (SEQ ID NO: 443), LAVLLSNEGIINSEDEH (SEQ ID NO: 444), and LPQSGRIVV (SEQ ID NO: 445) disclosed in the above database.

[0099] Antigens are selected from the group consisting of influenza viruses and surface glycoproteins: H5N1 influenza: H1N1: H1N2: H3N2: HA (hemagglutinin surface glycoprotein), NA (neuraminidase surface glycoprotein), H5, and H7. Others include respiratory syncytial virus (RSV). Antigens associated with RSV include protein M2-1, matrix protein, fusion glycoprotein F0, nucleoprotein, and small hydrophobic proteins. Epitopes present on these proteins include SYIGSINNI (SEQ ID NO: 446), NAITNAKII (SEQ ID NO: 447), KYKNAVTEL (SEQ ID NO: 448), NSELLSLINDMPITNDQKKLMSNN (SEQ ID NO: 449), NPKASLLSL (SEQ ID NO: 450), VYNTVISYI (SEQ ID NO: 451), TYMLTNSELL (SEQ ID NO: 452): WAICKRIPNKKPG (SEQ ID NO: 453), and KNRGIIKTFSN (SEQ ID NO: 454), among others.

[0100] Chlamydia. Antigens associated with Chlamydia trachomatis include the major outer membrane porin, serotype D; the chaperonin GroEL; an uncharacterized protein (UniProt: Q9Z7F3), a likely oxidoreductase CT_610, and integral membrane protein A. Epitope sequences include, for example, TLNPTI (SEQ ID NO: 455), ATLVVNRIRGGF (SEQ ID NO: 456), LNPTIA (SEQ ID NO: 457), SANNDAEIGNLI (SEQ ID NO: 458), PETISDPENRNKPSAE (SEQ ID NO: 459), AEGQLG (SEQ ID NO: 460), ARKLLLDNL (SEQ ID NO: 461), ASFVNPIYL (SEQ ID NO: 462), DVVDGMNFNRGY (SEQ ID NO: 463), NMFTPYIGV (SEQ ID NO: 464), and NLVGLIGVKGSSIAADQLPNVGIT (SEQ ID NO: 465).

[0101] Adenoviridae. Antigens associated with human adenovirus C include early E1A protein, hexon protein, DNA-binding protein, E1B 55 kDa protein, and DNA polymerase. Epitope sequences include, for example, SGPSNTPPEI (SEQ ID NO: 466), TDLGQNLLY (SEQ ID NO: 467), LTDLGQNLLY (SEQ ID NO: 468), FALSNAEDL (SEQ ID NO: 469), DEPTLLYVLFEVFDV (SEQ ID NO: 470), KYSPSNVKI (SEQ ID NO: 471), MPNRNYIAF (SEQ ID NO: 472), VDCYINLGARWSLDY (SEQ ID NO: 473), VNIRNCCYI (SEQ ID NO: 474), RNFQPMSRQVVDDTKYKDYQQVGILHQHNN (SEQ ID NO: 475), LPKLTPFAL (SEQ ID NO: 476), and FQRPTISSNSHAIFR (SEQ ID NO: 477).

[0102] Mastadenovirus. Human mastadenovirus C has various antigens related to viral infection. These include the early E1A protein, hexon protein, DNA-binding protein, E1B 55 kDa protein, DNA polymerase, and fiber protein. Epitope sequences include, for example, SGPSNTPPEI (SEQ ID NO: 478), TDLGQNLLY (SEQ ID NO: 479), LTDLGQNLLY (SEQ ID NO: 480), FALSNAEDL (SEQ ID NO: 481), DEPTLLYVLFEVFDV (SEQ ID NO: 482), KYSPSNVKI (SEQ ID NO: 483), MPNRPNYIAF (SEQ ID NO: 484), VDCYINLGARWSLDY (SEQ ID NO: 485), LPKLTPFAL (SEQ ID NO: 486), FQRPTISSNSHAIFR (SEQ ID NO: 487), and GKYTTETFATNSYTPSYIAQE (SEQ ID NO: 488).

[0103] Avian adenovirus. Poultry adenovirus C has a hexon protein as one of its antigens, including the epitopes DYDDYNIGTT (SEQ ID NO: 489), KISGVFPNP (SEQ ID NO: 490), PLAPKESMFN (SEQ ID NO: 491), and ETLIEDDVSGQGKELGVNLNPAGPITADEQGL (SEQ ID NO: 492).

[0104] Herpesviridae. Antigens depend on the specific organism, and human herpesvirus 5 (human cytomegalovirus) and human herpesvirus 4 (Epstein-Barr virus) include, for example, NLVPMVATV (SEQ ID NO: 493), GLCTLVAML (SEQ ID NO: 494), TPRVTGGGAM (SEQ ID NO: 495), SSIEFARL (SEQ ID NO: 496), CLGGLLTMV (SEQ ID NO: 497), FLRGRAYGL (SEQ ID NO: 498), TPHFMPTNL (SEQ ID NO: 499), RAKFKQLL (SEQ ID NO: 500), RPPIFIRRL (SEQ ID NO: 501), VLEETSVML (SEQ ID NO: 502), IVTDFSVIK (SEQ ID NO: 503), IPSINVHHY (SEQ ID NO: 504), QYDPVAALF (SEQ ID NO: 505), HPVGEADYFEY (SEQ ID NO: 506), VTEHDTLLY (SEQ ID NO: 507), HGIRNASFI (SEQ ID NO: 508), AVFDRKSDAK (SEQ ID NO: 509), TPLHEQHGM (SEQ ID NO: 510), YSEHPTFTSQY (SEQ ID NO: 511), YVLDHLIVV (SEQ ID NO: 512), YLLEMLWRL (SEQ ID NO: 513), FLYALALLL (SEQ ID NO: 514), QAKWRLQTL (SEQ ID NO: 515), ELRRKMMYM (SEQ ID NO: 516), and RPHERNGFTVL (SEQ ID NO: 517), and the like. processivity factor), ribonucleoside-diphosphate reductase large subunit-like protein, replication and transcription activators, and latent infection membrane proteins.

[0105] Herpes simplex virus 1 (human herpesvirus 1). Antigens include envelope glycoprotein B, ribonucleoside-diphosphate reductase large subunit, envelope glycoprotein D, tegument protein UL46, mRNA export factor, capsid apex component 2, and ribonucleoside-diphosphate reductase small subunit. Epitope sequences include SSIEFARL (sequence number 518), QTFDRGRL (sequence number 519), SLKMADPNRFRGKDLP (sequence number 520), QPPSLPITVYYAVLERACTSVLLNAPSEAPQIVR (sequence number 521), RLNELLAYV (sequence number 522), RMLGDVMAV (sequence number 523), KYALADASLKMADPNRFRGKDLP (sequence number 524), SLPITVTTA (sequence number 525), DPEDSALL (sequence number 526), ​​and DYATLGVGV (sequence number 527).

[0106] Herpes simplex virus 2 (human herpesvirus 2). Antigens include, for example, RPRGEVRFL (SEQ ID NO: 528), SSIEFARL (SEQ ID NO: 529), EEVDMTPADALDDFD (SEQ ID NO: 530), GLADTVVAC (SEQ ID NO: 531), ASDSLNNEY (SEQ ID NO: 532), DFEFEQMFTDAMG (SEQ ID NO: 533), EVDMTPADAL (SEQ ID NO: 534), PEEFEGAGDGEPPEDDDS (SEQ ID NO: 535), FLWEDQTLL (SEQ ID NO: 536), FLVDAIVRVA (SEQ ID NO: 537), GPADAPPGSPAPPPPEHRGG (SEQ ID NO: 538), GPHETITAL (SEQ ID NO: 539), KYALADPSLKMADPNRFRGKNLP (SEQ ID NO: 540), NNY GSTIEGLL (SEQ ID NO: 541), PEEFEGAGDGEPPEDDDSAT (SEQ ID NO: 542), PPLYATGRLSQAQLMPSPPM (SEQ ID NO: 543), TQPELVPEDPED (SEQ ID NO: 544), YTSTLLPPELSDTTN (SEQ ID NO: 545), DPSLKMADPNRFRGKNLPVL (SEQ ID NO: 546), PELVPEDPEDSALLEDPAGT (SEQ ID NO: 547), HGPSLYRTF (SEQ ID NO: 548), NKRVFCAAVGRLA (SEQ ID NO: 549), PMRARPRGEVRFL (SEQ ID NO: 550), VFCAAVGRL (SEQ ID NO: 551), and LGNRLCGPATAAWAG (SEQ ID NO: 552), and Immune Further disclosed in the Epitope Database are tegument protein VP22, envelope glycoprotein B, tegument protein VP16, tegument protein UL47, tegument protein UL46, tegument protein VP16, envelope glycoprotein G, capsid vertex component 2, capsid scaffolding protein, envelope glycoprotein D, mRNA export factor, major viral transcription factor ICP4 homolog, with antigenic epitopes selected from those further disclosed in the Epitope Database.

[0107] Herpes simplex virus 5 (human herpesvirus 5). Antigens include the 65 kDa phosphoprotein, immediate early protein IE1, envelope glycoprotein H, other human herpesvirus 5 proteins, and envelope glycoprotein B. Epitope sequences include NLVPMVATV (SEQ ID NO: 553), TMYGGISLL (SEQ ID NO: 554), VLEETSVML (SEQ ID NO: 555), LDPHAFHLLL (SEQ ID NO: 556), RIFAELEGV (SEQ ID NO: 557), RPHERNGFTVL (SEQ ID NO: 558), VFPTKDVAL (SEQ ID NO: 559), VLAELVKQI (SEQ ID NO: 560), VLPHETRLL (SEQ ID NO: 561), KRLDVCRAKMGYM (SEQ ID NO: 562), GGGAMAGASTSAGRKRKS (SEQ ID NO: 563), AALFFFDID (SEQ ID NO: 564), AGILARNLVPMVATV (SEQ ID NO: 565), ALFFFDIDLL (SEQ ID NO: 566), ANETIYNTTLKYGDV (SEQ ID NO: 567), ARAKKDELRRKMMYM (SEQ ID NO: 568), ARNLVPMVATVQGQN (SEQ ID NO: 569), ASTAAPPYTNEQAYQMLLAL (SEQ ID NO: 570), AVGGAVASV (SEQ ID NO: 571), DEEEAIVAYT (SEQ ID NO: 572), DEEEAIVAYTL (SEQ ID NO: 573), DPVAALFFF (SEQ ID NO: 574), EEAIVAYTL (SEQ ID NO: 575), EECQLPSLKIFIAGNSAY (SEQ ID NO: 576), or EEEAIVAYTL (SEQ ID NO: 577), and others disclosed in public databases such as the Immune Epitope Database.

[0108] Other antigens include herpes simplex virus 6, Leviviridae, Levivirus, Enterobacteriaceae MS2, Allorevirus, Poxviridae, Chordopoxvirinae (cowpox virus or vaccinia virus); antigens include CPXV202 protein, intermediate transcription factor 3 small subunit, putative nuclease G5, interferon antagonist C7, protein A47, major core protein 4b, DNA-directed RNA polymerase 147 kDa polypeptide, mRNA capping enzyme regulatory subunit, envelope protein H3, protein B6, telomere binding protein I1, protein K3, poxin, protein A19, assembly protein G7, protein F12, protein A46, protein A6, DNA polymerase, profiling, RNA-binding protein E3, and serine protease inhibitor 1. Antigens include TSYKFESV (SEQ ID NO: 578), ITYRFYLI (SEQ ID NO: 579), ILDDNLYKV (SEQ ID NO: 580), KVDDTFYYV (SEQ ID NO: 581), AAFEFINSL (SEQ ID NO: 582), KSYNYMLL (SEQ ID NO: 583), MPAYIRNTL (SEQ ID NO: 584), RVYEALYYV (SEQ ID NO: 585), IGMFNLTFI (SEQ ID NO: 586), SLSAYIIRV (SEQ ID NO: 587), LMYDIINSV (SEQ ID NO: 588), RLYDYFTRV (SEQ ID NO: 589), YSLPNAGDVI (SEQ ID NO: 590), Y It has an epitope sequence selected from SQVNKRYI (SEQ ID NO: 591), VSLDYINTM (SEQ ID NO: 592), TLPEVISTI (SEQ ID NO: 593), FLTSVINRV (SEQ ID NO: 594), GFFDFVNFV (SEQ ID NO: 595), VLYDEFVTI (SEQ ID NO: 596), FPYEGGKVF (SEQ ID NO: 597), LMDENTYAM (SEQ ID NO: 598), NLFDIPLLTV (SEQ ID NO: 599), VGPSNSPTF (SEQ ID NO: 600), YAPVSPIVI (SEQ ID NO: 601), and HVDGKILFV (SEQ ID NO: 602), etc.

[0109] Parapoxvirus (orf virus). Antigens include selectable proteins having the epitope sequences AAFEFRDL (SEQ ID NO: 603), AIIKYTDL (SEQ ID NO: 604), AIYAFRLT (SEQ ID NO: 605), AIYGFGVTF (SEQ ID NO: 606), ANVDFMEYV (SEQ ID NO: 607), and EQFSFSNV (SEQ ID NO: 608), ORF011 putative EEV envelope phospholipase, ORF052 putative IMV membrane protein, ORF110 EEV glycoprotein, ORF094 putative phosphorylated IMV membrane protein, ORF056 RNA polymerase subunit RPO147, and ORF101 RNA polymerase subunit RPO132. Other antigens that bind epitopes, as well as associated antibodies and antibody fragments thereof, include avian poxvirus, capripoxvirus, leporipoxvirus, suipoxvirus, molluskipoxvirus, Entomopoxvirinae, Papovaviridae, polyomavirus, papillomavirus, Paramyxoviridae, paramyxovirus, parainfluenza virus type 1, morbillivirus, measles virus, rubulavirus, mumps virus, pneumovirinae, pneumovirus, metapneumovirus, avian pneumovirus, human metapneumovirus, picornaviridae, and enteroviruses (enterovirus A, coxsackievirus A).Antigens include TYTFGEHKQEKDLEY (SEQ ID NO: 609), TEDSHPPYKQTQPGA (SEQ ID NO: 610), PESRESLAWQTATNP (SEQ ID NO: 611), FGEHKQEKDL (SEQ ID NO: 612), AGGTGTEDSHPPYKQ (SEQ ID NO: 613), FGEHKQEKDL (SEQ ID NO: 614), AGGTGTEDSHPPYKQ (SEQ ID NO: 615), FGEHKQEKDLEYGAC (SEQ ID NO: 616), HYRAHARDGVFDYYT (SEQ ID NO: 617), and TYTFGEHKQEKDLEY (SEQ ID NO: 618). No. 617), KQEDK (SEQ ID NO: 618), GDPIADMIDQTVNNQ (SEQ ID NO: 619), YPTFGEHLQANDLDY (SEQ ID NO: 620), LEGTTNPNT (SEQ ID NO: 621), VSSHRLDDTGEVPALQ (SEQ ID NO: 622), RIYMRMKHVR (SEQ ID NO: 623), TSKSKYPLVV (SEQ ID NO: 624), and DGYPTFGEHKQEKDL (SEQ ID NO: 625).

[0110] Rhinoviruses and Hepatoviruses. Human hepatitis A virus (hepatovirus A) having a genomic polyprotein as antigen with epitopes YMYAVSGAL (SEQ ID NO: 626), FWRGDLVFDFQV (SEQ ID NO: 627), MNMSKQGIFQTVGSGLDHILSLA (SEQ ID NO: 628), TVSTEQNVPDPQVGI (SEQ ID NO: 629), ASICQMFCFWRGDLVFDFQV (SEQ ID NO: 630), DHMSIYKFMGRSHFLCTFTF (SEQ ID NO: 631), FPELKPGESTHTSDHMSIYK (SEQ ID NO: 632), additionally as disclosed in IED. Others include cardioviruses, andaptoviruses, Reoviridae, orthoreoviruses, orbiviruses, rotaviruses, cypoviruses, phyziviruses, phytreoviruses, oryzaviruses, Retroviridae, mammalian type B retroviruses, mammalian type C retroviruses, avian type C retroviruses, type D retrovirus group, BLV-HTLV retroviruses, lentiviruses, human immunodeficiency virus type 1, human immunodeficiency virus type 2, HTLV-I and -II viruses, herpes simplex virus, Epstein-Barr virus, cytomegalovirus, hepatitis viruses (HCV, HAV, HBV, HDV, HEV), Toxoplasma gondii virus, Treponema pallidum pallidium virus, human T-lymphotropic virus, encephalitis virus, West Nile virus, dengue virus, varicella-zoster virus, measles, mumps, rubella, spumavirus, Flaviviridae, Hepatitis C virus, Hepadnaviridae, Hepatitis B virus, Togaviridae, Alphavirus, Sindbis virus, Rubivirus, Rubella virus, Rhabdoviridae, Vesiculovirus, Lyssavirus, Ephemelovirus, Cytorhabdovirus, Nucleorhabdovirus, Arenaviridae, Arenavirus, Lymphocytic choriomeningitis virus, Yippi virus, Lassa virus, and Torovirus.

[0111] Thus, the multispecific and multivalent antibody constructs provided herein have embedded sequences that target and bind to epitopes on viral peptides, proteins, polypeptides, or glycosylated versions thereof in a subject in need thereof, including influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSV IgG1, RSV IgE ... G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus Glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, govine bovine virus diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuroaminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma liyopneutiinniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein e, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, Las CrossCross) virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase, equine influenza virus or equine herpesvirus antigens including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, bovine respiratory syncytial virus or bovine parainfluenza virus antigens, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV G), F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine viral diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, and human hepatitis C virus glycoproteins E1E2. All antigen-binding polypeptide structures and all antigen-binding polypeptide complex structures described herein may be selected from the group consisting of one or more of the viral antigen targets described herein, i.e., influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSVG glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine virus diarrhea glycoprotein 55 , Newcastle disease virus hemagglutinin-neuroaminidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein e, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, Las Crosses virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)In some embodiments, the antigen-binding polypeptides or antigen-binding polypeptide complexes described herein may specifically bind to one or more (two or more, three or more, or four) of influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein ...G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epidermal growth factor receptor (EGFR) Tope, Neisseria gonorrhoeae pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle Disease virus hemagglutinin-neuroinidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, Las Crosses virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase Antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, antigens of bovine respiratory syncytial virus or bovine parainfluenza virus, and bovine respiratory syncytial virus attachment protein (BRSV)In some embodiments, the antigen-binding polypeptides or antigen-binding polypeptide complexes described herein comprise a VL1 that specifically binds to influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein ...G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydial MOMP and PorB antigens, core protein, dengue virus matrix protein or other proteins, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, HIV-1 envelope glycoprotein, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysene terrier protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuroinidase, swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, glycoprotein of Las Crosses virus, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus type A / Alaska 91 neuraminidase , equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase, equine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D, equine influenza virus or equine herpesvirus antigens including bovine respiratory syncytial virus or bovine parainfluenza virus antigens, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine e virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, or human hepatitis C virus glycoproteins E1 and E2. In some embodiments, the antigen-binding polypeptides described herein or antigen-binding polypeptide complexes described herein are selected from the group consisting of influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSV G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydia MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus, measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, envelope glycoprotein of HIV1, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudomyxin, leukemia virus ... Rabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuroinidase,Swine influenza hemagglutinin, swine influenza neuraminidase, foot-and-mouth disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, Las Crosses virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or a fragment or derivative thereof, equine influenza virus A / Alaska 91 neuraminidase, equine influenza virus A / Miami 63 neuraminidase, equine influenza virus A / Kentucky 81 neuraminidase The antibody comprises a VH1 that specifically binds to an antigen of an equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D, an antigen of a bovine respiratory syncytial virus or bovine parainfluenza virus, bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase, bovine e virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, or human hepatitis C virus glycoproteins E1 and E2. In some embodiments, the antigen-binding polypeptides described herein or antigen-binding polypeptide complexes described herein are selected from the group consisting of influenza virus neuraminidase, influenza virus hemagglutinin, human respiratory syncytial virus (RSV) viral proteins, RSV F glycoprotein, RSV G glycoprotein, herpes simplex virus (HSV) viral proteins, herpes simplex virus glycoproteins gB, gC, gD, and gE, chlamydia MOMP and PorB antigens, core protein, matrix protein or other proteins of dengue virus,Measles virus hemagglutinin, herpes simplex virus type 2 glycoprotein gB, poliovirus type 1 VP1, HIV-1 envelope glycoprotein, hepatitis B surface antigen, diphtheria toxin, streptococcal 24M epitope, gonococcal pilin, pseudorabies virus g50 (gpD), pseudorabies virus II (gpB), pseudorabies virus III (gpC), pseudorabies virus glycoprotein H, pseudorabies virus glycoprotein E, transmissible gastroenteritis glycoprotein 195, transmissible gastroenteritis matrix protein, porcine rotavirus glycoprotein 38, porcine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, bovine viral diarrhea glycoprotein 55, Newcastle disease virus hemagglutinin-neuroinidase, swine influenza hemagglutinin, swine influenza neuraminidase, oral Hoof disease virus, hog cholera virus, swine influenza virus, African swine fever virus, Mycoplasma lyopneumoniae, infectious bovine rhinotracheitis virus, infectious bovine rhinotracheitis virus glycoprotein E, glycoprotein G, infectious laryngotracheitis virus, infectious laryngotracheitis virus glycoprotein G or glycoprotein I, Las Crosses virus glycoprotein, neonatal calf diarrhea virus, Venezuelan equine encephalomyelitis virus, Punta Toro virus, murine leukemia virus, murine mammary tumor virus, hepatitis B virus core protein and hepatitis B virus surface antigen or fragments or derivatives thereof, equine influenza virus type A / Alaska 91 neuraminidase, equine influenza virus type A / Miami 63 neuraminidase, equine influenza virus type A / Kentucky 81 neuraminidase antigens of equine influenza virus or equine herpesvirus, including equine herpesvirus type 1 glycoprotein B and equine herpesvirus type 1 glycoprotein D; antigens of bovine respiratory syncytial virus or bovine parainfluenza virus; bovine respiratory syncytial virus attachment protein (BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F), bovine respiratory syncytial virus nucleocapsid protein (BRSVN), bovine parainfluenza virus type 3 fusion protein, bovine parainfluenza virus type 3 hemagglutinin neuraminidase;and VH2 that specifically binds to bovine e virus diarrhea virus glycoprotein 48 and glycoprotein 53, dengue virus glycoprotein E, or human hepatitis C virus glycoproteins E1E2. The antigen-binding polypeptides and antigen-binding polypeptide complexes described herein can contain any combination of VH1, VH2, VL1, and VL2 that binds to the targets described herein. For example, the viral peptide, protein, polypeptide, or glycosylated version thereof is selected from the group consisting of influenza virus neuraminidase, influenza virus hemagglutinin, herpes simplex virus (HSV) viral protein, core protein, matrix protein, or other proteins of dengue virus and swine influenza virus proteins. All antigen-binding polypeptide structures described herein and all antigen-binding polypeptide complex structures described herein may specifically bind to one or more of the viral antigen targets described herein, i.e., one or more (such as two or more, three or more, or four) of influenza virus neuraminidase, influenza virus hemagglutinin, herpes simplex virus (HSV) viral proteins, core protein, matrix protein, or other proteins of dengue virus and swine influenza virus proteins. In some embodiments, the antigen-binding polypeptides and antigen-binding polypeptide complexes may specifically bind to influenza virus neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL1 that specifically binds to influenza virus neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL2 that specifically binds to influenza virus neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH1 that specifically binds to influenza virus neuraminidase. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH2 that specifically binds to influenza virus neuraminidase.The antigen-binding polypeptides and antigen-binding polypeptide complexes may specifically bind to influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL1 that specifically binds to influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL2 that specifically binds to influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH1 that specifically binds to influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH2 that specifically binds to influenza virus hemagglutinin. In some embodiments, the antigen-binding polypeptide and antigen-binding polypeptide complexes may specifically bind to herpes simplex virus (HSV) viral proteins. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL1 that specifically binds to herpes simplex virus (HSV) viral proteins. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL2 that specifically binds to herpes simplex virus (HSV) viral proteins. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH1 that specifically binds to a herpes simplex virus (HSV) viral protein. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH2 that specifically binds to a herpes simplex virus (HSV) viral protein. In some embodiments, the antigen-binding polypeptide and antigen-binding polypeptide complex may specifically bind to HIV. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL1 that specifically binds to dengue virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VL2 that specifically binds to dengue virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex comprises a VH1 that specifically binds to dengue virus. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex isIn some embodiments, the antigen-binding polypeptides and antigen-binding polypeptide complexes described herein comprise a VL1 that specifically binds to swine influenza virus. In some embodiments, the antigen-binding polypeptides described herein or the antigen-binding polypeptide complexes described herein comprise a VL1 that specifically binds to swine influenza virus. In some embodiments, the antigen-binding polypeptides described herein The antigen-binding polypeptides described herein or antigen-binding polypeptide complexes described herein comprise a VL2 that specifically binds to swine influenza virus. In some embodiments, the antigen-binding polypeptides described herein or antigen-binding polypeptide complexes described herein comprise a VH1 that specifically binds to swine influenza virus. In some embodiments, the antigen-binding polypeptides described herein or antigen-binding polypeptide complexes described herein comprise a VH2 that specifically binds to swine influenza virus.

[0112] A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H 6、B7H7、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL4、CCL5、CCL 7、CCL8、CCL11、CCL15、CCL17、CCL19、CCL20、CCL21、CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30 CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123 CD133, CD137, CD137L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL2 、CXCL13、CXCR3、cMet、CTLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、 EGFR、ENTPD1、EpCAM、FCER1、FCER1A、FCER2、FGFR、FLAP、FOLH 1、Gi24、GITR、GITRL、GPR5、GP100、GPRC5D、HER2、HER3、ICOSL ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2R, I L1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7 IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, IL13, IL13Ral, I L13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL 33、IL35、ITGB4、ITK、KIR、LAG3、LAMP1、レプチ、LPFS2、MHC、、、I I、MUC-1、MUC-16、NCR3LG1、NKG2D、NKp46、NTPDase-1、OX40、 OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP, SISP1, SL C, SPG64, ST2, STEAP1, STEAP2, Syk, STEAP1, TROP2, TACI TDO, TGF, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9The present invention relates to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to TMEF1, TNFα, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM, or a combination thereof. In some embodiments, the antigen-binding polypeptide or antigen-binding polypeptide complex is selected from the group consisting of A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, and CCL25. CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, C D70, CD80, CD86, CD123, CD133, CD137, CD137L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF- 1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4, DNGR-1, E- Cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP 100, GPRC5D, HER2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2Rbeta, IL1, IL IA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, IL13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40,Specifically binds to at least one epitope on at least one antigen selected from the group consisting of OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM. In some embodiments, the antigen-binding polypeptide or polypeptide comprised within the antigen-binding complex is selected from the group consisting of A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123, CD133, CD137, CD137L, CD160 , CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4, DNGR-1, E-cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP100, GPRC5D, HER2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, I DO, IFNa, IgE, IGF1R, IL2Rbeta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, IL13, IL13Ral,IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC -1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, The antibodies may include VL1, VL2, VL3, VL4, VH1, VH2, VH3, and / or VH4 that specifically bind to STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFα, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, the antigen-binding polypeptide or polypeptide contained within the antigen-binding complex may be A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD 80, CD86, CD123, CD133, CD137, CD137L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3 , CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4, DNGR-1, E-cadherin, EGFR, ENTPD1, ​​Ep CAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP100, GPRC5D, HER2, HER3, ICOSL,ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2Rbeta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12 , IL13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG 1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, the antigen-binding polypeptide or polypeptide contained within the antigen-binding complex may be A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123, CD133, CD137, CD13 7L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4,DNGR-1、E-カドヘリン、EGFR、ENTPD1、EpCAM、FCER1、FCER1A、FCER2、FGFR、FLAP、FOLH1、Gi24、GITR、GITRL、GPR5、GP100、GPRC5D、HER2、HER3、ICOSL、IC、 OS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2R beta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, I L13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG1 , NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, the antigen-binding polypeptide or polypeptide contained within the antigen-binding complex may be A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123, CD133, CD137, CD13 7L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4,DNGR-1, E-cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP100, GPRC5D, HER2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2R beta , IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO , IL12, IL13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4 , ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, the antigen-binding polypeptide or polypeptide contained within the antigen-binding complex may be A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25 CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123, CD133, CD137, CD137L,CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cM et, CTLA4, DLL3, DLL4, DNGR-1, E-cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, G ITRL, GPR5, GP100, GPRC5D, HER2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2R beta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, IL13, IL13Ral, IL13Ra 2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, ITGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II , MUC-1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, The VL4 may include a VL4 that specifically binds to SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFa, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, the antigen-binding polypeptide or polypeptide comprised within the antigen-binding complex can be A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, CCL25, CCR3,CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD86, CD123, CD133, CD137, CD1 37L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DL L3, DLL4, DNGR-1, E-cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP100, GPRC5D, HE R2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2Rbeta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, I L7, IL7Ra, IL8, IL9, IL9R, IL10, rhILlO, IL12, IL13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, I TGB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD- For example, the antigen-binding polypeptide or polypeptide comprised within the antigen-binding complex may include a VH1 that specifically binds to L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2, TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFα, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM.B7H3、B7H4、B7H5、B7H6、B7H7、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL4 、CCL5、CCL7、CCL8、CCL11、CCL15、CCL17、CCL19、CCL20、CCL21、CCL25 CCR3、CCR4、CD3、CD16A、CD19、CD20、CD24、CD27、CD28、CD30、CD38 、CD39、CD40、CD40L、CD47、CD52、CD70、CD80、CD86、CD123、CD133、C D137、CD137L、CD160、CD272、CEACAM5、CLEC9、CLEC91、CRTH2、CSF -1、CSF-2、CSF-3、CXCL1、CXCL2、CXCL4、CXCL12、CXCL13、CXCR3、cM et, CTLA4, DLL3, DLL4, DNGR-1, E-CARDHON, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, G P100、GPRC5D、HER2、HER3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、IDO、I FNa、IgE、IGF1R、IL2Rベータ、IL1、ILIA、IL1B、IL1F10、IL2、IL4、IL4R a、IL5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、IL9R、IL10、rhILlO、IL12 、IL13、IL13Ral、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、I L25、IL7、IL33、IL35、ITGB4、ITK、KIR、LAG3、LAMP1、レプチン、LPFS2、M HCクラスII、MUC-1、MUC-16、NCR3LG1、NKG2D、NKp46、NTPDase-1、OX40 、OX40L、PD-1、PD-L1、PD-L2、PROM1、S152、SIRPアルファ、SISP1、SLC、 SPG64、ST2、STEAP1、STEAP2、Sykキナーゼ、STEAP1、TROP2、TACI、TDO、T GFBETA、T14、TIGIT、TIM3、TLR、TLR2、TLR4、TLR5、TLR9、TMEF1、TNF a、TNFRSF7、Tp55、TREMl、TSLP、TSLPR、TWEAK、VEGF、VISTA、Vstm3、For example, the polypeptide contained in the antigen-binding polypeptide or antigen-binding complex may be selected from the group consisting of A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7, H4、B7H5、B7H6、B7H7、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL4、CCL 5、CCL7、CCL8、CCL11、CCL15、CCL17、CCL19、CCL20、CCL21、CCL25 CCR3、CCR4、CD3、CD16A、CD19、CD20、CD24、CD27、CD28、CD30、CD38 、CD39、CD40、CD40L、CD47、CD52、CD70、CD80、CD86、CD123、CD133、C D137、CD137L、CD160、CD272、CEACAM5、CLEC9、CLEC91、CRTH2、CSF -1、CSF-2、CSF-3、CXCL1、CXCL2、CXCL4、CXCL12、CXCL13、CXCR3、cM et, CTLA4, DLL3, DLL4, DNGR-1, E-CARDHON, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A, FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, G P100、GPRC5D、HER2、HER3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、IDO、I FNa、IgE、IGF1R、IL2Rベータ、IL1、ILIA、IL1B、IL1F10、IL2、IL4、IL4R a、IL5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、IL9R、IL10、rhILlO、IL12 、IL13、IL13Ral、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、I L25、IL7、IL33、IL35、ITGB4、ITK、KIR、LAG3、LAMP1、レプチン、LPFS2、M HCクラスII、MUC-1、MUC-16、NCR3LG1、NKG2D、NKp46、NTPDase-1、OX40 、OX40L、PD-1、PD-L1、PD-L2、PROM1、S152、SIRPアルファ、SISP1、SLC、 SPG64、ST2、STEAP1、STEAP2、Sykキナーゼ、STEAP1、TROP2、TACI、TDO、T GFBETA、T14、TIGIT、TIM3、TLR、TLR2、TLR4、TLR5、TLR9、TMEF1、TNF a、TNFRSF7、Tp55、TREMl、TSLP、TSLPR、TWEAK、VEGF、VISTA、Vstm3、For example, the polypeptide contained within the antigen-binding polypeptide or antigen-binding complex may be selected from the group consisting of A2AR, APRIL, ATPDase, BAFF, BAFFR, BCMA, BlyS, BTK, BTLA, B7DC, B7H1, B7H2, B7H3, B7H4, B7H5, B7H6, B7H7, B7RP1, B7-4, C3, C5, CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL15, CCL17, CCL19, CCL20, CCL21, and CCL25. CCR3, CCR4, CD3, CD16A, CD19, CD20, CD24, CD27, CD28, CD30, CD38, CD39, CD40, CD40L, CD47, CD52, CD70, CD80, CD8 6, CD123, CD133, CD137, CD137L, CD160, CD272, CEACAM5, CLEC9, CLEC91, CRTH2, CSF-1, CSF-2, CSF-3, CXCL1, CXCL 2, CXCL4, CXCL12, CXCL13, CXCR3, cMet, CTLA4, DLL3, DLL4, DNGR-1, E-cadherin, EGFR, ENTPD1, ​​EpCAM, FCER1, FCER1A , FCER2, FGFR, FLAP, FOLH1, Gi24, GITR, GITRL, GPR5, GP100, GPRC5D, HER2, HER3, ICOSL, ICOS, HHLA2, HMGB1, HVEM, IDO, IFNa, IgE, IGF1R, IL2Rbeta, IL1, ILIA, IL1B, IL1F10, IL2, IL4, IL4Ra, IL5, IL5R, IL6, IL7, IL7Ra, IL8, IL9, I L9R, IL10, rhILlO, IL12, IL13, IL13Ral, IL13Ra2, IL15, IL17, IL17Rb, IL18, IL22, IL23, IL25, IL7, IL33, IL35, IT GB4, ITK, KIR, LAG3, LAMP1, leptin, LPFS2, MHC class II, MUC-1, MUC-16, NCR3LG1, NKG2D, NKp46, NTPDase-1, OX40, OX40L, PD-1, PD-L1, PD-L2, PROM1, S152, SIRP alpha, SISP1, SLC, SPG64, ST2, STEAP1, STEAP2, Syk kinase, STEAP1, TROP2,The VL1 may include a VH4 that specifically binds to TACI, TDO, TGFBETA, T14, TIGIT, TIM3, TLR, TLR2, TLR4, TLR5, TLR9, TMEF1, TNFα, TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, or WUCAM. For example, VL1 may specifically bind to CD3. For example, VL1 may specifically bind to CD19. For example, VL1 may specifically bind to HER2. For example, VL1 may specifically bind to CD20. For example, VL1 may specifically bind to CD28. For example, VL1 may specifically bind to CD38. For example, VL1 may specifically bind to Trop2. For example, VL1 may specifically bind to cMet. For example, VL2 may specifically bind to CD3. For example, VL2 may specifically bind to CD19. For example, VL2 may specifically bind to HER2. For example, VL2 may specifically bind to CD20. For example, VL2 may specifically bind to CD28. For example, VL2 may specifically bind to CD38. For example, VL2 may specifically bind to Trop2. For example, VL2 may specifically bind to cMet. For example, VL3 may specifically bind to CD3. For example, VL3 may specifically bind to CD19. For example, VL3 may specifically bind to HER2. For example, VL3 may specifically bind to CD20. For example, VL3 may specifically bind to CD28. For example, VL3 may specifically bind to CD38. For example, VL3 may specifically bind to Trop2. For example, VL3 may specifically bind to cMet. For example, VL4 may specifically bind to CD3. For example, VL4 may specifically bind to CD19. For example, VL4 may specifically bind to HER2. For example, VL4 may specifically bind to CD20. For example, VL4 may specifically bind to CD28. For example, VL4 may specifically bind to CD38. For example, VL4 may specifically bind to Trop2. For example, VL4 may specifically bind to cMet. For example, VH1 may specifically bind to CD3. For example, VH1 may specifically bind to CD19. For example, VH1 may specifically bind to HER2. For example, VH1 mayFor example, VH1 may specifically bind to CD28. For example, VH1 may specifically bind to CD38. For example, VH1 may specifically bind to Trop2. For example, VH1 may specifically bind to cMet. For example, VH2 may specifically bind to CD3. For example, VH2 may specifically bind to CD19. For example, VH2 may specifically bind to HER2. For example, VH2 may specifically bind to CD20. For example, VH2 may specifically bind to CD28. For example, VH2 may specifically bind to CD38. For example, VH2 may specifically bind to Trop2. For example, VH2 may specifically bind to cMet. For example, VH3 may specifically bind to CD3. For example, VH3 may specifically bind to CD19. For example, VH3 may specifically bind to HER2. For example, VH3 may specifically bind to CD20. For example, VH3 may specifically bind to CD28. For example, VH3 may specifically bind to CD38. For example, VH3 may specifically bind to Trop2. For example, VH4 may specifically bind to CD3. For example, VH4 may specifically bind to CD19. For example, VH4 may specifically bind to HER2. For example, VH4 may specifically bind to CD20. For example, VH4 may specifically bind to CD28. For example, VH4 may specifically bind to CD38. For example, VH4 may specifically bind to Trop2. For example, VH4 may specifically bind to cMet. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD28, VH2 and VL2 specifically bind to CD3, and VH3 and VL3 specifically bind to CD38. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD28, VH2 and VL2 specifically bind to CD38, and VH3 and VL3 specifically bind to CD3. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD3, VH2 and VL2 specifically bind to CD38, and VH3 and VL3 specifically bind to CD28.VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD3, VH2 and VL2 specifically bind to CD28, and VH3 and VL3 specifically bind to CD38. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD38, VH2 and VL2 specifically bind to CD28, and VH3 and VL3 specifically bind to CD3. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD38, VH2 and VL2 specifically bind to CD3, and VH3 and VL3 specifically bind to CD28. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD28, VH2 and VL2 specifically bind to CD19, and VH3 and VL3 specifically bind to CD38. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD28, VH2 and VL2 specifically bind to CD38, and VH3 and VL3 specifically bind to CD19. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD19, VH2 and VL2 specifically bind to CD38, and VH3 and VL3 specifically bind to CD28. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD19, VH2 and VL2 specifically bind to CD28, and VH3 and VL3 specifically bind to CD38. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD38, VH2 and VL2 specifically bind to CD28, and VH3 and VL3 specifically bind to CD19. In some embodiments, VH1 and VL1 of the antigen-binding polypeptide or antigen-binding polypeptide complex specifically bind to CD38, VH2 and VL2 specifically bind to CD19, and VH3 and VL3 specifically bind to CD28.The present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to EGFR and cMet. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to GP100 and CD3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to CD20 and CD3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to BCMA and CD3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to PDL1 and CTLA4. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to PD1 and LAG3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to PD1 and VEGF. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to DLL4 and VEGF. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to EGFR and HER3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to HER2. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to EpCAM and CD3. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to PDL1 and TGF-beta. For example, the present invention is directed to antigen-binding polypeptides or antigen-binding polypeptide complexes that specifically bind to PDL1 and TGF-beta. For example, the present invention is directed to an antigen-binding polypeptide or antigen-binding polypeptide complex that specifically binds to GPRC5D and CD3. For example, the present invention is directed to an antigen-binding polypeptide or antigen-binding polypeptide complex that specifically binds to CD123 and CD3. For example, the present invention is directed to an antigen-binding polypeptide or antigen-binding polypeptide complex that specifically binds to CD30 and CD16A. For example, the present invention is directed to an antigen-binding polypeptide or antigen-binding polypeptide complex that specifically binds to DLL3 and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of EGFR and cMet. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of GP100 and CD3.For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of CD20 and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of BCMA and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of PDL1 and CTLA4. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of PD1 and LAG3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of PD1 and VEGF. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of DLL4 and VEGF. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of EGFR and HER3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on HER2. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of EpCAM and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of PDL1 and TGF-beta. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of PDL1 and TGF-beta. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of GPRC5D and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of CD123 and CD3. For example, the antigen-binding polypeptide or antigen-binding polypeptide complex specifically binds to at least one epitope on each of CD30 and CD16A.For example, an antigen-binding polypeptide or an antigen-binding polypeptide complex may specifically bind to at least one epitope on each of DLL3 and CD3. For example, a polypeptide contained within an antigen-binding polypeptide or an antigen-binding complex may include VL1, VL2, VL3, VL4, VH1, VH2, VH3, ...

Claims

1. 1. An antigen-binding polypeptide complex comprising a first polypeptide and a second polypeptide, The first polypeptide is selected from the group consisting of: VL1-VL2-VH2-VH1-Fc, VH1-VH2-VL2-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc and having a structure represented by The second polypeptide is selected from the group consisting of: VL3-VL4-VH4-VH3-Fc, VH3-VH4-VL4-VL3-Fc, VL3-L5-VL4-L6-VH4-L7-VH3-Fc, VH3-L5-VH4-L6-VL4-L7-VL3-Fc, VL3-L5-VL4-L6-VH4-L7-VH3-L8-Fc, or VH3-L5-VH4-L6-VL4-L7-VL3-L8-Fc and having a structure represented by During the ceremony, VL1 is a first immunoglobulin light chain variable region; VL2 is a second immunoglobulin light chain variable region, VL3 is a third immunoglobulin light chain variable region, VL4 is a fourth immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region; VH2 is a second immunoglobulin heavy chain variable region; VH3 is a third immunoglobulin heavy chain variable region; VH4 is a fourth immunoglobulin heavy chain variable region; Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge; and L1, L2, L3, L4, L5, L6, L7, and L8 are amino acid linkers; The antigen-binding polypeptide complex.

2. VL1が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a first immunoglobulin light chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VL2が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a second immunoglobulin light chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VL3が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a third immunoglobulin light chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VL4が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a fourth immunoglobulin light chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VH1が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a first immunoglobulin heavy chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VH2が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a second immunoglobulin heavy chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VH3が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a third immunoglobulin heavy chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; VH4が、A2AR、APRIL、ATPDase、BAFF、BA FFR、BCMA、BlyS、BTK、BTLA、B7DC、B7H 1、B7H2、B7H3、B7H4、B7H5、B7H6、B7H7 、B7RP1、B7-4、C3、C5、CCL2、CCL3、CCL 4、CCL5、CCL7、CCL8、CCL11、CCL15、CC L17、CCL19、CCL20、CCL21、CCL25、CCR 3、CCR4、CD3、CD16A、CD19、CD20、CD24 、CD27、CD28、CD30、CD38、CD39、CD40、C D40L、CD47、CD52、CD70、CD80、CD86、C D123、CD133、CD137、CD137L、CD160、C D272、CEACAM5、CLEC9、CLEC91、CRTH2 、CSF-1、CSF-2、CSF-3、CXCL1、CXCL2、C XCL4、CXCL12、CXCL13、CXCR3、cMet、C TLA4、DLL3、DLL4、DNGR-1、E-カドヘリン、E GFR、ENTPD1、EpCAM、FCER1、FCER1A、F CER2、FGFR、FLAP、FOLH1、Gi24、GITR、G ITRL、GPR5、GP100、GPRC5D、HER2、HER 3、ICOSL、ICOS、HHLA2、HMGB1、HVEM、I DO、IFNa、IgE、IGF1R、IL2Rベータ、IL1、I LIA、IL1B、IL1F10、IL2、IL4、IL4Ra、I L5、IL5R、IL6、IL7、IL7Ra、IL8、IL9、I L9R、IL10、rhILlO、IL12、IL13、IL13R al、IL13Ra2、IL15、IL17、IL17Rb、IL18、IL22、IL23、IL25、I TGB4、ITK、KIR、LAG3、LAMP1、レプチン、LP FS2、MHCクラスII、MUC-1、MUC-16、NCR3L G1、NKG2D、NKp46、NTPDase-1、OX40、O X40L、PD-1、PD-L1、PD-L2、PROM1、S152 、SIRPアルファ、SISP1、SLC、SPG64、ST2、S TEAP1、STEAP2、Sykキナーゼ、TROP2、TACI 、㼴㼤㼯、㼴㼧㼦㼢㼥㼴㼡、㼴㼑㼔、㼴㼩㼧㼩㼴、㼴㼩㼭㼓、㼴㼬㼲、㼴㼬㼲㼒、㼴㼬㼲〔、㼴㼬㼲㼕、㼴㼬㼲㼙、㼴㼭㼥㼦㼑、㼴㼮㼦a、a fourth immunoglobulin heavy chain variable region that specifically binds to at least one epitope on at least one antigen selected from the group consisting of TNFRSF7, Tp55, TREM1, TSLP, TSLPR, TWEAK, VEGF, VISTA, Vstm3, and WUCAM; The antigen-binding polypeptide complex of claim 1.

3. The antigen-binding polypeptide complex of claim 1, wherein the immunoglobulin hinge comprises an upper hinge region, a middle hinge region, a lower hinge region, or a combination thereof.

4. 2. The antigen-binding polypeptide complex of claim 1, wherein the linkers L1, L2, L3, L4, L5, L6, L7, and / or L8 of the first polypeptide and / or the second polypeptide have a length of from about 1 amino acid to about 50 amino acids.

5. The linkers L1, L2, L3, L4, L5, L6, L7, and / or L8 are g, a, gss, asg, ggssg, gssgs, gtvaa, asggs, astgg, asggsg, ggsggssgss, sggsgssggs, ggsggsgsggggsasgsg, ggsggsgsggggsasgsg, gggssgggggsggsgs ggsgs, ggggsggsgsgsgggsasgsg, gggssggsgsggsgsgsggsgs, sggssggsgsggsgsggsgsssg, gsgssggg gsggsgsggsgsssg, ggggsgsggsgggssggggsggggsggggsggggsggggs, ggggsggggsggggsggggsgggg 2. The antigen-binding polypeptide complex of claim 1, comprising the amino acid sequence of sggggsggggsggggsggggs, ggggsgsggsgggssggggsggggsggggsggggsggggsggggsggggsggggsss, ggggsgsgggssggggsggggsggggsggggsggggsggggssss, ggsgg, gsggsagsgsggggsasgsg, ggggs, or gsggsggsgsggggsasgsg (SEQ ID NOS: 1-19 and 681-688), or a sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% identity to any one of SEQ ID NOS: 1-19 and 681-688.

6. The antigen-binding polypeptide complex of claim 1 , wherein the amino acid linker is non-immunogenic.

7. The antigen-binding polypeptide complex of claim 1 , wherein the amino acid linker does not contain a consensus T-cell epitope.

8. 2. The antigen-binding polypeptide complex of claim 1, wherein the Fc region comprises at least one knob-into-hole modification.

9. the antigen-binding polypeptide complex is an IgG1 or IgG4 antibody; and The knob-into-hole modification is, based on the EU numbering scheme, as follows: (i) Knob substitutions of S354C and T366W, and hole substitutions of Y349C, T366S, L368A, and Y407V; (ii) hole substitutions at L234A, L235A, and P239A; (iii) hole substitutions at L234A and L235A; (iv) M428L and N433S hole substitutions; (v) M252Y, S254T, and T256E hole substitutions, or (vi) combinations thereof Including, 9. The antigen-binding polypeptide complex of claim 8.

10. The antigen-binding polypeptide complex of claim 1, comprising a detectable label.

11. 11. The antigen-binding polypeptide complex of claim 10, wherein the detectable label is a radioactive label, a chemiluminescent label, a fluorescent label, an enzyme, or a peptide tag, or a combination thereof.

12. The antigen-binding polypeptide complex of claim 11, wherein the peptide tag is a polyhistidine tag consisting of about 4 to about 10 histidine residues.

13. The antigen-binding polypeptide complex of claim 12, wherein the polyhistidine tag consists of approximately 8 histidine residues.

14. 10. The antigen-binding polypeptide complex of claim 1, conjugated to a drug as an antibody-drug conjugate (ADC).

15. 15. The antigen-binding polypeptide complex of claim 14, wherein the agent is a cytotoxic agent, an immunomodulatory agent, an imaging agent, or a therapeutic protein, or a combination thereof.

16. Equilibrium dissociation constant (K Ｄ 2. The antigen-binding polypeptide complex of claim 1, which binds to an antigen at

17. An antibody or antigen-binding fragment thereof comprising the antigen-binding polypeptide complex of claim 1.

18. 18. The antibody or antigen-binding fragment thereof of claim 17, wherein the antibody is IgG, IgM, IgE, IgA, or IgD.

19. The antibody or antigen-binding fragment thereof of claim 18, wherein the IgG is IgG1, IgG2, IgG3, or IgG4.

20. The antigen-binding fragment may be Fab, scFab, Fab', F(ab') ２ 18. The antibody or antigen-binding fragment thereof of claim 17, which is an Fv, Fv, or scFv.

21. 18. The antibody or antigen-binding fragment thereof of claim 17, wherein the antibody is human or humanized.

22. A polypeptide having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 32-43, 62-79, 130-138, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, and 679, or having at least 90% identity, at least 95% identity, or 100% identity to the amino acid sequence of SEQ ID NO: 32 or 33 that does not contain the eight histidine residues at the C-terminus.

23. A polypeptide encoded by a polynucleotide having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 44-55, 80-97, 139-147, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, and 680.

24. A polynucleotide encoding the antigen-binding polypeptide complex of claim 1.

25. A polynucleotide having at least 90% identity, at least 95% identity, or 100% identity to any one of SEQ ID NOs: 44-55, 80-97, 139-147, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652, 654, 656, 658, 660, 662, 664, 666, 668, 670, 672, 674, 676, 678, and 680.

26. A polynucleotide encoding a polypeptide having at least 90%, at least 95%, or 100% identity to any one of SEQ ID NOs: 32-43, 62-79, 130-138, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, 655, 657, 659, 661, 663, 665, 667, 669, 671, 673, 675, 677, and 679, or encoding a polypeptide having at least 90%, at least 95%, or 100% identity to the amino acid sequence of SEQ ID NO: 32 or 33 without the eight histidine residues at the C-terminus.

27. A vector comprising the polynucleotide of claim 24.

28. A host cell comprising the polynucleotide of claim 24.

29. A chimeric antigen receptor (CAR) comprising the antigen-binding polypeptide complex of claim 1.

30. An immune cell comprising the CAR of claim 29.

31. (i) an antigen-binding polypeptide complex according to any one of claims 1 to 16; (ii) a pharmaceutically acceptable carrier; and 10. A pharmaceutical composition comprising:

32. A kit comprising an antigen-binding polypeptide complex according to any one of claims 1 to 16.

33. below: VL1-VL2-VH2-VH1-Fc-Fc, VH1-VH2-VL2-VL1-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-Fc, VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc-Fc, VL1-L1-VL2-L2-VH2-L3-VH1-L4-Fc-L5-Fc, or VH1-L1-VH2-L2-VL2-L3-VL1-L4-Fc-L5-Fc An antigen-binding polypeptide complex comprising a polypeptide having a structure represented by During the ceremony, VL1 is a first immunoglobulin light chain variable region; VL2 is a second immunoglobulin light chain variable region, VH1 is a first immunoglobulin heavy chain variable region; VH2 is a second immunoglobulin heavy chain variable region; Fc is a region comprising immunoglobulin heavy chain constant region 2 (CH2), immunoglobulin heavy chain constant region 3 (CH3), and optionally an immunoglobulin hinge; and L1, L2, L3, L4, and L5 are amino acid linkers; The antigen-binding polypeptide complex.

34. A pharmaceutical composition for treating or preventing a disease or condition in a subject in need thereof, comprising an antigen-binding polypeptide complex according to any one of claims 1 to 16 and 33.

35. A pharmaceutical composition for treating or preventing cancer in a subject, comprising an antigen-binding polypeptide complex according to any one of claims 1 to 16 and 33.

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