Agonist split antibody

Abstract

This application relates to an antigen-binding molecule pair, each molecule comprising a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain simultaneously binds to a target antigen, and the cytokine receptor-binding domain binds to a subunit of the cytokine receptor complex. Biparatopic assembly of the cytokine receptor-binding domain in the presence of a target antigen enables selective activation of the cytokine receptor and effectively mimics cytokine activity in a targeted manner.

Description

[Technical Field]

[0001] This application relates to an antigen-binding molecule pair, each molecule comprising a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain simultaneously binds to a target antigen, and the cytokine receptor-binding domain binds to a subunit of the cytokine receptor complex. Biparatopic assembly of the cytokine receptor-binding domain in the presence of a target antigen enables selective activation of the cytokine receptor and effectively mimics cytokine activity in a targeted manner. [Background technology]

[0002] In recent years, cancer immunotherapy has grown dramatically, and cancer immunotherapy is becoming a primary strategy for combating disease. For many cancer types, immune checkpoint modulators, including anti-PD-1, have become standard care. However, despite all the advances made in the field of cancer immunotherapy in recent years, a significant proportion of patients still do not respond to available immunotherapies due to intrinsic or adaptive mechanisms of resistance. Cancer immunotherapy patients with non-inflammatory immune symptoms are more likely to not respond to immunotherapy. Immune cell infiltration in tumors has been shown to correlate with a patient's ability to respond to immunotherapy treatments. The development of new therapies aimed at increasing immune cell infiltration and enhancing immunogenicity is essential for patients.

[0003] Alongside the developments mentioned above, cytokines are attracting significant interest as a potential cancer treatment. IFNγ (interferon-gamma or IFN-γ) responds to immune or inflammatory stimuli, and CD4 + and CD8 +IFNγ is a cytokine primarily produced by activated lymphocytes such as T cells, as well as natural killer (NK) cells. IFNγ is a homodimer, and its receptors (IFNγR1 and IFNγR2) are expressed across hematopoietic and non-hematopoietic cells. On the cell surface, IFNγR1 is stably expressed, while IFNγR2 is differentially expressed, regulating IFNγ. When IFNγ binds to its receptor, Janus kinases JAK1 and JAK2 are recruited and activated, phosphorylating and activating STAT1. Phosphorylated STAT1 translocates to the nucleus, binds to promoters, and regulates gene transcription under the control of IFNγ.

[0004] In contrast to other cancer therapies currently on the market or under development, IFNγ can act on both tumor cells and immune cells, including T cells and dendritic cells. The effects of IFNγ on various cell types offer several advantages, including: 1) enhanced expression of MHC-I molecules on the surface of both tumor cells and antigen-presenting cells; 2) recruitment of immune cells to tumor sites by inducing CXCL9, CXCL10, and CXCL11 production; and 3) increased tumor antigen cross-presentation, leading to enhanced anti-tumor immune responses. In addition to these effects, IFNγ promotes Th1 environment formation, monocyte differentiation, macrophage polarization, and angiogenesis. Because the IFNγ receptor is expressed across a wide range of cell types, the sink effect can reduce its activity.

[0005] IFNγ can also exhibit undesirable side effects. Furthermore, problems may arise regarding administration, bioavailability, and short half-life. Therefore, there is a need for novel molecules that can selectively activate the IFNγ pathway at tumor sites.

[0006] Targeting the interleukin-2 (IL-2) pathway as a cancer immunotherapy strategy has a long and branching history, serving as evidence of both the merits and complexities of this clinical approach. IL-2 is a cytokine that activates lymphocytes and natural killer (NK) cells. Clinical efficacy is often overshadowed by reports of IL-2-related toxicity associated with peripheral T cell activity and CD25-mediated complications.

[0007] Cytokines are potent immunomodulators that initiate signaling via receptor dimerization, but natural cytokines have several limitations as therapeutic agents. Low stability and difficulties in manufacturing processes are just some of them. It has been known for some time that antibodies can induce signaling on cells and thus replace natural ligands.

[0008] Recent literature provides growing evidence that combining heterodimeric cytokine receptors using both antibody- and non-antibody-based protein scaffolds is a viable strategy for mimicking the activity of native cytokines. For example, Moraga et al. provide an early example of using diabodies as surrogate ligands for the erythropoietin receptor (EpoR) (Moraga et al. Cell 160, 1196-1208 (2015)). Researchers at Teneobio combined heavy-chain-only antibodies (VHH fused to the Fc domain) against different epitopes on interleukin-2 receptors (IL-2Rβ and IL-2Rγ) with bispecific effectorless IgG4Fc (CH1 deletion) using knob-in-hole technology. Monospecific anti-IL-2Rβ or anti-IL-2Rγ alone or in combination were used with human CD8 +Although it did not induce STAT5 phosphorylation on T cells, the bispecific anti-IL-2Rβγ antibody showed varying levels of agonist activity (Harris, KE, et al. Sci Rep 11(1):10592 (2021)). A similar approach to that described by Teneobio was carried out by scientists at Synthekine, which described the functional induction of signaling between two interleukin receptors (IL-2Rβ and IL-2Rγ) via a single-domain antibody (sdAb; International Publication No. 2022 / 032040) and was reviewed by Saxton et al. (Saxton, RA, et al. Nat Rev Drug Discov. 22, 21-37 (2022)). This approach was further extended by researchers at Stanford University, where the authors presented a strategy for discovering cytokine surrogate agonists by using modular ligands such as VHH or scFv human for interleukin-2 / 15, type I interferon, and interleukin-10 receptors. Interestingly, they also identified functional non-native assemblies such as the IL-2Rβ / IL-10Rβ heterodimer (Yen, M., et al. Cell 185(8):1414-1430 e1419(2022)). The same authors also discuss a structure-based approach to decouple the pro-inflammatory and anti-inflammatory functions of interleukin-10 (Saxton, RA, et al. Science 371(6535)(2021)). Two academic research groups from the Czech Republic and Israel jointly reported the discovery of non-antibody-based scaffolds that mimic the cytokine IFNλ. Using combinatorial libraries derived from several established small protein scaffolds, variants capable of binding to IFNλR1 or IL-10Rβ and inducing functional signaling were identified (Kolarova, L., et al. FEBS J 289(9):2672-2684(2022)). The reduction in the size of the agonist module was addressed by researchers at Medikine.They obtained molecules selected from a peptide library through a screening designed to identify molecules that simultaneously bind to the Rα and γc subunits of the human IL-7 receptor. These peptides, with molecular weights less than 5 kDa and fused to the IgG1-Fc domain, exhibit similar biological properties to those of IL-7 in vitro and when administered to non-human primates (Dower, W., et al. Journal for Immuno Therapy of Cancer 8(Suppl 3):A341-A342(2020)).

[0009] Due to the multifaceted effects of cytokines, there is a need for novel approaches that selectively activate cytokine receptors and effectively mimic cytokine activity. [Overview of the project]

[0010] The present invention relates to a novel antigen-binding molecule comprising a cytokine receptor-binding domain that acts as a cytokine mimetic to selectively activate the receptor pathway under desired conditions of a biparatopic assembly on a target antigen.

[0011] The present invention relates to a pair of antigen-binding molecules that specifically bind to a target antigen, comprising: a) i) a first target-binding domain, ii) a first cytokine receptor-binding domain, and iii) an Fc domain; and b) i) a second target-binding domain, ii) a second cytokine receptor-binding domain, and iii) an Fc domain, wherein the first target-binding domain can bind to a first epitope on a target antigen, the second target-binding domain can bind to a second epitope on a target antigen, the first and second target-binding domains do not compete for binding to the target antigen, the first cytokine receptor-binding domain can bind to a first cytokine receptor subunit, and the second cytokine receptor-binding domain can bind to a second cytokine receptor subunit.

[0012] In one embodiment, the first and second target-binding domains are antibody fragments such as Fv, Fab, scFv, scFab molecules, or single-domain antibodies. In one embodiment, the first and second target-binding domains are Fab molecules. In one embodiment, the first target-binding domain includes a heavy chain variable domain (VH1), a light chain variable domain (VL1), a heavy chain constant domain (CH11), and a light chain constant domain (CL1), and the second target-binding domain includes a heavy chain variable domain (VH2), a light chain variable domain (VL2), a heavy chain constant domain (CH12), and a light chain constant domain (CL2). In one embodiment, the first and / or second target-binding domains are cross-Fab molecules.

[0013] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to tumor-associated antigens or T-cell antigens. In another embodiment, the first target-binding domain and the second target-binding domain specifically bind to FAP, PD-1, Her2, Her3, LAG-3, or EGFR. In one embodiment, a) the first target-binding domain includes VH1 of SEQ ID NO: 20 and VL1 of SEQ ID NO: 21, and the second target-binding domain includes VH2 of SEQ ID NO: 22 and VL2 of SEQ ID NO: 23, or b) the first target-binding domain includes VH1 of SEQ ID NO: 22 and VL1 of SEQ ID NO: 23, and the second target-binding domain includes VH2 of SEQ ID NO: 20 and VL2 of SEQ ID NO: 21, or c) the first target-binding domain includes VH1 of SEQ ID NO: 76 and SEQ ID NO: 77 d) The first target-binding domain includes VL1 of SEQ ID NO: 78 and VL2 of SEQ ID NO: 79, and the second target-binding domain includes VH2 of SEQ ID NO: 76 and VL2 of SEQ ID NO: 77, or e) The first target-binding domain includes VH1 of SEQ ID NO: 93 and VL1 of SEQ ID NO: 94, and the second target-binding domain includes VH2 of SEQ ID NO: 78 and VL2 of SEQ ID NO: 79 f) The first target-binding domain includes VH1 of SEQ ID NO: 78 and VL1 of SEQ ID NO: 79, and the second target-binding domain includes VH2 of SEQ ID NO: 93 and VL2 of SEQ ID NO: 94, or g) The first target-binding domain includes VH1 of SEQ ID NO: 130 and VL1 of SEQ ID NO: 131, and the second target-binding domain includes VH2 of SEQ ID NO: 132 and VL2 of SEQ ID NO: 133, or h) The first target-binding domain includes VH1 of SEQ ID NO: 132 i) The first target-binding domain includes H1 and VL1 of SEQ ID NO: 133, and the second target-binding domain includes VH2 of SEQ ID NO: 130 and VL2 of SEQ ID NO: 131, or j) The first target-binding domain includes VH1 of SEQ ID NO: 140 and VL1 of SEQ ID NO: 141, and the second target-binding domain includes VH2 of SEQ ID NO: 142 and VL2 of SEQ ID NO: 143,k) The first target-binding domain includes VH2 of SEQ ID NO: 140 and VL2 of SEQ ID NO: 141, and the second target-binding domain includes VH1 of SEQ ID NO: 114 and VL1 of SEQ ID NO: 115, and the second target-binding domain includes VH2 of SEQ ID NO: 116 and VL2 of SEQ ID NO: 117, or l) The first target-binding domain includes VH1 of SEQ ID NO: 116 and VL1 of SEQ ID NO: 117, and the second target-binding domain includes VH2 of SEQ ID NO: 114 and (m) The first target-binding domain includes VL2 of SEQ ID NO: 115 and VL1 of SEQ ID NO: 150, and the second target-binding domain includes VH2 of SEQ ID NO: 152 and VL2 of SEQ ID NO: 153, or (n) The first target-binding domain includes VH1 of SEQ ID NO: 152 and VL1 of SEQ ID NO: 153, and the second target-binding domain includes VH2 of SEQ ID NO: 150 and VL2 of SEQ ID NO: 151.

[0014] In one embodiment, both the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of an IFNγ receptor complex, an IL-2 receptor complex, or an IL-7 receptor complex. In one embodiment, a) the first cytokine receptor binding domain can bind to IFNγR1 and the second cytokine receptor binding domain can bind to IFNγR2, or b) the first cytokine receptor binding domain can bind to IFNγR2 and the second cytokine receptor binding domain can bind to IFNγR1, or c) the first cytokine receptor binding domain can bind to IL-2Rβ and the second cytokine receptor binding domain can bind to IL-2Rγ, or d) the first cytokine receptor binding domain can bind to IL-2Rγ and the second cytokine receptor binding domain can bind to IL-2Rβ, or e) the first cytokine receptor binding domain can bind to IL-2Rγ and the second cytokine receptor binding domain can bind to IL-7Rα, or f) the first cytokine receptor binding domain can bind to IL-7Rα and the second cytokine receptor binding domain can bind to IL-2Rγ.

[0015] In one embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain are antibody fragments such as Fv, Fab, scFv, scFab, or single-domain antibodies. In another embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain are VHH domains. In one embodiment, a) the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, or b) the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5, or c) the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, and SEQ ID NO: 60, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, or d) the first e) The cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 62, 63, 64 and 65, and the second cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 56, 57, 58 and 60, or f) The first cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191 and 192, and the second cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 61, 62, 63, 64 and 65, or f) The first cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 61, 62, 63, 64,The second cytokine receptor-binding domain contains an amino acid sequence selected from SEQ ID NO: 65, and SEQ ID NO: 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, and 192.

[0016] In one embodiment, a) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 3 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 7, or b) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 7 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 3, or c) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 60 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, or d) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 60, or e) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 186 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, or f) the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62 and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 186.

[0017] In one aspect, the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include a first Fc domain subunit and a second Fc domain subunit. In one aspect, the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are the Fc domains of IgG, particularly IgG1. In one aspect, the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are human Fc domains. In one aspect, the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include modifications that promote the association of the first subunit and the second subunit of the Fc domain. In one aspect, the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include one or more amino acid substitutions that reduce binding to Fc receptors and / or effector functions.

[0018] In one aspect, the cytokine receptor binding domains are fused to their respective fusion points via a peptide linker. In one aspect, the peptide linker includes an amino acid sequence selected from SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 279, SEQ ID NO: 280 or SEQ ID NO: 281.

[0019] In one embodiment, the first cytokine receptor binding domain is fused at its C-terminus to the N-terminus of VH1 or VL1 of the first target binding domain, and the second cytokine receptor binding domain is fused at its C-terminus to the N-terminus of VH2 or VL2 of the second target binding domain. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor binding domain, VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a second cytokine receptor binding domain, VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing VL2 and CL2 in order from the N-terminus to the C-terminus, or b) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and c) The first antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, a first cytokine receptor-binding domain, a third polypeptide containing VL1 and CL1, and the second antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12 and a second Fc domain subunit, and, in order from the N-terminus to the C-terminus, a third polypeptide containing a second cytokine receptor-binding domain, VL2 and CL2, or c) The first antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor-binding domain, VH1, CH11 and a second Fc domain subunit, and, in order from the N-terminus to the C-terminus, a third polypeptide containing VL1 and CL1, and the second antigen-binding molecule comprises, in order from the N-terminus to the C-terminus,The polypeptide comprises a second polypeptide containing CH12 and a second Fc domain subunit, and a third polypeptide containing a second cytokine receptor binding domain, VL2, and CL2, in order from the N-terminus to the C-terminus. VH1, VL1, CH11, and CL1 form the first target binding domain, and VH2, VL2, CH12, and CL2 form the second target binding domain.

[0020] In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor-binding domain, VH1, CH11 and a second Fc domain subunit, arranged sequentially from the N-terminus to the C-terminus, and a third polypeptide containing VL1 and CL1, arranged sequentially from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second cytokine receptor-binding domain, VH2, a) The first antigen-binding molecule comprises a first polypeptide containing CH12 and a second Fc domain subunit, and a third polypeptide containing VL2 and CL2 in order from the N-terminus to the C-terminus, or b) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a first cytokine receptor-binding domain, VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen The protobinding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus, or c) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and a first cytokine receptor binding domain, VH1, CH11 and The second antigen-binding molecule comprises a second polypeptide containing a second Fc domain subunit, and a third polypeptide containing VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a second cytokine receptor-binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus, or d) the first antigen-binding molecule isThe second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor-binding domain, VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing VH1 and CL1 in order from the N-terminus to the C-terminus, wherein the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and a second cytokine receptor-binding domain, VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus e) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a first cytokine receptor-binding domain, VH1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule contains a first Fc domain subunit The first antigen-binding molecule comprises a first polypeptide, a second polypeptide containing VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a second cytokine receptor-binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus, or f) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and a second polypeptide containing a first cytokine receptor-binding domain, VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12, and a second Fc domain subunit, and a third polypeptide containing a second cytokine receptor-binding domain, VL2, and CL2, in order from the N-terminus to the C-terminus, or g) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit,A second polypeptide comprising, in order from the N-terminus to the C-terminus, VL1, CH11 and a second Fc domain subunit, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, a first cytokine receptor binding domain, VH1 and CL1, wherein the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising, in order from the N-terminus to the C-terminus, a second cytokine receptor binding domain, VH2, CH12 and a second Fc domain subunit, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, VL2 and CL2, wherein VH1, VL1, CH11 and CL1 form a first target binding domain, and VH2, VL2, CH12 and CL2 form a second target binding domain.

[0021] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to FAP, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IFNγ receptor complex. Therefore, in one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 27, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 30, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 38, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or c) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, and sequence The second antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of number 32 and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 35, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 27, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33, or f) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 30, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31, or g) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28 and SEQ ID NO: 38 The second antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36, or h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 35.

[0022] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to EGFR, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IFNγ receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 260, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 256, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 122, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 255, or c) the first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 106 1) The antigen-binding molecule comprises polypeptide 1, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or d) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 257, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 254, or e) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 260,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 255, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 256, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106 The first antigen-binding molecule comprises a polypeptide, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 254, or (h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 257, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0023] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to PD-1, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 86, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 83, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 87; or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 92, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 91; or c) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28 and SEQ ID NO: 96 The second antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 95, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 99, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or f) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 100, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28 and the amino acid sequence of SEQ ID NO: 88 g) The first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 104, a second polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 99, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or h) the first The antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 95, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102, or i) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28 and the amino acid sequence of SEQ ID NO: 100 The second antigen-binding molecule comprises a second polypeptide and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or (j) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97, and the second antigen-binding molecule isIt comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 104.

[0024] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to LAG-3, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or c) the first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 106 1) The antigen-binding molecule comprises polypeptide 1, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or d) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157, or e) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106 The first antigen-binding molecule comprises a polypeptide, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or (h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157.

[0025] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to EGFR, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or c) the first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 106 1) The antigen-binding molecule comprises polypeptide 1, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or d) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157, or e) The antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106 The first antigen-binding molecule comprises a polypeptide, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or (h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157.

[0026] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to FAP, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109, or c) the first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 106 d) The first antigen-binding molecule comprises polypeptide 1, a second polypeptide containing the amino acid sequence of SEQ ID NO: 111, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or e) The first antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108, or e) The first antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110,The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 25, a second polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106 The first antigen-binding molecule comprises a polypeptide, a second polypeptide containing the amino acid sequence of SEQ ID NO: 111, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108, or h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0027] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to Her2, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 136, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 134, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 138, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 137, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 135, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 134, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 138, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 139.

[0028] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to Her3, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 146, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 144, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 148, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 147, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 145, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 144, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 148, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 149.

[0029] In one embodiment, the first target-binding domain and the second target-binding domain specifically bind to PD-1, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-7 receptor complex. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 194, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 193, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 195, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or c) the first antigen-binding molecule comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 106 The first antigen-binding molecule comprises polypeptide 1, a second polypeptide containing the amino acid sequence of SEQ ID NO: 196, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 197, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises polypeptide 1 containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 198,The second antigen-binding molecule comprises a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 199, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises SEQ ID NO: 28 The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 90, a second polypeptide containing the amino acid sequence of SEQ ID NO: 89, or f) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 200, a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and sequence number (a) The first antigen-binding molecule comprises a third polypeptide containing the amino acid sequence of SEQ ID NO. 89, or (b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO. 106, a second polypeptide containing the amino acid sequence of SEQ ID NO. 201, and a third polypeptide containing the amino acid sequence of SEQ ID NO. 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO. 28, a second polypeptide containing the amino acid sequence of SEQ ID NO. 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO. 89, or (h) the first antigen-binding molecule comprises SEQ ID NO. 10 The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of 6, a second polypeptide containing the amino acid sequence of SEQ ID NO: 202, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, wherein the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or i) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106 and a second polypeptide containing the amino acid sequence of SEQ ID NO: 203,The second antigen-binding molecule comprises a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or (j) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 204, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28 The peptide comprises a second polypeptide containing the amino acid sequence of SEQ ID NO: 90 and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or (k) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 205, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0030] In one embodiment, the first target-binding domain is fused to the N-terminus of the first Fc domain subunit at the C-terminus of CH11, the first cytokine receptor-binding domain is fused to the N-terminus of the second Fc domain subunit at its C-terminus, the second target-binding domain is fused to the N-terminus of the first Fc domain subunit at the C-terminus of CH12, and the second cytokine receptor-binding domain is fused to the N-terminus of the second Fc domain subunit at its C-terminus. In one embodiment, a) the first antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, a first polypeptide comprising VH1, CH11 and a first Fc domain subunit; a second polypeptide comprising VL1 and CL1 in order from the N-terminus to the C-terminus; and a third polypeptide comprising a first cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, a first polypeptide comprising VH2, CH12 and a first Fc domain subunit; a second polypeptide comprising VL2 and CL2 in order from the N-terminus to the C-terminus; and a third polypeptide comprising a second cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus; or b) the first antigen-binding molecule comprises, in order from the N-terminus to the C-terminus The first polypeptide comprises, in order from the N-terminus to the C-terminus, VL1, CH11, and a first Fc domain subunit; the second polypeptide comprises, in order from the N-terminus to the C-terminus, VH1, and CL1; and the third polypeptide comprises, in order from the N-terminus to the C-terminus, a first cytokine receptor binding domain and a second Fc domain subunit; the second antigen-binding molecule comprises, in order from the N-terminus to the C-terminus, VH2, CH12, and a first Fc domain subunit; the second polypeptide comprises, in order from the N-terminus to the C-terminus, VL2, and CL2; and the third polypeptide comprises, in order from the N-terminus to the C-terminus, a second cytokine receptor binding domain and a second Fc domain subunit; VH1, VL1, CH11, and CL1 form the first target-binding domain; and VH2, VL2, CH12, and CL2 are,A second target-binding domain is formed. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 32, a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 39, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 34, a second polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 34 c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 29, a second polypeptide containing the amino acid sequence of SEQ ID NO: 39, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40, or c) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and the amino acid sequence of SEQ ID NO: 25 The second antigen-binding molecule comprises a third polypeptide containing the amino acid sequence of SEQ ID NO: 238, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises the amino acid sequence of SEQ ID NO: 236 The first antigen-binding molecule comprises a first polypeptide containing the sequence, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or e) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 239, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 240 and a second polypeptide containing the amino acid sequence of SEQ ID NO: 34,or f) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 236, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and the amino acid sequence of SEQ ID NO: 25 The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or (h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 240, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 239, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0031] In one embodiment, a first target-binding domain is fused to the N-terminus of one of the Fc domain subunits at the C-terminus of CH11, a first cytokine receptor-binding domain is fused to the C-terminus of the same Fc domain subunit at its N-terminus, a second target-binding domain is fused to the N-terminus of one of the Fc domain subunits at the C-terminus of CH12, and a second cytokine receptor-binding domain is fused to the C-terminus of the same Fc domain subunit at its N-terminus. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11, a second Fc domain subunit and a first cytokine receptor binding domain in order from the N-terminus to the C-terminus, and a third polypeptide containing VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12, a second Fc domain subunit and a second cytokine receptor binding domain in order from the N-terminus to the C-terminus, and a third polypeptide containing VL2 and CL2 in order from the N-terminus to the C-terminus, or b) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit and in order from the N-terminus to the C-terminus The second antigen-binding molecule comprises, in order, a second polypeptide containing VL1, CH11, a second Fc domain subunit and a first cytokine receptor-binding domain, and, in order from the N-terminus to the C-terminus, a third polypeptide containing VH1 and CL1, and the second antigen-binding molecule comprises, the first polypeptide containing a first Fc domain subunit, and, in order from the N-terminus to the C-terminus, a second polypeptide containing VH2, CH12, a second Fc domain subunit and a second cytokine receptor-binding domain, and, in order from the N-terminus to the C-terminus, a third polypeptide containing VL2 and CL2, or c) the first antigen-binding molecule comprises, the first polypeptide containing a first Fc domain subunit, and, in order from the N-terminus to the C-terminus, a second polypeptide containing VL1, CH11, a second Fc domain subunit and a first cytokine receptor-binding domain,The second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12, a second Fc domain subunit and a second cytokine receptor-binding domain, and a third polypeptide containing VL2 and CL2, arranged in order from the N-terminus to the C-terminus. VH1, VL1, CH11, and CL1 form the first target-binding domain, and VH2, VL2, CH12, and CL2 form the second target-binding domain. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 41, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 45, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 43, or b) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 46, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 41, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 47, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 44 c) The first antigen-binding molecule comprises a polypeptide 2 and a third polypeptide containing the amino acid sequence of SEQ ID NO: 43, or the second antigen-binding molecule comprises a polypeptide 1 containing the amino acid sequence of SEQ ID NO: 241, a polypeptide 2 containing the amino acid sequence of SEQ ID NO: 246, and a polypeptide 3 containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a polypeptide 1 containing the amino acid sequence of SEQ ID NO: 241, a polypeptide 2 containing the amino acid sequence of SEQ ID NO: 249, and a polypeptide 3 containing the amino acid sequence of SEQ ID NO: 29, or the first antigen-binding molecule comprises a polypeptide 1 containing the amino acid sequence of SEQ ID NO: 241, a polypeptide 2 containing the amino acid sequence of SEQ ID NO: 242, and a polypeptide 3 containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule ise) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 245, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or e) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 250, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241 and a second polypeptide containing the amino acid sequence of SEQ ID NO: 253 The first antigen-binding molecule comprises a lipeptide and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or f) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 248, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 247, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) the first antigen-binding The molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 244, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 243, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or h) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241 and the amino acid sequence of SEQ ID NO: 252 The second antigen-binding molecule comprises a second polypeptide and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 251, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or i) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 42, and the second antigen-binding molecule isIt comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 245, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0032] In one embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain of the antigen-binding molecule pair are Fab molecules. In one embodiment, a) the first cytokine receptor binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 282 and a VL domain containing the amino acid sequence of SEQ ID NO: 283 or a VH domain containing the amino acid sequence of SEQ ID NO: 284 and a VL domain containing the amino acid sequence of SEQ ID NO: 285, and the second cytokine receptor domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 286 and a VL domain containing the amino acid sequence of SEQ ID NO: 287 or a VH domain containing the amino acid sequence of SEQ ID NO: 288 and a VL domain containing the amino acid sequence of SEQ ID NO: 289, or b) the first cytokine receptor binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 286 and a VL domain containing the amino acid sequence of SEQ ID NO: 287 or a VH domain containing the amino acid sequence of SEQ ID NO: 288 and a VL domain containing the amino acid sequence of SEQ ID NO: 289, and the second cytokine receptor domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 282 and a VL domain containing the amino acid sequence of SEQ ID NO: 283 or a VH domain containing the amino acid sequence of SEQ ID NO: 284 and a VL domain containing the amino acid sequence of SEQ ID NO: 285.16.

[0033] In one embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain of the antigen-binding molecule pair are scFv molecules. In one embodiment, a) the first cytokine receptor binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 290 and a VL domain containing the amino acid sequence of SEQ ID NO: 291, and the second cytokine receptor domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 292 and a VL domain containing the amino acid sequence of SEQ ID NO: 293, or b) the first cytokine receptor binding domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 292 and a VL domain containing the amino acid sequence of SEQ ID NO: 293, and the second cytokine receptor domain includes a VH domain containing the amino acid sequence of SEQ ID NO: 290 and a VL domain containing the amino acid sequence of SEQ ID NO: 291.

[0034] In one embodiment, the Fc domains of first and second antigen-binding molecules, which include a Fab or scFv molecule as the cytokine receptor-binding domain, include a first Fc domain subunit and a second Fc domain subunit. In one embodiment, the Fc domains of the first and second antigen-binding molecules are IgG, particularly IgG1 Fc domains. In one embodiment, the Fc domains of the first and second antigen-binding molecules are human Fc domains. In one embodiment, the Fc domains of the first and second antigen-binding molecules include modifications that facilitate association between the first and second subunits of the Fc domain. In one embodiment, the Fc domains of the first and second antigen-binding molecules include one or more amino acid substitutions that reduce binding to the Fc receptor and / or effector function. In one embodiment, the cytokine receptor-binding domains are fused to their respective fusion sites via a peptide linker. In one embodiment, the peptide linker includes the amino acid sequence of SEQ ID NO: 280 or SEQ ID NO: 281. In one embodiment, a) the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 267, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 266, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 272, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 271, or b) the The antigen-binding molecule of 1 comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 267, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 266, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 274, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 273, or c) the first antigen-binding molecule isThe first polypeptide comprises the amino acid sequence of SEQ ID NO: 269, the second polypeptide comprises the amino acid sequence of SEQ ID NO: 28, the third polypeptide comprises the amino acid sequence of SEQ ID NO: 265, and the fourth polypeptide comprises the amino acid sequence of SEQ ID NO: 268, and the second antigen-binding molecule comprises the first polypeptide comprising the amino acid sequence of SEQ ID NO: 268, the second polypeptide comprising the amino acid sequence of SEQ ID NO: 28, the third polypeptide comprising the amino acid sequence of SEQ ID NO: 270, and the fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 271, or d) First The antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 269, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 268. The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 274, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 273. In one embodiment, the first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 276, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 275. The second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 277, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 278. [Brief explanation of the drawing]

[0035] [Figure 1]Schematic diagram of antigens used for llama immunization, phage display, and screening for isolation of human IFNγR1, IFNγR2, IL-2Rβ, and IL-2Rγ-specific single-domain antibodies. For llama immunization, heterodimers (A) were generated from the extracellular domain (ECD) of human IFNγR1 fused to a biotinylated Fc knob and human IFNγR2 ECD fused to the Fc hole, and heterodimers (D) were generated from the ECD of human IL-2Rβ fused to a biotinylated Fc knob and human IL-2RγECD fused to the Fc hole. For phage display, monovalent human IFNγR1 (B), monovalent human IFNγR2 (C), monovalent human IL-2Rβ (E), monovalent human IL-2Rγ (F), and soluble Fc (G) were generated fused to biotinylated Fc. [Figure 2] A workflow for enriching single-domain antibodies with binding specificity to human cytokine receptor subunits using phage display. [Figure 3] Specificity screening of single-domain antibodies by ELISA. A random set of soluble VHH domains selected for human IFNγR1 specificity after 3 rounds of phage display was tested for binding to immobilized human IFNγR1-Fc and immobilized human Fc (A). VHH domains with human IFNγR2 specificity after 3 rounds of phage display were also tested for antigen specificity by ELISA (B). VHH domains selected for human IL-2Rβ specificity after 3 rounds of phage display were tested for binding to immobilized human IL-2Rβ-Fc and immobilized human Fc (C). VHH domains with human IL-2Rγ specificity after 3 rounds of phage display were also tested for antigen specificity by ELISA (D). The absorbance at 450 nm, showing the binding response to each target, is represented as a stacked bar graph. [Figure 4]Format conversion of single-domain antibody fragments. Round 3 VHH library variants with IFNγR1 or IL-2Rβ specificity were fused to the Fc-nob and cloned into mammalian cell expression vectors using the Gibson cloning method (A). Round 3 VHH library variants with IFNγR2 or IL-2Rγ specificity were fused to the Fc-hole using the same method (B). [Figure 5] A bispecific heavy chain antibody containing an IFNγR1-IFNγR2 or IL-2Rβ-IL-2Rγ VHH domain pair. The VHH domain with IFNγR1 or IL-2Rβ specificity was fused to an Fc knob, and the VHH domain targeting IFNγR2 or IL-2Rγ was fused to an Fc hole. The VHH moieties were fused to the Fc chain using a flexible 5 (G4S) linker. The bispecific heavy chain antibody was generated by knob-into-hole assembly of the Fc chain. The effector function of Fc was suppressed by incorporating the P329G LALA mutation into the CH2 domain. [Figure 6] Functional screening of bispecific heavy chain antibodies containing VHH domain pairs of IFNγR1-IFNγR2 or IL-2Rβ-IL-2Rγ. HEK-Blue IFNγ cells were incubated with IFNγ agonist bispecific heavy chain antibodies containing VHH domain pairs of IFNγR1 and IFNγR2, prepared using a 5×5 bispecific matrix (A). HEK-Blue IL-2 reporter cells were incubated with IL-2 agonist bispecific heavy chain antibodies containing VHH pairs of IL-2Rβ and IL-2Rγ, prepared using a 5×5 bispecific matrix (B). IFNγR activity and IL-2R activity were quantified by absorbance at 650 nm, and the multiple response / background for each treatment is shown. [Figure 7]Characterization of dose-dependent responses to IFNγ agonist heavy chain antibodies. Previously identified IFNγR agonists (Figure 6A, Table 1) were screened for dose-dependent IFNγR activity in HEK-Blue IFNγ cells characterized by absorbance at 650 nm. The responses of various IFNγ agonist heavy chain antibodies were grouped according to the anti-IFNγR1 VHH clone; i.e., in each graph, the anti-IFNγR2 VHH clone was variable, while the anti-IFNγR1 VHH clone remained constant: IFNγR1_1 clone (A), IFNγR1_2 clone (B), IFNγR1_3 clone (C), and IFNγR1_5 clone (D). Responses were compared with recombinant human IFNγ (black dashed line) and FAP-IFNγ (P1AF3574; light gray dashed line). [Figure 8] Comparison of EC50 values ​​of IFNγ agonist heavy chain antibodies. EC50 values ​​were obtained from the HEK-Blue assay using GraphPad Prism software (Figure 7). Values ​​are shown in nanomolar concentrations, and recombinant human IFNγ is highlighted as a reference (dotted line). [Figure 9] Characterization of dose-dependent responses to IL-2R agonist heavy chain antibodies. Previously identified IL-2R agonists (Figure 6B, Table 2) were screened for dose-dependent IL-2R agonism in HEK-Blue IL-2 cells. Anti-PD1-IL2v (P1AE4422) was used as a reference molecule, and each molecule was incubated with reporter cells at the following concentrations: 20 nM, 0.8 nM, and 0.032 nM from left to right. IL-2R activity was quantified by absorbance at 650 nm; the mean value from technical triplets is shown with error bars representing the standard deviation. [Figure 10]MHC-I and PD-L1 expression on tumor cells after 72-hour treatment with IFNγ agonists. Previously identified IFNγR agonists (P1AH1877-P1AH1890) and reference molecules (recombinant human IFNγ and FAP-IFNγ) were incubated with tumor cells at the following treatment concentrations: 100, 10, 1, and 0.1 nM for each treatment (left to right). Measurements were blanked out and normalized to the recombinant human IFNγ response at the highest concentration (100 nM). Responses for MHC-I expression on MKN45 cells (A), PD-L1 expression on MKN45 cells (B), MHC-I expression on Bxpc3 cells (C), PD-L1 expression on Bxpc3 cells (D), MHC-I expression on CorL105 cells (E), and PD-L1 expression on CorL105 cells (F) are shown. [Figure 11] Concept of a split dual-targeting IFNγR agonist. FAP-dependent biparatopic assembly of a split IFNγ mimetic (A). Note that for simplicity of explanation, only one monomer of the FAP dimer is shown in the inset. The VHH domain, which has IFNγR1 specificity and IFNγR2 specificity, was fused to the N-terminus of two different anti-FAP binding domains via a VH fusion site or a VL fusion site, resulting in eight conformations in the substitution space: IFNγR1-VHH fused to the VH of FAP binder 1 (B), IFNγR2-VHH fused to the VH of FAP binder 2 (C), and fused to the VL of FAP binder 1. IFNγR1-VHH(D), IFNγR2-VHH(E) fused to the VL of FAP binder 2, IFNγR2-VHH(F) fused to the VH of FAP binder 1, IFNγR1-VHH(G) fused to the VH of FAP binder 2, IFNγR2-VHH(H) fused to the VL of FAP binder 1, and IFNγR1-VHH(I) fused to the VL of FAP binder 2. All molecules have the same Fc properties as shown in Figure 5. [Figure 12]Evaluation of IFNγR activity mediated by FAP-dependent split IFNγR agonists. Eight split IFNγ mimes (Figure 11B-I) were paired in biparatopic assembly combinations and tested for IFNγR agonism characterized by MHC-I and PD-L1 upregulation. FAP-negative A549 cells were tested for untargeted, nonspecific MHC-I (A, B) and PD-L1 upregulation (C, D) by split IFNγ mimes. A549 FAP-positive cells were co-cultured with differentially labeled A549 FAP-negative cells to evaluate FAP-dependent activity of IFNγ mimes in cis and trans states: MHC-I upregulation in cis (E and F), PD-L1 upregulation in cis (G and H), MHC-I upregulation in trans (I and J), and PD-L1 upregulation in trans (K and L). Recombinant IFNγ and the IFNγ agonist heavy chain antibody P1AH1884 (shown in Figure 5) were used as references. Median fluorescence intensity (MFI) of MHC-I and PD-L1 expression levels was analyzed using FlowJo. The mean values ​​from technical replication are shown with error bars representing the standard deviation. [Figure 13]Further formats of split dual-targeting IFNγR agonists. Four conformations were obtained by detaching the VHH domain, which has IFNγR1 specificity and IFNγR2 specificity, from the anti-FAP binder via fusion to the N-terminus of the opposite Fc chain: IFNγR1 and FAP binder 1 (A), IFNγR2 and FAP binder 2 (B), IFNγR2 and FAP binder 1 (C), and IFNγR1 and FAP binder 2 (D). Alternatively, four conformations were obtained by detaching the VHH domain, which has IFNγR1 specificity and IFNγR2 specificity, from the anti-FAP binder via fusion to the C-terminus of the same Fc chain: IFNγR1 and FAP binder 1 (E), IFNγR2 and FAP binder 2 (F), IFNγR2 and FAP binder 1 (G), and IFNγR1 and FAP binder 2 (H). The anti-FAP binders in Figure 13E-H include the CrossFab VH / VL operation (binder 1) and the CH1-CL charge (binder 2), which then allow for further pairing. All molecules have the same Fc properties as those described in Figure 5. [Figure 14]Evaluation of IFNγR activity mediated by further FAP-dependent split IFNγR agonists. Four combinations of molecules and reference molecules shown in Figure 13 were tested for IFNγR agonism characterized by MHC-I and PD-L1 upregulation. A549 cells lacking FAP expression were tested for FAP-independent MHC-I upregulation (A) and PD-L1 upregulation (B) of split IFNγ mimes. A549 FAP-positive cells were co-cultured with differentially labeled A549 FAP-negative cells, and the FAP-dependent activity of IFNγ mimes was evaluated by cis-MHC-I upregulation (C), cis-PD-L1 upregulation (D), trans-MHC-I upregulation (E), and trans-PD-L1 upregulation (F). Recombinant IFNγ and IFNγ agonist heavy chain antibody (P1AH1884) were used as reference molecules. Median fluorescence intensity (MFI) of MHC-I and PD-L1 expression levels was analyzed using FlowJo. Mean values ​​from technical replication are shown with error bars representing the standard deviation. [Figure 15]Concept of split FAP-targeted IFNγ agonists with different linker lengths. A single VHH domain with specificity for either IFNγR1 or IFNγR2 was manipulated to ligate to the hinge region at the N-terminus (top) or C-terminus (bottom) of the Fc region within an antibody construct containing an FAP-binding Fab domain. The construct utilized one of two different FAP-binding Fab domains that did not compete for binding. To ligate the Fc region to the VHH domain, a flexible linker consisting of 5, 15, or 25 amino acids, indicated by a number following a semicolon after the molecular ID, was used. This resulted in a set of 24 unique conformations that can be paired to produce 12 functional FAP-targeted IFNγ agonists: P1AJ8564+P1AJ8567, P1AJ8560+P1AJ8563 and P1AJ8568+P1AJ8571(A); P1AJ8566+P1AJ8565, P1AJ85 62+P1AJ8561 and P1AJ8570+P1AJ8569(B); P1AJ8576+P1AJ8579, P1AI0187+P1AJ8575 and P1AJ8580+P1AJ8583(C); P1AJ8578+P1AJ8577, P1AJ8574+P1AJ8573 and P1AJ8582+P1AJ8581(C). [Figure 16]Evaluation of IFNγR activity mediated by further FAP-dependent split IFNγR agonists. Twelve combinations of molecules and reference molecules shown in Figure 15 were tested for IFNγR agonism characterized by MHC-I upregulation. The test compounds were incubated alone with A549 FAP-negative cells (untargeted conditions; A and D) or in combination with A549 FAP-positive cells, where cis-signaling (FAP binding to A549 FAP-positive cells and activity to A549 FAP-positive cells; B and E) and trans-signaling (FAP binding to A549 FAP-positive cells and activity to A549 FAP-negative cells; C and F) were evaluated. Recombinant IFNγ and IFNγ agonist heavy chain antibody (P1AH1884) were used as reference molecules. Median fluorescence intensity (MFI) of MHC-I expression levels was analyzed by FlowJo. Mean values ​​from technical replication are shown with error bars representing the standard deviation. [Figure 17] The concept of a split EGFR-targeted IFNγ agonist. VHH domains having IFNγR1 specificity and IFNγR2 specificity can be fused to the N-terminus of two different anti-EGFR binding domains via a VH fusion site or a VL fusion site to obtain eight distinct format configurations (P1AK 5514-P1AK5521), which can then be paired to produce the eight functional assemblies shown: P1AK5520+P1AK5519(A), P1AK5516+P1AK5515(B), P1AK5521+P1AK5518(C), P1AK5517+P1AK5514(D), P1AK5520+P1AK5515(E), P1AK5516+P1AK5519(F), P1AK5521+P1AK5514(G), and P1AK5517+P1AK5518(H). [Figure 18] Functional activity of EGFR-targeted IFNγ agonists. Test compounds were incubated with PBMCs and target cells, and CXCL10 production was evaluated after 24 hours of stimulation. Recombinant IFNγ was used as a reference. Mean + / -SD of technical replication is shown for each molecular concentration tested. [Figure 19]Concept of a split dual-targeting IL-2 agonist. PD-1-dependent biparatopic assembly of a split IL-2 mimetic (A). VHH domains with IL-2Rβ specificity and IL-2Rγ specificity were fused to the N-terminus of two different anti-PD-1 binding domains via VH or VL fusion sites: IL-2Rβ-VHH fused to VH of PD-1 binder 1 (B), IL-2Rγ-VHH fused to VL of PD-1 binder 2 (C), IL-2Rβ-VHH fused to VL of PD-1 binder 1 (D), and IL-2Rγ-VHH fused to VH of PD-1 binder 2 (E). IL-2 agonist heavy chain antibody (F) and PD-1-IL2v (G) were used as reference molecules. All molecules have the same Fc properties as shown in Figure 5. [Figure 20] Phosphorylation of STAT5 on CD4 T cells after 15-minute and 60-minute incubation with an IL-2R agonist. Activated T cells expressing PD-1 (PD-1+ subset) and activated T cells pre-blocked with anti-PD-1 antibody (PD-1- subset) were differentially labeled and treated with two combinations of split-bitargeted IL-2 mimetic: P1AH6850+P1AH6813(AD) and P1AH6814+P1AI1593(EH). IL-2 mimic heavy chain antibody (P1AH1177) and PD1-IL2v (P1AE4422) were used as reference molecules. MFI and frequency of STAT5-P+ cells were measured by FACS, showing responses for the PD-1+ subset (solid line) and PD-1- subset (dashed line). [Figure 21] Flow cytometry gating strategies and baseline receptor expression. PBMCs isolated from two healthy donors were stained with survival dye and fluorescent dye conjugated antibodies against cell surface antigens. Gating strategies for identifying five distinct cell types: NK cells and NKT cells (A); CD4 T cells and CD8 T cells (B); and γδ T cells (C). Representative plots showing co-expression of PD-1 and IL-2Rβ on CD8 T cells (D), γδ T cells (E), and NK cells (F). [Figure 22]CD25 expression on PBMC subsets induced by PD-1-targeted IL-2R agonists. The frequencies of CD8 T cells (A), γδ T cells (B), and NK cells (C) expressing CD25 were determined by flow cytometry after 5 days of co-incubation of the test compound with newly isolated CTV-labeled PBMCs. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. Mean + / - SEM values ​​of two combined PBMC donors and technical replications are shown for each molecular concentration tested. [Figure 23] Proliferation of PBMC subsets induced by PD-1-targeted IL-2R agonists. The frequencies of proliferating CD8 T cells (A), γδ T cells (B), and NK cells (C) were determined by flow cytometry after 5 days of co-incubation of the test compound with newly isolated CTV-labeled PBMCs. Proliferating cells were identified based on cell division visualized by CTV staining. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. Mean + / - SEM values ​​of two combined PBMC donors and technical replications are shown for each molecular concentration tested. [Figure 24]This report presents the results of efficacy studies evaluating PD-1-targeted IL-2R agonists (P1AI1593 and P1AH6814), PD1-IL2v immunoconjugate (P1AE4422), and pembrolizumab in combination with FOLR1-TCB Mab. Tumor growth inhibition in an orthotopic xenograft model of the mammary gland was studied by injecting BC004 human breast carcinoma PDX cells into the mammary fat pads of humanized BRGS47 mice. The antibody doses (mg / kg) injected per mouse were as follows: P1AI1593, P1AH6814, and pembrolizumab 1 mg / kg; PD1-IL2v 0.1 mg / kg; and FOLR1-TCB 0.3 mg / kg. Antibodies were administered intravenously once weekly for four weeks. The combination of FOLR1-TCB 0.3 mg / kg + PD1-IL2-targeted IL-2R agonist (P1AI1593 (day 1) + P1AH6814 (day 2)) mediated superior efficacy in inhibiting tumor growth compared to FOLR1-TCB monotherapy and other combination groups: FOLR1-TCB 0.3 mg / kg + PD1-IL2v and FOLR1-TCB 0.3 mg / kg + pembrolizumab. [Figure 25] Concept of a split PD-1 targeted IL-2R agonist using an alternative anti-PD-1 Fab binder. By fusing VHH domains with IL-2Rβ specificity and IL-2Rγ specificity to the N-terminus of two different anti-PD-1 binding domains via VH or VL fusion sites, eight distinct format configurations (P1AK2599; P1AK2798; P1AK2799; P1AK2802; P1AK2803; P1AK2806; P1AK2809; P1AK2810) can be obtained, and these can be paired to produce the eight functional assemblies shown: P1AK2802+P1AK2599(A), P1AK2809+P1AK2799(B), P1AK2803+P1AK2806(C), P1AK2810+P1AK2798(D), P1AK2809+P1AK2599(E), P1AK2802 P1AK2 799(F), P1AK2810+P1AK2806(G), and P1AK2803+P1AK2798(H). [Figure 26]Functional activity of PD-1-targeted IL-2R agonists using alternative anti-PD-1 Fab binders. Test compounds were incubated with HEK Blue IL-2 wt cells (A) or HEK Blue IL-2 human PD-1 cells (B) at 37°C and 5% CO2 for 21 hours. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by absorbance at 650 nm using QUANTI-Blue reagent. Mean absorbance values ​​+ / - SEM of technical replications are shown for each molecular concentration tested. [Figure 27] Concept of a split LAG-3 targeted IL-2R agonist. By fusing VHH domains with IL-2Rβ specificity and IL-2Rγ specificity to the N-terminus of two different anti-LAG-3 binding domains via a VH fusion site or a VL fusion site, eight distinct format configurations (P1AJ5654-P1AJ5661) can be obtained, and these can be paired to produce the eight functional assemblies shown: P1 AJ5654+P1AJ5661(A), P1AJ5658+P1AJ5657(B), P1AJ5655+P1AJ5660(C), P1AJ5659+P1AJ5656( D), P1AJ5658+P1AJ5661(E), P1AJ5654+P1AJ5657(F), P1AJ5659(G) and P1AJ5655+P1AJ5656(H). [Figure 28] Functional activity of LAG-3-targeted IL-2R agonists. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. The frequency of STAT-5 phosphorylation (A) and MFI (B) were determined by flow cytometry after 60-minute co-incubation of the test compound with CD4 T cells pre-activated for 3 days with plate-bound anti-CD3 antibody and soluble anti-CD28 antibody. [Figure 29]The concept of a split FAP-targeted IL-2R agonist. By fusing VHH domains with IL-2Rβ specificity and IL-2Rγ specificity to the N-terminus of two different anti-FAP binding domains via a VH fusion site or VL fusion site, eight distinct format configurations (P1AJ5092-P1AJ5099) can be obtained, and these can be paired to produce eight functional assemblies: P1AJ5096+P 1AJ5099(A), P1AJ5092+P1AJ5095(B), P1AJ5097+P1AJ5098(C), P1AJ5093+P1AJ5094(D), P1AJ50 96+P1AJ5095(E), P1AJ5092+P1AJ5099(F), P1AJ5097+P1AJ5094(G) and P1AJ5093+P1AJ5098(H). [Figure 30] Functional activity of FAP-targeted IL-2R agonists. Test compounds were incubated with A549 FAP-negative target cells (A) or A549 FAP-positive target cells (B) and IL-2Rβγ bioassay cells at 37°C and 5% CO2 for 20 hours. PD1-IL2v (P1AE4422), FAP-IL2v (P1AA5355), and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by luminescence using Bio-Glo NL reagent and normalized to background signal. The mean multiplier change in technical replication RLU values ​​+ / - SEM are shown for each molecular concentration tested. [Figure 31]The concept of a split EGFR-targeted IL-2R agonist. By fusing VHH domains with IL-2Rβ specificity and IL-2Rγ specificity to the N-terminus of two different anti-EGFR binding domains via a VH fusion site or VL fusion site, eight distinct format configurations (P1AJ4165-P1AJ4172) can be obtained, and these can be paired to produce eight functional assemblies: P1AJ4169+ P1AJ4172(A), P1AJ4165+P1AJ4168(B), P1AJ4170+P1AJ4171(C), P1AJ4166+P1AJ4167(D), P1AJ4169+P1AJ4168(E), P1AJ4165+P1AJ4172(F), P1AJ4170+P1AJ4167(G), and P1AJ4166+P1AJ4171(H). [Figure 32] Functional activity of EGFR-targeted IL-2R agonists. EGFR expression on target cells by flow cytometry (A). Test compounds were incubated with A431 target cells (B), OE19 target cells (C), or LoVo target cells (D) and IL-2Rβγ bioassay cells at 37°C and 5% CO2 for 20 hours. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by luminescence using Bio-Glo NL reagent and normalized to background signal. Mean multiplier change RLU values ​​+ / - SEM are shown for each molecular concentration tested. [Figure 33]Concept of split HER2-targeted and HER3-targeted IL-2R agonists. VHH domains with IL-2Rβ-specificity and IL-2Rγ-specificity were fused to the N-terminuses of two different anti-HER2 binding domains and two different anti-HER3 binding domains. The VH or VL fusion site was selected for each Fab based on structural data. Four format configurations (P1AJ 4173-P1AJ4176) and two paired assemblies were designed for HER2: P1AJ4174+P1AJ4175(A) and P1AJ4173+P1AJ4176(B). Four format configurations (P1AJ4177-P1AJ4180) and two paired assemblies were selected for HER3: P1AJ4178+P1AJ4179(C) and P1AJ4177+P1AJ4180(D). [Figure 34] Functional activity of HER2-targeted and HER3-targeted IL-2R agonists. Test compounds were incubated with MDA-MB-231-targeted cells (A) or OE19-targeted cells (B) and IL-2Rβγ bioassay cells at 37°C and 5% CO2 for 20 hours. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by luminescence using Bio-Glo NL reagent and normalized to background signal. For each molecular concentration tested, the mean multiplier change in technical replication RLU values ​​+ / - SEM are shown. [Figure 35]Concept of a split PD-1 targeted IL-2R agonist using alternative anti-PD-1 single-domain binders. VHH domains with IL-2Rβ specificity and IL-2Rγ specificity can be fused to the N-terminus of four different anti-PD-1 single-domain binders (P1AI0063; P1AK3046; P1AK3048; P1AK3051-P1AK3054), and these can be paired to produce the six functional assemblies shown: P1AK3052+P1AK3048(A), P1AI0063+P1AK3051(B), P1AK6054+P1AK3048(C), P1AI0063+P1AK3053(D), P1AK3054+P1AK3046(E), and P1AK3052+P1AK3046(F). [Figure 36] Functional activity of PD-1-targeted IL-2R agonists using alternative anti-PD-1 single-domain binders. Test compounds were incubated with HEK Blue IL-2 wt cells (A) or HEK Blue IL-2 human PD-1 cells (B) at 37°C and 5% CO2 for 20 hours. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by absorbance at 650 nm using QUANTI-Blue reagent. Mean absorbance values ​​+ / - SEM of technical replications are shown for each molecular concentration tested. [Figure 37]Concept of a split PD-1 targeted IL-2R agonist using an anti-IL2R Fab binder or an anti-IL2R scFv binder. Fab fragments having IL-2Rβ specificity and IL-2Rγ specificity were fused to the N-terminus of two anti-PD1 binding domains with different paratopes via a VH fusion site or a VL fusion site. P1AL9238(A) and P1AL9239(B) each contain PD1 binder 2 (FV003451 paratope; 1040 binder) and two different IL2Rγ binders FV018863 and FV018864, respectively, while P1AL9240(C) and P1AL9241(D) each contain PD1 binder 1 (FV000363 paratope; 0376 binder) and two different IL2Rβ binders FV002203 and FV018866, respectively. These can be paired to produce the following four functional assemblies: P1AL9238+P1AL9240, P1AL9238+P1AL9241, P1AL9239+P1AL9240, and P1AL9239+P1AL9241. Alternatively, scFv fragments having IL-2Rβ specificity and IL-2Rγ specificity were fused to the N-terminus of two anti-PD1 binding domains having different paratopes via a VH or VL fusion site. P1AL9310(E) contains PD1 binder 2 (FV003451 paratope) and IL2Rγ binder FV018865, while P1AL9311(F) contains PD1 binder 1 (FV000363 paratope) and IL2Rβ binder FV018866. These can be paired to produce the following functional assembly: P1AL9310+P1AL9311. [Figure 38]Functional activity of PD-1-targeted IL-2R agonists using anti-IL2R Fab and scFv binders. Test compounds were incubated with HEK Blue IL-2 wt cells (A) or HEK Blue IL-2 human PD-1 cells (B) at 37°C and 5% CO2 for 18 hours. An IL-2 agonist heavy chain antibody (Fc-VHH; P1AH1177) was used as a reference. An unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by absorbance at 650 nm using QUANTI-Blue reagent. The multiplicative change in IL-2R signaling against background + / - SEM of technical replication for each molecular concentration tested is shown. [Figure 39] Schematic diagram of antigens and workflow for isolating IL-7Rα-specific single-domain antibodies. The extracellular domain (ECD) of human IL-7Rα fused to a biotinylated Fc knob-into-hole (P1AI1009, represented by A) was generated for alpaca immunization, phage display, and screening. For phage display, a soluble human Fc knob-into-hole (P1AD4290, represented by B) was generated. Workflow (C) for enrichment of single-domain antibodies with binding specificity to IL-7Rα by phage display. [Figure 40] Specificity screening of single-domain antibodies by ELISA. Soluble VHH domains enriched with IL-7Rα specificity were randomly selected and screened for binding to immobilized human IL-7Rα-Fc and immobilized human Fc. After 3 rounds of phage display, VHH from Library 1 (A) and Library 2 (B) were tested for antigen specificity. After 4 rounds, enriched VHH from Library 1 (C) and Library 2 (D) were also tested for specificity by ELISA. Absorbance at 450 nm, indicating the binding response to each target, is represented as a stacked bar graph. [Figure 41]Format conversion of single-domain antibody fragments to bispecific heavy chain antibodies. Round 3 and Round 4 IL-7Rα-specific VHHs from libraries 1 and 2 were fused to an Fc knob and cloned into a mammalian cell expression vector using the Gibson cloning method (A). Previously identified VHHs with IL-2Rγ specificity were fused to an Fc hole using the same method (B). Bispecific heavy chain antibodies containing the IL-7Rα-IL-2RγVHH domain pair were generated by knob-into-hole assembly of a flexible 5 (G4S) linker for fusion of the Fc chain and the VHH portion to the Fc chain. The effector function of Fc was suppressed by incorporating the P329G LALA mutation into the CH2 domain (C). [Figure 42] Functional characterization of dose-dependent responses to IL-7R agonists. Bispecific heavy-chain antibodies containing IL-7Rα specificity and IL-2Rγ specificity were screened for dose-dependent IL-7R agonism in HEK Blue IL-7 cells. Recombinant IL-7 was used as a reference, and an unspecific IgG (P1AA5879) was used as a negative control. Each molecule was incubated with reporter cells at the following concentrations: 20 nM, 2 nM, 0.2 nM, and 0.02 nM from left to right. IL-7R activity was quantified by absorbance at 650 nm, showing the multiple response / background for each treatment. Graphs were separated according to four different IL-2Rγ-specific VHH clones: IL2Rγ_2(A), IL2Rγ_3(B), IL2Rγ_4(C), and IL2Rγ_5(D). [Figure 43]Concept of a split PD-1 targeted IL-7R agonist. PD-1-dependent biparatopic assembly of a split IL-7 mimetic (A). VHH domains with IL-7Rα specificity and IL-2Rγ specificity were fused to the N-terminus of two different anti-PD-1 binding domains via VH or VL fusion sites: IL-7Rα-VHH fused to the VL of PD-1 binder 1 (B), and IL-2Rγ-VHH fused to the VH of PD-1 binder 2 (C). The IL-7Rα molecules contained AAA mutations (I253A; H310A; H435A, numbering according to the Kabat EU index) in the CH2 and CH3 domains of the Fc hole to facilitate protein A purification. The effector function of Fc was suppressed in all molecules by incorporating the P329G LALA mutation into the CH2 domain. [Figure 44] Functional activity of PD-1-targeted IL-7R agonists. PD1-IL2v (P1AE4422) and Fc-VHH (P1AH1177) were used as references. Unbound DP47 antibody (P1AD3966) was used as a negative control. For blood donors 1 (A and B) and 2 (C and D), the frequency of STAT-5 phosphorylation was determined by flow cytometry after 60-minute co-incubation of the test compound with CD4 T cells pre-activated for 3 days with plate-bound anti-CD3 antibody and soluble anti-CD28 antibody. [Figure 45]The concept of a split PD-1-targeted IL-2R agonist having a monoparatopic assembly on PD-1. A VHH domain with IL-2Rβ specificity and IL-2Rγ specificity is fused to the N-terminus of two different anti-PD-1 binding domains via a VH fusion site or a VL fusion site, thereby achieving the same anti-PD-1 Eight individual format configurations of IL-2R agonist pairs containing a Fab binder (monoparatopic assembly) can be obtained and paired to produce eight monoparatopic assemblies: P1AK2802+P1AK2810(A), P1AK2809+P1AK2803(B), P1AK2802+P1AK2803(C), P1AK2809+P1AK2810(D), P1AK2798+P1AK2599(E), P1AK2806+P1AK2799(F), P1AK2798+P1AK2799(G), and P1AK2806+P1AK2599(G). [Figure 46] Functional activity of monoparatopic versus biparatopic PD-1-targeted IL-2R agonists. Test compounds were incubated with HEK Blue IL-2 wt (A) or HEK Blue IL-2 human PD-1 cells (B) at 37°C and 5% CO2 for 21 hours. Two biparatopic PD-1-targeted IL-2R agonists (P1AH6814+P1AI1593 and P1AH6814+P1AK2802) and an IL-2 mimic heavy chain antibody (Fc-VHH; P1AH1177) were used as references. An unbound DP47 antibody (P1AD3966) was used as a negative control. IL-2R signaling was measured by absorbance at 650 nm using QUANTI-Blue reagent. Mean absorbance values ​​+ / - SEM of technical replicates are shown for each molecular concentration tested. [Modes for carrying out the invention]

[0036] definition Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have meanings generally understood by those skilled in the art. Furthermore, unless otherwise specifically required by context, singular terms shall include plural forms and plural terms shall include singular forms. The methods and techniques of this disclosure are generally carried out in accordance with conventional methods well known in the art. Generally, the nomenclature and techniques used in connection with biochemistry, enzymology, molecular biology, and cell biology, microbiology, genetics, and protein and nucleic acid chemistry, as well as hybridization, as described herein are well known and commonly used in the art.

[0037] The terms "a," "an," and "the" generally refer to multiple objects unless the context explicitly indicates otherwise.

[0038] As used herein, the terms “first,” “second,” “third,” or “fourth” with respect to binding molecules, epitopes, polypeptides, etc., are used for convenience to distinguish between two or more parts of each type. The use of these terms is not intended to assign a particular order or orientation to the parts unless expressly stated otherwise.

[0039] As used herein, the term “antigen-binding molecule” refers to a polypeptide molecule (composed of one or more polypeptide chains) that can bind to an antigen. Antigen-binding molecules may originate from antibodies and typically contain an antigen-binding domain.

[0040] In this specification, the term "antibody" is used in its broadest sense and encompasses a variety of antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), heavy chain antibodies, antibody fragments, and antigen-binding molecules, as long as they exhibit the desired antigen-binding activity.

[0041] As used herein, the terms “heavy chain antibody,” “heavy chain-only antibody,” and “HCAb” refer to antibodies lacking a light chain.

[0042] The terms "full-length antibody," "intact antibody," and "whole antibody" are used herein as interchangeable to refer to antibodies having a structure substantially similar to that of a native antibody.

[0043] An "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an intact antibody that binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, cross-Fab, Fab', Fab'-SH, F(ab')2, diabodies, linear antibodies, single-chain antibody molecules (e.g., scFv and scFab), single-domain antibodies, and multispecific antibodies formed from antibody fragments. For an overview of specific antibody fragments, see Hollinger and Hudson, Nature Biotechnology 23:1126-1136 (2005).

[0044] A "single-domain antibody" refers to an antibody fragment consisting of a single monomeric antibody variable domain, such as a VHH, nanobody, shark-derived VNAR, autonomous VH domain, or autonomous VL domain. A single-domain antibody provides an antigen-binding site that specifically binds to an epitope; that is, the antigen-binding site is formed only by a single-domain antibody.

[0045] "VHH," or "VHH domain," or "nanobody" refers to a single-domain antibody derived from the variable domain of heavy-chain antibodies from camels (e.g., camels, dromedaries, llamas, alpacas, etc.) (see Nguyen VKet al., 2000, The EMBO Journal, 19, 921-930; Muyldermans S., 2001, J Biotechnol., 74, 277-302; and Vanlandschoot P. et al., 2011, Antiviral Research 92, 389-407 for an overview). The antigen-binding site of VHH lacks a light-chain variable domain. The VHH domain can be humanized.

[0046] As used herein, the term “antigen-binding domain” refers to a portion that specifically binds to a target antigen. In particular, the term refers to the antigen-binding domain of an antibody, i.e., the portion containing a region that binds to and is complementary to part or all of the antigen. Therefore, in certain embodiments, the antigen-binding domain as used herein is the antigen-binding domain of an antibody. Such an antigen-binding domain may be provided by an antibody fragment, for example, a Fab molecule, a single-chain antibody molecule, or a single-domain antibody, such as a VHH domain.

[0047] The term "variable region" or "variable domain" refers to a domain in the heavy or light chain of an antibody that is involved in antibody-antigen binding. This term includes the VHH domain. The variable domains of the heavy and light chains of native antibodies (VH and VL, respectively) generally have similar structures, and each domain contains four conserved framework regions (FRs) and complementarity-determining regions (CDRs). See, for example, Kindt et al., Kuby Immunology, 6th ed., WH Freeman & Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity. Furthermore, antibodies that bind to a specific antigen may be isolated by screening a library of complementary VL or VH domains, respectively, using the VH or VL domain of the antibody that binds to the antigen. See, for example, Portolano et al., J. Immunol. 150:880-887 (1993); Clarkson et al., Nature 352:624-628 (1991). When used herein in combination with variable region sequences, “Kabat numbering” refers to the numbering system described by Kabat et al. Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0048] As used herein, the amino acid positions of all constant regions and domains in the heavy and light chains are numbered according to the Kabat numbering system described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), and are referred to herein as "Kabat numbering" or "Kabat numbering."

[0049] Specifically, the Kabat numbering system (see Kabat, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991), pp. 647-660) is used for the light chain constant domain CL of kappa and lambda isotypes, while the Kabat EU index numbering system (see pp. 661-723) is used for the heavy chain constant domains (CH1, hinge, CH2, and CH3), in which case it is further clarified herein by the terms "Kabat EU index numbering" or "Kabat EU index numbering."

[0050] As used herein, the terms “binding site” or “antigen-binding site” refer to the site of a binding molecule that provides interaction with an antigen, i.e., one or more amino acid residues. For example, the antigen-binding site of an antigen-binding domain includes amino acid residues from a complementarity-determining region (CDR). The antigen-binding site may also be provided by, for example, one or more variable domains (also called variable regions). In a single-domain antibody, the antigen-binding site is provided by a single variable domain. In contrast, in a Fab fragment, the antigen-binding site is provided by VH and VL domains.

[0051] As used herein, the terms “hypervariable region” or “HVR” refer to each region of the antigen-binding domain whose sequence is hypervariable and which determines antigen-binding specificity, such as the “complementarity-determining region” (CDR). Generally, a variable domain contains three CDRs. Therefore, antibodies containing VH and VL contain six CDRs; three in VH (HCDR1, HCDR2, HCDR3) and three in VL (LCDR1, LCDR2, LCDR3). Illustrative CDRs as used herein include: (a) Hypervariable loops occurring at amino acid residues 26-32 (L1), 50-52 (L2), 91-96 (L3), 26-32 (H1), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)); (b) CDRs occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and (c) Antigen contact occurring at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al. J.Mol.Biol.262:732-745 (1996)).

[0052] Unless otherwise specified, CDR is determined according to Kabat et al. cited above. Those skilled in the art will understand that the designation of CDR may also be determined according to Chothia, McCallum, or any other scientifically recognized nomenclature cited above.

[0053] "Framework" or "FR" refers to variable domain residues other than the complementarity-determining region (CDR). The variable domain FR generally consists of the following four FR domains: FR1, FR2, FR3, and FR4. Therefore, HVR and FR sequences generally appear in the following order in the VH and VHH domains (or VL): FR1-HCDR1(LCDR1)-FR2-HCDR2(LCDR2)-FR3-HCDR3(LCDR3)-FR4. Unless otherwise specified, CDR residues and other residues within the variable domains (e.g., FR residues) are numbered herein according to Kabat et al. cited above.

[0054] In this specification, the term “immunoglobulin molecule” refers to a protein having the structure of a naturally occurring antibody. For example, IgG-class immunoglobulins are heterotetrameric glycoproteins with a weight of approximately 150,000 daltons, composed of two disulfide-linked light chains and two heavy chains. From the N-terminus to the C-terminus, each heavy chain has a variable domain (VH), also called a variable heavy chain domain or heavy chain variable region, followed by three constant domains (CH1, CH2, and CH3), also called heavy chain constant regions. Similarly, from the N-terminus to the C-terminus, each light chain has a variable domain (VL), also called a variable light chain domain or light chain variable region, followed by a constant light chain (CL) domain, also called a light chain constant region. The heavy chain of an immunoglobulin can be assigned to one of five types called α(IgA), δ(IgD), ε(IgE), γ(IgG), or μ(IgM), some of which can be further divided into subtypes such as γ1(IgG1), γ2(IgG2), γ3(IgG3), γ4(IgG4), α1(IgA1), and α2(IgA2). The light chain of an immunoglobulin may be assigned to one of two types called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain. An immunoglobulin essentially consists of two Fab molecules and an Fc domain, which are linked via an immunoglobulin hinge region.

[0055] The “class” of an antibody or immunoglobulin refers to the type of constant domain or constant region present in the heavy chain of the antibody or immunoglobulin. Antibodies have five main classes: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into “subclasses” (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy chain constant domains corresponding to different classes of immunoglobulins are called α, δ, ε, γ, and μ, respectively.

[0056] A "Fab molecule," "Fab," "Fab fragment," or "Fab domain" refers to a protein consisting of the VH domain and CH1 domain of the immunoglobulin heavy chain ("Fab heavy chain") and the VL domain and CL domain of the immunoglobulin light chain ("Fab light chain").

[0057] A “cross-Fab molecule,” “cross-Fab,” or “crossover Fab molecule” refers to a Fab molecule in which either the variable or constant regions of the heavy and light chains are exchanged. The cross-Fab operation allows for two different chain compositions of a cross-Fab molecule. In one case, the variable regions of the heavy and light chains of the Fab are exchanged, meaning the cross-Fab molecule includes a peptide chain consisting of a light chain variable region (VL) and a heavy chain constant region (CH1) (where CH1 may be fused to an Fc domain), and a peptide chain consisting of a heavy chain variable region (VH) and a light chain constant region (CL). On the other hand, when the constant regions of the Fab heavy and light chains are exchanged, the cross-Fab molecule includes a peptide chain consisting of a heavy chain variable region (VH) and a light chain constant region (CL) (where CL may be fused to an Fc domain), and a peptide chain consisting of a light chain variable region (VL) and a heavy chain constant region (CH1).

[0058] The term "conventional Fab molecule" refers to a Fab molecule composed of a Fab heavy chain containing VH and CH1 domains and a Fab light chain containing VL and CL domains.

[0059] A "single-strand variable fragment" or "scFv" molecule is a fusion protein of the variable domains of the heavy (VH) and light (VL) chains of an antibody, linked by a linker. Specifically, the linker is a short polypeptide of 10-25 amino acids, usually rich in glycine for flexibility and serine or threonine for solubility, and can connect the N-terminus of the VH to the C-terminus of the VL, or vice versa. This protein retains the specificity of the original antibody despite the removal of the constant region and the introduction of the linker.

[0060] The term "multispecificity" means that a binding molecule (e.g., an antibody) can specifically bind to at least two different antigens. A multispecific binding molecule (e.g., an antibody) can be, for example, a bispecific binding molecule. Typically, a bispecific binding molecule contains two antigen-binding sites, each specific to a different antigen. In certain embodiments, a multispecific (e.g., bispecific) binding molecule can simultaneously bind to two antigens, particularly two antigens expressed on the same cell, adjacent cells, or cells within the same tissue.

[0061] As used herein, the term "valence" refers to the presence of a specific number of antigen-binding sites within the binding molecule. Therefore, the term "monovalent binding to an antigen" indicates the presence of one (and no more than one) antigen-specific antigen-binding sites within the binding molecule.

[0062] As used herein, the term “antigen” refers to a molecule, such as a protein, to which an antigen-binding molecule binds. Useful antigens may be found, for example, on the surface of tumor cells, tumor stromal cells, virus-infected cells, other diseased cells, immune cells, released in serum, and / or in the extracellular matrix (ECM). In certain embodiments, the antigen is a human protein.

[0063] As used herein, the term “epitope” refers to a site on an antigen to which an antigen-binding site binds. Epitopes can be formed from a stretch of consecutive amino acids or from a configuration consisting of different regions of discontinuous amino acids. Epitopes often contain chemically active surface groups of the antigen, such as amino acids, glycan side chains, phosphoryl or sulfonyl groups, and may have specific three-dimensional structural properties and / or specific charge properties. Two different antigen-binding domains capable of binding to the same antigen may bind to different epitopes of that antigen. Such different antigen-binding domains are said not to compete for binding if both antigen-binding domains can bind to the antigen simultaneously. In this case, the two different antigen-binding domains are non-competitive. Alternatively, antigen-binding domains may compete for binding, i.e., exhibit competitive binding. This may result from partially overlapping epitopes or from steric hindrance preventing simultaneous binding of the two antigen-binding domains. Assays for determining whether an antigen-binding domain exhibits non-competitive or competitive binding are well known in the art and include competitive binding analyses using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative antibody binding assay available in the art.

[0064] As used herein, “target antigen” refers to an antigen presented on the surface of a target cell, such as an antigen on a cell in a tumor, such as a cancer cell or a cell in the tumor stroma, or an antigen on a T cell.

[0065] As used herein, “tumor-associated antigen” refers to any antigen presented on the surface of cells in a tumor, such as cancer cells or cells in the tumor stroma. Specific tumor-associated antigens include CEA, FAP, Her2, Her3, or EGFR.

[0066] The term “fibroblast-activating protein (FAP),” also known as prolyl endopeptidase FAP or seplacase (EC3.4.21), means any native FAP derived from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats), unless otherwise specified. FAP is expressed on cancer-associated fibroblasts (CAFs) in tumor stroma. The term encompasses not only “full-length,” unprocessed FAP, but also any form of FAP resulting from intracellular processing. The term also encompasses naturally occurring variants of FAP, such as splice variants or allele variants. In one embodiment, the antigen-binding molecule of the present invention can specifically bind to human, mouse, and / or cynomolgus monkey FAP. The amino acid sequence of human FAP is shown in UniProt (www.uniprot.org) accession number Q12884 (version 149) or NCBI (www.ncbi.nlm.nih.gov / ) RefSeq NP_004451.2. The extracellular domain (ECD) of human FAP extends from amino acid position 26 to 760. The amino acid sequence of mouse FAP is shown in UniProt accession number P97321 (version 126) or NCBI RefSeq NP_032012.1. The extracellular domain (ECD) of mouse FAP extends from amino acid position 26 to 761. Preferably, the anti-FAP binding molecule binds to the extracellular domain of FAP. Exemplary anti-FAP binding molecules are described in international patent applications WO2012 / 020006A2 and WO2020 / 070041A1.

[0067] The term "carcinoembryonic antigen (CEA)," also known as carcinoembryonic antigen-associated cell adhesion molecule 5 (CEACAM5), refers, unless otherwise specified, to any native CEA derived from any vertebrate source, including primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and mammals such as rodents (e.g., mice and rats). The amino acid sequence of human CEA is shown in UniProt accession number P06731 (version 151). CEA has long been identified as a tumor-associated antigen (Gold and Freedman, J Exp Med., 121:439-462, 1965; Berinstein NL, J Clin Oncol., 20:2197-2207, 2002). Originally classified as a protein expressed only in fetal tissues, CEA has now been identified in several healthy adult tissues. These tissues are primarily epithelial in origin, including cells of the gastrointestinal, respiratory, and urogenital tracts, as well as cells of the colon, cervix, sweat glands, and prostate (Nap et al., Tumor Biol., 9(2-3):145-53, 1988; Nap et al., Cancer Res., 52(8):2329-23339, 1992). Epithelial tumors and their metastases contain CEA as a tumor-associated antigen. The presence of CEA itself does not necessarily mean transformation into cancer cells, but it indicates the distribution of CEA. In normal tissues, CEA is generally expressed on the apical surface of cells (Hammarstrom S., Semin Cancer Biol. 9(2):67-81(1999)), preventing it from reaching antibodies in the bloodstream. In contrast to normal tissue, CEA tends to be expressed across the entire surface of cancer cells (Hammarstrom S., Semin Cancer Biol. 9(2):67-81 (1999)). This change in expression pattern makes CEA more likely to bind to antibodies within cancer cells. Furthermore, CEA expression increases in cancer cells. In addition, increased CEA expression promotes increased cell-cell adhesion, which may lead to metastasis (Marshall J., Semin Oncol., 30(a Suppl. 8):30-6, 2003).The prevalence of CEA expression in various tumor entities is generally very high. In line with published data, independent analyses performed on tissue samples confirmed this high prevalence: approximately 95% in colorectal carcinoma (CRC), 90% in pancreatic cancer, 80% in gastric cancer, 60% in non-small cell lung cancer (NSCLC, co-expressing with HER3), and 40% in breast cancer. Lower expression rates were observed in small cell lung cancer and glioblastoma.

[0068] "HER2" (also known as erbB-2 or CD340) refers to any native HER2 from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses not only "full-length" unprocessed HER2, but also any form of HER2 resulting from intracellular processing. The term also encompasses naturally occurring variants of HER2, such as splice variants or allele variants. In one embodiment, HER2 is human HER2. The amino acid sequence of human HER2 is shown in UniProt (www.uniprot.org) entry number Q9UK79 (version 95).

[0069] "Epidermal growth factor receptor (EGFR)," also known as the proto-oncogene c-ErbB-1 or receptor tyrosine-protein kinase erbB-1, refers to any native EGFR from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats), unless otherwise specified. The amino acid sequence of human EGFR is shown in UniProt accession number P00533.

[0070] As used herein, "T cell antigen" refers to any antigen present on the surface of a T lymphocyte.

[0071] The term "PD-1," also known as CD279, PD1, or programmed cell death protein 1, refers to any native PD-1 from any vertebrate source, including mammals such as primates (e.g., humans), non-human primates (e.g., cynomolgus monkeys), and rodents (e.g., mice and rats), in particular human protein PD-1 having the amino acid sequence shown in UniProt (www.uniprot.org) accession number Q15116.

[0072] As used herein, the terms “interferon-gamma,” “IFNγ,” or “IFN-γ” refer to any native IFNγ of any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses unprocessed IFNγ as well as any form of IFNγ resulting from processing in cells. The term also encompasses naturally occurring variants of IFNγ, e.g., splice variants or allele variants.

[0073] As used herein, the terms “interleukin-2” or “IL-2” refer to any native IL-2 from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses unprocessed IL-2 and any form of IL-2 resulting from intracellular processing. The term also encompasses naturally occurring variants of IL-2, e.g., splice variants or allele variants.

[0074] Cytokine receptors are cell surface glycoproteins that specifically bind to cytokines and transmit their signals. Generally, cytokine receptors function as oligomeric complexes consisting of 2 to 4 receptor chains, also called subunits, which may be the same or different. Therefore, the term “cytokine receptor complex” refers to a cytokine receptor composed of at least two subunits.

[0075] The IFNγ receptor complex comprises the IFNγR1 subunit and the IFNγR2 subunit. The terms “interferon gamma receptor 1” or “IFNγR1,” also called CD119 (differentiation cluster 119) or interferon gamma receptor α chain (IFNγRα), refer to any native IFNγR1 from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses “full-length” unprocessed IFNγR1 and any form of IFNγR1 resulting from intracellular processing. The term also encompasses naturally occurring variants of IFNγR1, e.g., splice variants or allele variants. In certain embodiments, IFNγR1 is human IFNγR1.

[0076] The term “interferon-gamma receptor 2” or “IFNγR2,” also known as interferon-gamma receptor β chain (IFNγRβ), refers, unless otherwise specified, to any native IFNγR2 from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses “full-length” unprocessed IFNγR2 and any form of IFNγR2 resulting from intracellular processing. The term also encompasses naturally occurring variants of IFNγR2, e.g., splice variants or allele variants. In certain embodiments, IFNγR2 is human IFNγR2.

[0077] The terms “IL-2Rα” or “α-subunit of the IL-2 receptor,” also known as CD25, as used herein, refer to any native IL-2Rα from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats), unless otherwise specified. The term encompasses “full-length” unprocessed IL-2Rα and any form of IL-2Rα resulting from intracellular processing. The term also encompasses naturally occurring variants of IL-2Rα, e.g., splice variants or allele variants. In certain embodiments, IL-2Rα is human IL-2Rα.

[0078] The term “IL-2Rβ” or “the β-subunit of the IL-2 receptor,” also known as CD122 or p70, refers, unless otherwise specified, to any native IL-2Rβ from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses “full-length” unprocessed IL-2Rβ and any form of IL-2Rβ resulting from intracellular processing. The term also encompasses naturally occurring variants of IL-2Rβ, e.g., splice variants or allele variants. In certain embodiments, IL-2Rβ is human IL-2Rβ.

[0079] The term “IL-2Rγ” or “the γ subunit of the IL-2 receptor,” also known as the common cytokine receptor γ subunit, common γ chain, γc, or CD132, refers, unless otherwise specified, to any native IL-2Rγ from any vertebrate source, including mammals, e.g., primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses “full-length” unprocessed IL-2Rγ and any form of IL-2Rγ resulting from intracellular processing. The term also encompasses naturally occurring variants of IL-2Rγ, e.g., splice variants or allele variants. In certain embodiments, IL-2Rγ is human IL-2Rγ.

[0080] Different associations of the individual IL-2R subunits IL-2Rα, IL-2Rβ, and IL-2Rγ can produce three IL-2R forms with different affinities to IL-2. High-affinity IL-2R refers to the heterotrimeric form of IL-2R consisting of IL-2Rα, IL-2Rβ, and IL-2Rγ. Intermediate-affinity IL-2R refers to the heterodimeric form of IL-2R consisting of IL-2Rβ and IL-2Rγ. On the other hand, low-affinity IL-2R refers to the monomeric form of IL-2R consisting of IL-2Rα alone (for a review, see, for example, Olejniczak and Kasprzak, Med Sci Monit 14, RA179-189 (2008)).

[0081] As used herein, the term “IL-2 receptor complex” refers to either a high-affinity IL-2 receptor or an intermediate-affinity IL-2 receptor.

[0082] In this specification, the terms “Fc domain” or “Fc region” are used to define the C-terminal region of an immunoglobulin heavy chain that includes at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, antibodies produced by host cells may undergo post-translational cleavage of one or more amino acids from the C-terminus of the heavy chain, particularly one or two amino acids. Thus, by expression of certain nucleic acid molecules encoding a full-length heavy chain, antibodies produced by host cells may contain the full-length heavy chain or cleaved variants of the full-length heavy chain. This is the case when the last two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, numbered according to the Kabat EU index). Therefore, the C-terminal lysine (Lys447) or C-terminal glycine (Gly446) and lysine (Lys447) in the Fc region may or may not be present. In one embodiment, the heavy chain containing the Fc region (subunit) specified herein, as included in the binding molecule according to the present invention, includes an additional C-terminal lysine (K447, numbered according to the Kabat EU index). In one embodiment, the heavy chain containing the Fc region (subunit) specified herein, as included in the binding molecule according to the present invention, includes a C-terminal glycine residue (G446, numbered according to the Kabat EU index) but does not include a C-terminal lysine (Lys477). Unless otherwise specified herein, the numbering of amino acid residues within the Fc region or constant region follows the EU numbering system, also known as the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see also above).As used herein, the term "subunit" of an Fc domain means one of the two polypeptides that form the dimeric Fc domain, i.e., a polypeptide that contains the C-terminal constant region of an immunoglobulin heavy chain and is capable of stable self-assembly. For example, a subunit of an IgG Fc domain contains the IgG CH2 and IgG CH3 constant domains.

[0083] "Modifications that promote the association of the first and second subunits of the Fc domain" are manipulations of the peptide backbone or post-translational modifications of the Fc domain subunits that reduce or prevent the association of a peptide containing an Fc domain subunit with an identical polypeptide for homodimer formation. As used herein, association-promoting modifications preferably involve distinct modifications made to each of two Fc domain subunits that are desired to associate (i.e., the first and second subunits of the Fc domain), and these modifications are complementary to each other in order to promote the association of the two Fc domain subunits. For example, the association-promoting modifications may alter the structure or charge of one or both of the Fc domain subunits so that they perform a sterically or electrostatically desired association, respectively. Thus, (hetero)dimerization occurs between a polypeptide containing a first Fc domain subunit and a polypeptide containing a second Fc domain subunit, which may not be identical in the sense that the further components fused to each subunit (e.g., antigen-binding domains) are not the same. In one embodiment, the modification that facilitates the association of the first and second subunits of the Fc domain includes an amino acid mutation, specifically an amino acid substitution, within the Fc domain. In a particular embodiment, the modification that facilitates the association of the first and second subunits of the Fc domain includes separate amino acid mutations, specifically amino acid substitutions, in each of the two subunits of the Fc domain.

[0084] One heterodimerization approach known in the art is the so-called "knob-into-hole" technique, which is described in detail with several examples, for example, in International Publication No. 96 / 027011, Ridgway, JB, et al., Protein Eng. 9 (1996) 617-621; Merchant, AM, et al., Nat. Biotechnol. 16 (1998) 677-681, and International Publication No. 98 / 050431. In the "knob-into-hole" technique, specific amino acids on each CH3 domain are manipulated to create a projection ("knob") on one CH3 domain and a cavity ("hole") on the other CH3 domain within the interface formed between two CH3 domains in the tertiary structure of the antibody. In the tertiary structure of a multispecific antibody, the introduced projection of one CH3 domain can be positioned within the introduced cavity of the other CH3 domain.

[0085] The term "effector function" refers to the biological activity resulting from the Fc region of an antibody, which varies depending on the antibody isotype. Examples of antibody effector functions include C1q binding and complement-dependent cell-mediated cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); antibody-dependent cell-mediated phagocytosis (ADCP); cytokine secretion; immunoconjugate-mediated antigen uptake by antigen-presenting cells; downregulation of cell surface receptors (e.g., B cell receptors); and B cell activation.

[0086] An "activated Fc receptor" is an Fc receptor that, following the binding of the antibody's Fc domain, triggers a signaling event that stimulates receptor-hosting cells to perform effector functions. Examples of human activated Fc receptors include FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32), and FcαRI (CD89).

[0087] Antibody-dependent cell-mediated cytotoxicity (ADCC) is an immune mechanism in which immune effector cells lyse antibody-coated target cells. Target cells are cells to which antibodies or derivatives containing an Fc region specifically bind, typically via a protein moiety at the N-terminus of the Fc region. As used herein, the term “reduced ADCC” is defined as either a reduction in the number of target cells lysed at a given time by a given concentration of antibody in the culture medium surrounding the target cells via the ADCC mechanism as defined above, and / or an increase in the concentration of antibody in the culture medium surrounding the target cells required to achieve the lysis of a given number of target cells at a given time via the ADCC mechanism. ADCC reduction is compared to unmanipulated ADCC mediated by the same antibody produced by the same type of host cells using the same standard production, purification, formulation, and storage methods (known to those skilled in the art). For example, ADCC reduction mediated by an antibody containing an amino acid substitution in its Fc domain that reduces ADCC is compared to ADCC mediated by the same antibody that does not contain this amino acid substitution in its Fc domain. Appropriate assays for measuring ADCC are well known in the art (see, for example, PCT Publication No. 2006 / 082515 or PCT Publication No. 2012 / 130831).

[0088] "Reduced binding," for example, reduced binding to the Fc receptor, refers to a decrease in affinity for each interaction, as measured, for example, by SPR. For clarity, this term also includes a reduction in affinity to zero (or below the detection limit of the analytical method), i.e., complete termination of the interaction. Conversely, "increased binding" refers to an increase in binding affinity for each interaction.

[0089] "Affinity" refers to the sum of the non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" as used herein refers to the intrinsic binding affinity that reflects the 1:1 interaction between the members of a binding pair (e.g., an antibody and an antigen). The affinity of molecule X for its partner Y can generally be expressed by its dissociation constant (KD). Affinity can be measured by well-established methods known in the art, including those described herein. A preferred method for measuring affinity is surface plasmon resonance (SPR).

[0090] As used herein, the terms “to manipulate,” “to be manipulated,” and “to manipulate” are considered to include any manipulation or post-translational modification of the peptide backbone of naturally occurring or recombinant polypeptides or fragments thereof. Manipulation includes alteration of amino acid sequences, alteration of glycosylation patterns, or modification of the side chain groups of individual amino acids, and combinations thereof.

[0091] As used herein, the term “amino acid mutation” encompasses amino acid substitutions, deletions, insertions, and modifications. Any combination of substitutions, deletions, insertions, and modifications can be made to arrive at the final construct, insofar as the final construct has the desired characteristics, e.g., reduced binding to the Fc receptor or increased association with another peptide. Deletions and insertions of amino acid sequences include deletions and insertions of amino and / or carboxyl terminals of amino acids. The preferred amino acid mutation is amino acid substitution. For example, to alter the binding properties of the Fc region, non-conservative amino acid substitution, i.e., replacing one amino acid with another amino acid having different structural and / or chemical properties, is particularly preferred. Amino acid substitutions include substitutions with non-natural amino acids or substitutions with natural amino acid derivatives of 20 standard amino acids (e.g., 4-hydroxyproline, 3-methylhistidine, ornithine, homoserine, 5-hydroxylysine). Amino acid mutations can be induced using genetic or chemical methods well known in the art. Genetic methods may include site-directed mutagenesis, PCR, gene synthesis, etc. Methods other than genetic manipulation to alter the side chain groups of amino acids, such as chemical modifications, may also be useful. Various names can be used herein to indicate the same amino acid mutation. For example, a substitution of proline to glycine at position 329 of the Fc domain can be denoted as 329G, G329, G329, P329G, or Pro329Gly.

[0092] "Fused" means that components (e.g., the Fab molecule and the Fc domain subunit) are linked directly by peptide bonds or via one or more peptide linkers. The term "fusion site" refers to a site on one component where that component is fused to another. For example, the cytokine receptor binding domain may be fused at its C-terminus to the N-terminus of the VH domain of the target binding domain. Therefore, the fusion site on the cytokine receptor binding domain is the C-terminus, and the fusion site on the target binding domain is the N-terminus.

[0093] The "amino acid sequence identity percentage (%)" for a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to those in the reference polypeptide sequence, after aligning the sequences to obtain the maximum possible sequence identity and introducing gaps as necessary, without considering any conservative substitutions as part of the sequence identity. Alignment for determining the amino acid sequence identity percentage can be achieved in various ways within the scope of the art, for example, using publicly available computer software such as BLAST, BLAST-2, Clustal W, Megalign (DNASTAR) software, or FASTA program packages. Those skilled in the art can determine appropriate parameters for sequence alignment, including any algorithm necessary to achieve the maximum possible alignment over the full length of the sequences being compared. Alternatively, the percentage identity value can be generated using the sequence comparison computer program ALIGN-2. The ALIGN-2 sequence comparison computer program was created by Genentech, Inc., and its source code is filed in the user documentation of the US Copyright Office (Washington DC, 20559), registered under US Copyright Registration No. TXU510087, and published in International Publication No. 2001 / 007611.

[0094] Unless otherwise specified, for the purposes of this specification, amino acid sequence identity % values ​​are generated using the ggsearch program in FASTA package version 36.3.8c or later, along with the BLOSUM50 comparison matrix. The FASTA program package was created by WRPearson and DJLipman ("Improved Tools for Biological Sequence Analysis", PNAS 85(1988) 2444-2448), WRPearson ("Effective protein sequence comparison", MethEnzymol. 266(1996) 227-25 258) and Pearson et al. (Genomics 46(1997) 24-36) and is publicly available at www.fasta.bioch.virginia.edu / fasta_www2 / fasta_down.shtml or www.ebi.ac.uk / Tools / sss / fasta.

[0095] Alternatively, you can use the public server accessible at fasta.bioch.virginia.edu / fasta_www2 / index.cgi to perform a global alignment (not just a local one) and compare sequences using the ggsearch(global protein:protein) program with default options (BLOSUM50;open:-10;ext:-2;Ktup=2). The amino acid identity percentage is shown in the output alignment header.

[0096] Cytokine mimetic The inventors have found that a pair of antigen-binding domains (hereinafter referred to as cytokine receptor-binding domains) capable of binding to different cytokine receptor subunits of a cytokine receptor complex can act as a cytokine agonist, i.e., mimic naturally occurring cytokines. Cytokine agonists are also called cytokine mimes. This is demonstrated by combining a pair of cytokine receptor-binding domains capable of binding to different cytokine receptor subunits in a single antigen-binding molecule. The antibody format of these molecules may offer advantageous properties compared to natural or recombinant cytokines. To achieve conditional activation of cytokine receptors, the pair of cytokine receptor-binding domains was split into two different antigen-binding molecules, i.e., split molecules or split antibodies. These antigen-binding molecules further include antigen-binding domains (hereinafter referred to as target-binding domains) capable of binding to a target antigen. Target-dependent assembly of cytokine receptor-binding domain pairs allows the molecules to act as cytokine mimes. Using a target-dependent approach makes it even possible to direct agonist activity to a desired site, such as cells or tissues expressing the target. The target-binding domains of a molecule can simultaneously bind to the same target antigen; that is, they bind to different epitopes of the antigen and are non-competitive. Such a pair of target-binding domains is also called a biparatopic target-binding domain. The assembly of a cytokine receptor-binding domain by a biparatopic target-binding domain allows the cytokine receptor-binding domain to specifically target the site of interest, i.e., the cell or environment in which the target antigen is expressed, and act as a cytokine mimetic. Therefore, a split antigen-binding molecule pair is a biparatopic antigen-binding molecule pair.

[0097] Split biparatopic antigen-binding molecule pair In one embodiment, the present invention provides a pair of antigen-binding molecules that specifically bind to a target antigen, comprising: a) i) a first target-binding domain, ii) a first cytokine receptor-binding domain, and iii) an Fc domain; and a second antigen-binding molecule comprising i) a second target-binding domain, ii) a second cytokine receptor-binding domain, and iii) an Fc domain, wherein the first target-binding domain can bind to a first epitope of the target antigen, the second target-binding domain can bind to a second epitope of the target antigen, the first and second target-binding domains do not compete for binding to the target antigen; the first cytokine receptor-binding domain can bind to a first cytokine receptor subunit, and the second cytokine receptor-binding domain can bind to a second cytokine receptor subunit.

[0098] The antigen-binding molecule pairs described above and herein may incorporate any of the features described below, either individually or in combination, unless the context indicates otherwise.

[0099] Cytokine receptor binding domain According to the present invention, each antigen-binding molecule in an antigen-binding molecule pair includes a cytokine receptor-binding domain. The first antigen-binding molecule includes a first cytokine receptor-binding domain, and the second antigen-binding molecule includes a second cytokine receptor-binding domain. The cytokine receptor-binding domains contained in the antigen-binding molecule pair bind to different cytokine receptor subunits. Each cytokine receptor-binding domain in the antigen-binding molecule pair binds to different cytokine receptor subunits of the cytokine receptor complex. Therefore, the first antigen-binding molecule includes a first cytokine receptor-binding domain that can bind to a first cytokine receptor subunit, and the second antigen-binding molecule includes a second cytokine receptor-binding domain that can bind to a second cytokine receptor subunit. The first and second cytokine receptor subunits are subunits of the cytokine receptor complex.

[0100] The cytokine receptor complex may be an IFNγ receptor complex, an IL-2 receptor complex, an IL-7 receptor complex, an IL-12 receptor complex, or an IL-18 receptor complex. Therefore, the first and second cytokine receptor binding domains may bind to different subunits of the IFNγ receptor complex, IL-2 receptor complex, IL-7 receptor complex, IL-12 receptor complex, or IL-18 receptor complex. In one embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain bind to a subunit of the IFNγ receptor complex. Thus, in one embodiment, the first cytokine receptor binding domain may bind to IFNγR1 and the second cytokine receptor binding domain may bind to IFNγR2. Alternatively, the first cytokine receptor binding domain may bind to IFNγR2 and the second cytokine receptor binding domain may bind to IFNγR1. In further embodiments, the first cytokine receptor binding domain and the second cytokine receptor binding domain bind to a subunit of the IL-2 receptor complex. Therefore, in one embodiment, the first cytokine receptor binding domain can bind to IL-2Rβ and the second cytokine receptor binding domain can bind to IL-2Rγ. In another embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain bind to a subunit of the IL-7 receptor complex. Therefore, in one embodiment, the first cytokine receptor binding domain can bind to IL-7Rα and the second cytokine receptor binding domain can bind to IL-2Rγ. In yet another embodiment, the first cytokine receptor binding domain and the second cytokine receptor binding domain bind to a subunit of the IL-12 receptor complex.Therefore, in one embodiment, the first cytokine receptor binding domain can bind to IL-12Rβ1, and the second cytokine receptor binding domain can bind to IL-12Rβ2. In another embodiment, the first cytokine receptor binding domain can bind to IL-12Rβ2, and the second cytokine receptor binding domain can bind to IL-12Rβ1.

[0101] The first and / or second cytokine receptor binding domains may be antibody fragments such as Fv, Fab, scFv, scFab, or single-domain antibodies. In one embodiment, the first and / or second cytokine receptor binding domains are single-domain antibodies. In a particular embodiment, the first and second cytokine receptor binding domains are VHH domains.

[0102] In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10. In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 3, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 7. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 7, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 3.

[0103] In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, and SEQ ID NO: 60, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65. In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, and SEQ ID NO: 60. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 60, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 60.

[0104] In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NOs: 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, and 192, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NOs: 61, 62, 63, 64, and 65. In one embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NOs. 61, 62, 63, 64, and 65, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NOs. 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, and 192. In another embodiment, the first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NOs. 177, 178, 179, 181, 182, 183, 186, 187, 189, 190, 191, and 192, and the second cytokine receptor binding domain includes the sequence of SEQ ID NOs. 62. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 177, SEQ ID NO: 178, SEQ ID NO: 179, SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 189, SEQ ID NO: 190, SEQ ID NO: 191, and SEQ ID NO: 192. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 182, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62. In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 182.In one embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 186, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62. In another embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 186. In yet another embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 190, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62. In yet another embodiment, the first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 190.

[0105] Target antigen binding domain According to the present invention, each antigen-binding molecule in an antigen-binding molecule pair includes a target-binding domain. The first antigen-binding molecule includes a first target-binding domain, and the second antigen-binding molecule includes a second target-binding domain. Both the first and second target-binding domains bind to the same target, i.e., the same antigen. However, the first and second target-binding domains bind to different epitopes of the target. The first target-binding domain can bind to a first epitope on the target antigen, and the second target-binding domain can bind to a second epitope on the target antigen. The first and second target-binding domains can bind to the target simultaneously; that is, the target-binding domains are non-competitive domains. The first and second target-binding domains do not compete for binding to the target antigen.

[0106] The target-binding domains of the first antigen-binding molecule and / or the second antigen-binding molecule may be antibody fragments. In one embodiment, the first target-binding domain is an antibody fragment, and the second target-binding domain is an antibody fragment. In one embodiment, the first antigen-binding molecule includes a first target-binding domain, the target-binding domain being an antibody fragment, and the second antigen-binding molecule includes a second target-binding domain, the target-binding domain being an antibody fragment. The target-binding domains of the first antigen-binding molecule and / or the second antigen-binding molecule may be Fv, Fab, scFv, scFab molecules, or single-domain antibodies. In one embodiment, the first antigen-binding molecule includes a first target-binding domain, the target-binding domain being an Fv, Fab, scFv, scFab molecule, or single-domain antibody, and the second antigen-binding molecule includes a second target-binding domain, the target-binding domain being an Fv, Fab, scFv, scFab molecule, or single-domain antibody.

[0107] In one embodiment, the target-binding domain of the first antigen-binding molecule and / or the second antigen-binding molecule is a Fab molecule. In one embodiment, the first target-binding domain is a Fab molecule, and the second target-binding domain is a Fab molecule. In one embodiment, the first antigen-binding molecule includes a first target-binding domain, the target-binding domain is a Fab molecule, and the second antigen-binding molecule includes a second target-binding domain, the target-binding domain is a Fab molecule. In one embodiment, the first target-binding domain includes a heavy-chain variable domain (VH1), a light-chain variable domain (VL1), a heavy-chain constant domain (CH11), and a light-chain constant domain (CL1). In one embodiment, the second target-binding domain includes a heavy-chain variable domain (VH2), a light-chain variable domain (VL2), a heavy-chain constant domain (CH12), and a light-chain constant domain (CL2).

[0108] The target-binding domain can be cross-Fab. In one embodiment, the first target-binding domain is cross-Fab. In another embodiment, the second target-binding domain is cross-Fab. In one embodiment, the first antigen-binding molecule includes the first target-binding domain, the target-binding domain is a Fab molecule, and the Fab molecule is cross-Fab; the second antigen-binding molecule includes the second target-binding domain, the target-binding domain is a Fab molecule, and the Fab molecule is not cross-Fab, i.e., a conventional Fab molecule. In another embodiment, the first antigen-binding molecule includes the first target-binding domain, the target-binding domain is a Fab molecule, and the Fab molecule is a conventional Fab molecule; the second antigen-binding molecule includes the second target-binding domain, the target-binding domain is a Fab molecule, and the Fab molecule is cross-Fab.

[0109] Both the first and second target-binding domains specifically bind to the same target antigen. The target antigen may be a tumor-associated antigen or a T-cell antigen.

[0110] The target antigen may be a tumor-associated antigen such as CEA, FAP, Her2, Her3, or EGFR. The target antigen may be a human tumor-associated antigen. Therefore, the target antigen may be human CEA, human FAP, human Her2, human Her3, or human EGFR. Thus, in one embodiment, the first target-binding domain can bind to a first epitope of CEA, and the second target-binding domain can bind to a second epitope of CEA. In one embodiment, the first target-binding domain can bind to a first epitope of FAP, and the second target-binding domain can bind to a second epitope of FAP. In one embodiment, the first target-binding domain can bind to a first epitope of Her2, and the second target-binding domain can bind to a second epitope of Her2. In one embodiment, the first target-binding domain can bind to a first epitope of Her3, and the second target-binding domain can bind to a second epitope of Her3. In one embodiment, the first target-binding domain can bind to a first epitope of EGFR, and the second target-binding domain can bind to a second epitope of EGFR.

[0111] In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 20 and VL1 of SEQ ID NO: 21, and the second target-binding domain includes VH2 of SEQ ID NO: 22 and VL2 of SEQ ID NO: 23. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 22 and VL1 of SEQ ID NO: 23, and the second target-binding domain includes VH2 of SEQ ID NO: 20 and VL2 of SEQ ID NO: 21. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 114 and VL1 of SEQ ID NO: 115, and the second target-binding domain includes VH2 of SEQ ID NO: 116 and VL2 of SEQ ID NO: 117. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 116 and VL1 of SEQ ID NO: 117, and the second target-binding domain includes VH2 of SEQ ID NO: 114 and VL2 of SEQ ID NO: 115. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 130 and VL1 of SEQ ID NO: 131, and the second target-binding domain includes VH2 of SEQ ID NO: 132 and VL2 of SEQ ID NO: 133. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 132 and VL1 of SEQ ID NO: 133, and the second target-binding domain includes VH2 of SEQ ID NO: 130 and VL2 of SEQ ID NO: 131. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 140 and VL1 of SEQ ID NO: 141, and the second target-binding domain includes VH2 of SEQ ID NO: 142 and VL2 of SEQ ID NO: 143. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 142 and VL1 of SEQ ID NO: 143, and the second target-binding domain includes VH2 of SEQ ID NO: 140 and VL2 of SEQ ID NO: 141.

[0112] The target antigen may be a T cell antigen such as PD-1 or LAG-3. The target antigen may be human PD-1 or human LAG-3. In one embodiment, the first target-binding domain can bind to a first epitope of PD-1, and the second target-binding domain can bind to a second epitope of PD-1. In one embodiment, the first target-binding domain can bind to a first epitope of LAG-3, and the second target-binding domain can bind to a second epitope of LAG-3. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 76 and VL1 of SEQ ID NO: 77, and the second target-binding domain includes VH2 of SEQ ID NO: 78 and VL2 of SEQ ID NO: 79. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 78 and VL1 of SEQ ID NO: 79, and the second target-binding domain includes VH2 of SEQ ID NO: 76 and VL2 of SEQ ID NO: 77. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 93 and VL1 of SEQ ID NO: 94, and the second target-binding domain includes VH2 of SEQ ID NO: 78 and VL2 of SEQ ID NO: 79. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 78 and VL1 of SEQ ID NO: 79, and the second target-binding domain includes VH2 of SEQ ID NO: 93 and VL2 of SEQ ID NO: 94. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 150 and VL1 of SEQ ID NO: 151, and the second target-binding domain includes VH2 of SEQ ID NO: 152 and VL2 of SEQ ID NO: 153. In one embodiment, the first target-binding domain includes VH1 of SEQ ID NO: 152 and VL1 of SEQ ID NO: 153, and the second target-binding domain includes VH2 of SEQ ID NO: 150 and VL2 of SEQ ID NO: 151.

[0113] FC Domain According to the present invention, the first antigen-binding molecule and the second antigen-binding molecule contain an Fc domain.

[0114] The Fc domain of the binding molecule consists of a polypeptide chain pair, a first Fc domain subunit, and a second Fc domain subunit. The first and second Fc domain subunits may contain the heavy chain domains of the immunoglobulin molecule. For example, the Fc domain of the immunoglobulin G (IgG) molecule is a dimer, and each of its subunits contains the CH2 and CH3 IgG heavy chain constant domains. The two subunits of the Fc domain can stably associate with each other.

[0115] In one embodiment, the first antigen-binding molecule includes an Fc domain composed of first and second Fc domain subunits, and / or the second antigen-binding molecule includes an Fc domain composed of first and second Fc domain subunits. In one embodiment, the first antigen-binding molecule includes an Fc domain composed of first and second Fc domain subunits, and the second antigen-binding molecule includes an Fc domain composed of first and second Fc domain subunits. In one embodiment, each antigen-binding molecule includes not more than one Fc domain.

[0116] In one embodiment, the Fc domain of the first and / or second antigen-binding molecule may be an IgG Fc domain. In a particular embodiment, the Fc domain is an IgG1Fc domain. In another particular embodiment, the Fc domain is an IgG4Fc domain.

[0117] In a more specific embodiment, the Fc domain is an IgG4Fc domain containing an amino acid substitution at position S228 (Kabat EU index numbering), particularly the amino acid substitution S228P. This amino acid substitution reduces Fab arm exchange of IgG4 antibodies in vivo (see Stubenrauch et al., Drug Metabolism and Disposition 38, 84-91 (2010)). In a further embodiment, the Fc domain is a human Fc domain. In a particular embodiment, the Fc domain is a human IgG1Fc domain.

[0118] In one embodiment, the Fc domain includes modifications that promote the association of the first and second subunits of the Fc domain. Further Fc domain modifications that promote heterodimerization are described below.

[0119] In one embodiment, the Fc domain includes one or more amino acid substitutions that reduce Fc receptor binding and / or effector function. Further Fc domain modifications that reduce Fc receptor binding and / or effector function are described below.

[0120] Fc domain modification that promotes heterodimerization The antigen-binding molecule according to the present invention comprises a target-binding domain and a cytokine receptor-binding domain that can be fused to a first and / or second Fc domain subunit, and thus the two Fc domain subunits are typically contained in two non-identical polypeptide chains. Recombinant co-expression of these polypeptides and subsequent dimerization result in several possible combinations of the two polypeptides. To increase the yield and purity of the antigen-binding molecule in recombinant production, it is advantageous to introduce modifications to the Fc domain of the antigen-binding molecule that promote the association of the desired polypeptide.

[0121] Therefore, in certain embodiments, the Fc domain of the antigen-binding molecule according to the present invention includes modifications that facilitate the association of the first and second subunits of the Fc domain. The site of the longest protein-protein interaction between the two subunits of the human IgG Fc domain is located within the CH3 domain of the Fc domain. Therefore, in one embodiment, the modification is located within the CH3 domain of the Fc domain.

[0122] To enhance heterodimerization, several approaches exist for modifying the CH3 domain of the Fc domain, which are well described, for example, in International Publications 96 / 27011, 98 / 050431, European Patent No. 1870459, 2007 / 110205, 2007 / 147901, 2009 / 089004, 2010 / 129304, 2011 / 90754, 2011 / 143545, 2012058768, 2013157954, and 2013096291. Typically, in all such approaches, the CH3 domain of the first subunit of the Fc domain and the CH3 domain of the second subunit of the Fc domain are both designed complementaryly so that each CH3 domain (or the heavy chain that comprises it) cannot homodimerize on its own, but is forced to heterodimerize with other complementaryly engineered CH3 domains (so that the first and second CH3 domains heterodimerize and homodimers are not formed between two first or two second CH3 domains).

[0123] In a specific embodiment, the modification that facilitates the association between the first and second subunits of the Fc domain is a so-called "knob-into-hole" modification, which includes a "knob" modification on one of the two subunits of the Fc domain and a "hole" modification on the other of the two subunits of the Fc domain.

[0124] The knob-into-hole technique is described, for example, in U.S. Patents 5,731,168, 7,695,936, Ridgway et al., Prot Eng 9,617-621 (1996), and Carter, J Immunol Meth 248,7-15 (2001). Generally, this method involves introducing a projection ("knob") at the interface of a first polypeptide and a corresponding cavity ("hole") at the interface of a second polypeptide, so that the projection can be positioned within the cavity to promote heterodimerization and inhibit homodimerization. The projection is constructed by replacing a small amino acid side chain from the interface of the first polypeptide with a larger side chain (e.g., tyrosine or tryptophan).

[0125] A compensatory cavity of the same or similar size as the protrusion is formed at the interface of the second polypeptide by replacing a larger amino acid side chain with a smaller amino acid side chain (e.g., alanine or threonine).

[0126] Therefore, in a preferred embodiment, in the CH3 domain of the first subunit of the Fc domain of the antigen-binding molecule, an amino acid residue is replaced with an amino acid residue having a larger side chain volume, thereby generating a repositionable protrusion within the CH3 domain of the first subunit within the cavity in the CH3 domain of the second subunit, and in the CH3 domain of the second subunit of the Fc domain, an amino acid residue is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the CH3 domain of the second subunit, within which the protrusion within the CH3 domain of the first subunit is repositionable.

[0127] Preferably, the amino acid residue having a larger side chain volume is selected from the group consisting of arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W). Preferably, the amino acid residue having a smaller side chain volume is selected from the group consisting of alanine (A), serine (S), threonine (T), and valine (V). The protrusions and cavities can be created by altering the nucleic acid encoding the polypeptide, for example, by site-directed mutagenesis or by peptide synthesis.

[0128] In a specific embodiment, in the CH3 domain of the first subunit ("knob" subunit) of the Fc domain, the threonine residue at position 366 is replaced with a tryptophan residue (T366W), and in the CH3 domain of the second subunit ("whole" subunit) of the Fc domain, the tyrosine residue at position 407 is replaced with a valine residue (Y407V). In another embodiment, in the second subunit of the Fc domain, the threonine residue at position 366 is further replaced with a serine residue (T366S), and the leucine residue at position 368 is replaced with an alanine residue (L368A) (numbered according to the Kabat EU index).

[0129] In further embodiments, the first subunit of the Fc domain may be further modified by replacing the serine residue at position 354 with a cysteine ​​residue (S354C) or the glutamic acid residue at position 356 with a cysteine ​​residue (E356C) (particularly the replacement of the serine residue at position 354 with a cysteine ​​residue), or the second subunit of the Fc domain may be further modified by replacing the tyrosine residue at position 349 with a cysteine ​​residue (Y349C) (numbered according to the Kabat EU index). The introduction of these two cysteine ​​residues leads to the formation of a disulfide bridge between the two subunits of the Fc domain, further stabilizing the dimer (Carter, J Immunol Methods 248, 7-15 (2001)).

[0130] In certain embodiments, the first subunit of the Fc domain contains amino acid substitutions S354C and T366W, and the second subunit of the Fc domain contains amino acid substitutions Y349C, T366S, L368A and Y407V (numbered according to the Kabat EU index).

[0131] Other techniques for CH3 modification that enhance heterodimerization have been considered as alternatives to the present invention and are described, for example, in International Publication Nos. 96 / 27011, 98 / 050431, European Patent No. 1870459, International Publication Nos. 2007 / 110205, 2007 / 147901, 2009 / 089004, 2010 / 129304, 2011 / 90754, 2011 / 143545, 2012 / 058768, 2013 / 157954, and 2013 / 096291.

[0132] In one embodiment, the heterodimerization approach described in European Patent No. 1870459 is used instead. This approach is based on the introduction of oppositely charged amino acids to specific amino acid positions at the CH3 / CH3 domain interface between two subunits of the Fc domain.

[0133] Specific embodiments of the antigen-binding molecule of the present invention are amino acid mutation R409D; K370E in one of the two CH3 domains (of the Fc domain) and amino acid mutation D399K; E357K in the other CH3 domain of the Fc domain (numbered according to the Kabat EU index).

[0134] In one embodiment, the antigen-binding molecule of the present invention contains the amino acid mutation T366W in the CH3 domain of the first subunit of the Fc domain, the amino acid mutations T366S, L368A, and Y407V in the CH3 domain of the second subunit of the Fc domain, and further amino acid mutations R409D and K370E in the CH3 domain of the first subunit of the Fc domain, and amino acid mutations D399K and E357K in the CH3 domain of the second subunit of the Fc domain (numbered according to the Kabat EU index).

[0135] In one embodiment, the antigen-binding molecule of the present invention contains amino acid mutations S354C and T366W in the CH3 domain of the first subunit of the Fc domain, and amino acid mutations Y349C, T366S, L368A, and Y407V in the CH3 domain of the second subunit of the Fc domain, or the antigen-binding molecule contains amino acid mutations Y349C and T366W in the CH3 domain of the first subunit of the Fc domain, amino acid mutations S354C, T366S, L368A, and Y407V in the CH3 domain of the second subunit of the Fc domain, and further contains amino acid mutations R409D and K370E in the CH3 domain of the first subunit of the Fc domain, and amino acid mutations D399K and E357K in the CH3 domain of the second subunit of the Fc domain (all numbered according to the Kabat EU index).

[0136] In one embodiment, the heterodimerization approach described in International Publication No. 2013 / 157953 is used instead. In one embodiment, the first CH3 domain contains the amino acid mutation T366K, and the second CH3 domain contains the amino acid mutation L351D (numbered according to the Kabat EU index). In a further embodiment, the first CH3 domain contains a further amino acid mutation L351K. In a further embodiment, the second CH3 domain further contains amino acid mutations selected from Y349E, Y349D, and L368E (particularly L368E) (numbered according to the Kabat EU index).

[0137] In one embodiment, the heterodimerization approach described in International Publication No. 2012 / 058768 is used instead. In one embodiment, the first CH3 domain contains amino acid mutations L351Y, Y407A, and the second CH3 domain contains amino acid mutations T366A, K409F. In a further embodiment, the second CH3 domain contains further amino acid mutations at positions T411, D399, S400, F405, N390, or K392, e.g., a) T411N, T411R, T411Q, T411K, T411D, T411E, or T411W, b) D399R, D399W, D399Y The amino acid mutations include D399K, c) S400E, S400D, S400R or S400K, d) F405I, F405M, F405T, F405S, F405V or F405W, e) N390R, N390K or N390D, f) K392V, K392M, K392R, K392L, K392F or K392E (numbered according to the Kabat EU index). In a further embodiment, the first CH3 domain includes amino acid mutations L351Y, Y407A, and the second CH3 domain includes amino acid mutations T366V, K409F. In a further embodiment, the first CH3 domain includes amino acid mutation Y407A, and the second CH3 domain includes amino acid mutations T366A, K409F. In a further embodiment, the second CH3 domain further includes amino acid mutations K392E, T411E, D399R, and S400R (numbered according to the Kabat EU index).

[0138] In one embodiment, the heterodimerization approach described in International Publication No. 2011 / 143545 is used alternatively by amino acid modification at a position selected from the group consisting of, for example, 368 and 409 (numbered according to the Kabat EU index).

[0139] In one embodiment, the heterodimerization approach described in International Publication No. 2011 / 090762, which also uses the knob-into-hole technique described above, is used as an alternative. In one embodiment, the first CH3 domain contains the amino acid mutation T366W and the second CH3 domain contains the amino acid mutation Y407A. In another embodiment, the first CH3 domain contains the amino acid mutation T366Y and the second CH3 domain contains the amino acid mutation Y407T (numbered according to the Kabat EU index).

[0140] In one embodiment, the antigen-binding molecule or its Fc domain is an IgG2 subclass, and the heterodimerization approach described in WO 2010 / 129304 is used as an alternative.

[0141] In alternative embodiments, modifications that facilitate the association of the first and second subunits of the Fc domain include modifications that mediate an electrostatic steering effect, such as those described in International Publication No. 2009 / 089004. Generally, this method involves the substitution of one or more amino acid residues with charged amino acid residues at the interface of the two Fc domain subunits such that homodimerization is electrostatically undesirable, but heterodimerization is electrostatically desirable. In some such embodiments, the first CH3 domain includes an amino acid substitution of K392 or N392 with a negatively charged amino acid (e.g., glutamic acid (E) or aspartic acid (D), particularly K392D or N392D), and the second CH3 domain includes an amino acid substitution of D399, E356, D356, or E357 with a positively charged amino acid (e.g., lysine (K) or arginine (R), particularly D399K, E356K, D356K, or E357K, more specifically D399K and E356K). In further embodiments, the first CH3 domain further includes an amino acid substitution of K409 or R409 with a negatively charged amino acid (e.g., glutamic acid (E) or aspartic acid (D), particularly K409D or R409D). In a further embodiment, the first CH3 domain further or alternatively comprises amino acid substitutions of K439 and / or K370 with negatively charged amino acids (e.g., glutamic acid (E) or aspartic acid (D)) (all numbering is by Kabat EU index).

[0142] In one embodiment, the heterodimerization approach described in International Publication No. 2007 / 147901 is used instead. In one embodiment, the first CH3 domain contains amino acid mutations K253E, D282K, and K322D, and the second CH3 domain contains amino acid mutations D239K, E240K, and K292D (numbered according to the Kabat EU index). In one embodiment, the heterodimerization approach described in International Publication No. 2007 / 110205 may be used as an alternative.

[0143] In one embodiment, the first subunit of the Fc domain contains amino acid substitutions K392D and K409D, and the second subunit of the Fc domain contains amino acid substitutions D356K and D399K (numbered according to the Kabat EU index).

[0144] Fc domain modification that reduces Fc receptor binding and / or effector function. The Fc domain confers favorable pharmacokinetic properties to antigen-binding molecules, including a long serum half-life that contributes to good accumulation in target tissues and a favorable tissue-to-blood distribution ratio. However, at the same time, it can lead to undesirable targeting of binding molecules to cells expressing the Fc receptor rather than the desired antigen-containing cells. Furthermore, co-activation of the Fc receptor signaling pathway can lead to cytokine release, which can result in hyperactivation of cytokine receptors and serious side effects upon systemic administration. Activation of immune cells other than T cells (that possess the Fc receptor) can even reduce the effectiveness of antigen-binding molecule pairs, for example, due to the potential destruction of T cells by NK cells.

[0145] Therefore, in certain embodiments, the Fc domain of the antigen-binding molecule according to the present invention exhibits reduced binding affinity to the Fc receptor and / or reduced effector function compared to the native IgG1Fc domain. In some such embodiments, the Fc domain (or the antigen-binding molecule containing said Fc domain) exhibits a binding affinity to the Fc receptor of less than 50%, specifically less than 20%, more specifically less than 10%, and most specifically less than 5%, compared to the native IgG1Fc domain (or the antigen-binding molecule containing the native IgG1Fc domain), and / or an effector function of less than 50%, specifically less than 20%, more specifically less than 10%, and most specifically less than 5%, compared to the native IgG1Fc domain (or the antigen-binding molecule containing the native IgG1Fc domain). In one embodiment, the Fc domain (or the antigen-binding molecule containing said Fc domain) does not substantially bind to the Fc receptor and / or induce effector function. In certain embodiments, the Fc receptor is the Fcγ receptor. In one embodiment, the Fc receptor is the human Fc receptor. In one embodiment, the Fc receptor is an activated Fc receptor. In a specific embodiment, the Fc receptor is an activated human Fcγ receptor, more specifically human FcγRIIIa, FcγRI, or FcγRIIa, and most specifically human FcγRIIIa. In one embodiment, the effector function is one or more selected from the group of CDC, ADCC, ADCP, and cytokine secretion. In a specific embodiment, the effector function is ADCC. In one embodiment, the Fc domain exhibits substantially similar binding affinity to the neonatal Fc receptor (FcRn) compared to the native IgG1Fc domain domain. Substantially similar binding to FcRn is achieved when the Fc domain (or an antigen-binding molecule containing said Fc domain) exhibits a binding affinity to FcRn that is greater than approximately 70%, particularly greater than approximately 80%, and more specifically greater than approximately 90%, compared to the native IgG1Fc domain (or an antigen-binding molecule containing the native IgG1Fc domain).

[0146] In certain embodiments, the Fc domain is engineered to have reduced binding affinity to the Fc receptor and / or reduced effector function compared to an unengineered Fc domain. In certain embodiments, the Fc domain of an antigen-binding molecule contains one or more amino acid mutations that reduce the binding affinity and / or effector function of the Fc domain to the Fc receptor. Typically, the same one or more amino acid mutations are present in each of the two subunits of the Fc domain. In one embodiment, the amino acid mutations reduce the binding affinity of the Fc domain to the Fc receptor. In one embodiment, the amino acid mutations reduce the binding affinity of the Fc domain to the Fc receptor by at least half, at least one-fifth, or at least one-tenth. In embodiments where there is more than one amino acid mutation that reduces the binding affinity of the Fc domain to the Fc receptor, the combination of these amino acid mutations may reduce the binding affinity of the Fc domain to the Fc receptor by at least one-tenth, at least one-twentieth, or even at least one-fiftieth. In one embodiment, an antigen-binding molecule containing an engineered Fc domain exhibits a binding affinity to the Fc receptor of less than 20%, particularly less than 10%, and more specifically less than 5%, compared to an antigen-binding molecule containing an unengineered Fc domain. In a particular embodiment, the Fc receptor is an Fcγ receptor. In one embodiment, the Fc receptor is a human Fc receptor. In one embodiment, the Fc receptor is an activated Fc receptor. In a specific embodiment, the Fc receptor is an activated human Fcγ receptor, more specifically human FcγRIIIa, FcγRI, or FcγRIIa, and most specifically human FcγRIIIa. Preferably, binding to each of these receptors is reduced. In one embodiment, binding affinity to complement components, specifically to C1q, is also reduced. In one embodiment, binding affinity to the neonatal Fc receptor (FcRn) is not reduced.

[0147] Substantially similar binding to FcRn (i.e., protection of the binding affinity of the Fc domain to the receptor) is achieved when the Fc domain (or an antigen-binding molecule containing the Fc domain) exhibits a binding affinity greater than approximately 70% of the binding affinity of the unmodified form of the Fc domain (or an antigen-binding molecule containing this unmodified form of Fc) to FcRn. The Fc domain, or the antigen-binding molecule of the present invention containing the Fc domain, may exhibit an affinity greater than approximately 80% and even greater than approximately 90% of such affinity. In certain embodiments, the Fc domain of the antigen-binding molecule is modified to have reduced effector function compared to the unmodified Fc domain. Reduced effector function may include, but is not limited to, one or more of the following: reduced complement-dependent cell-mediated cytotoxicity (CDC), reduced antibody-dependent cell-mediated cytotoxicity (ADCC), reduced antibody-dependent cell phagocytosis (ADCP), reduced cytokine secretion, reduced immune complex-mediated antigen uptake by antigen-presenting cells, reduced binding to NK cells, reduced binding to macrophages, reduced binding to monocytes, reduced binding to polymorphonuclear cells, reduced direct signaling-induced apoptosis, reduced cross-linking with target-binding antibodies, reduced dendritic cell maturation, or reduced T cell priming. In one embodiment, reduced effector function is one or more selected from the group of reduced CDC, reduced ADCC, reduced ADCP, and reduced cytokine secretion. In a particular embodiment, reduced effector function is reduced ADCC. In one embodiment, reduced ADCC is less than 20% of ADCC induced by an unmanipulated Fc domain (or a binding molecule containing an unmanipulated Fc domain).

[0148] In one embodiment, the amino acid mutation that reduces the binding affinity and / or effector function of the Fc domain to the Fc receptor is an amino acid substitution. In one embodiment, the Fc domain contains an amino acid substitution at a position selected from the group E233, L234, L235, N297, P331, and P329 (numbered according to the Kabat EU index). In a more specific embodiment, the Fc domain contains an amino acid substitution at a position selected from the group L234, L235, and P329 (numbered according to the Kabat EU index). In one embodiment, the Fc domain contains the amino acid substitutions L234A and L235A (numbered according to the Kabat EU index). In some such embodiments, the Fc domain is an IgG1Fc domain, in particular a human IgG1Fc domain. In one embodiment, the Fc domain contains an amino acid substitution at position P329. In a more specific embodiment, the amino acid substitution is P329A or P329G, in particular P329G (numbered according to the Kabat EU index). In one embodiment, the Fc domain contains an amino acid substitution at position P329 and further amino acid substitutions at positions selected from E233, L234, L235, N297, and P331 (numbered according to the Kabat EU index). In a more specific embodiment, the further amino acid substitutions are E233P, L234A, L235A, L235E, N297A, N297D, or P331S. In a particular embodiment, the Fc domain contains amino acid substitutions at positions P329, L234, and L235 (numbered according to the Kabat EU index). In a more specific embodiment, the Fc domain contains amino acid mutations L234A, L235A, and P329G ("P329G LALA", "PGLALA", or "LALAPG").Specifically, in certain embodiments, each subunit of the Fc domain contains amino acid substitutions L234A, L235A, and P329G (Kabat EU index numbering), namely, in the first and second subunits of the Fc domain, the leucine residue at position 234 is replaced with an alanine residue (L234A), the leucine residue at position 235 is replaced with an alanine residue (L235A), and the proline residue at position 329 is replaced with a glycine residue (P329G) (Kabat EU index numbering).

[0149] In some such embodiments, the Fc domain is an IgG1Fc domain, particularly a human IgG1Fc domain. The amino acid substitution combination "P329G LALA" almost completely eliminates the Fcγ receptor (and complement) binding of the human IgG1Fc domain, as described in International Publication 2012 / 130831, which is incorporated herein in whole by reference. International Publication 2012 / 130831 also describes methods for preparing such mutant Fc domains and determining their properties (such as Fc receptor binding or effector function).

[0150] IgG4 antibodies exhibit reduced binding affinity to Fc receptors and reduced effector function compared to IgG1 antibodies. Therefore, in one embodiment, the Fc domain of the binding molecule of the present invention is an IgG4Fc domain, particularly a human IgG4Fc domain. In one embodiment, the IgG4Fc domain includes an amino acid substitution at position S228, specifically amino acid substitution S228P (numbered according to the Kabat EU index). To further reduce its binding affinity to the Fc receptor and / or its effector function, in one embodiment, the IgG4 Fc domain includes an amino acid substitution at position L235, specifically amino acid substitution L235E (numbered according to the Kabat EU index). In one embodiment, the IgG4Fc domain includes an amino acid substitution at position P329, specifically amino acid substitution P329G (numbered according to the Kabat EU index). In a preferred embodiment, the IgG4Fc domain includes amino acid substitutions at positions S228, L235, and P329, specifically amino acid substitutions S228P, L235E, and P329G (numbered according to the Kabat EU index). Such IgG4Fc domain variants and their Fcγ receptor binding properties are described in PCT Publication No. 2012 / 130831 (in whole, incorporated herein by reference).

[0151] In certain embodiments, Fc domains exhibiting reduced binding affinity to the Fc receptor and / or reduced effector function compared to native IgG1Fc domains are human IgG1Fc domains containing amino acid substitutions L234A, L235A and optionally P329G, or human IgG4Fc domains containing amino acid substitutions S228P, L235E and optionally P329G (numbered according to the Kabat EU index).

[0152] In certain embodiments, N-glycosylation of the Fc domain is excluded. In some such embodiments, the Fc domain contains amino acid mutations at position N297, particularly amino acid substitutions in which asparagine is replaced with alanine (N297A) or aspartic acid (N297D) (numbered according to the Kabat EU index).

[0153] In addition to the Fc domains described herein and in International Publication No. 2012 / 130831, Fc domains with reduced Fc receptor binding and / or effector function also include substitutions of one or more Fc domain residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent No. 6,737,056) (numbered by the Kabat EU Index). Such Fc variants include the so-called "DANA" Fc variant (U.S. Patent No. 7,332,581) in which residues 265 and 297 are substituted with alanine, and Fc variants having substitutions of two or more of the amino acid positions 265, 269, 270, 297, and 327.

[0154] Mutant Fc domains can be prepared by amino acid deletion, substitution, insertion, or modification using genetic or chemical methods well known in the art. Genetic methods include site-directed mutagenesis of the encoding DNA sequence, PCR, gene synthesis, etc. Correct nucleotide changes can be confirmed, for example, by sequencing.

[0155] Binding to the Fc receptor can be easily determined, for example, by ELISA, or by surface plasmon resonance (SPR) using Fc receptors obtained by recombinant expression with standard equipment such as a BIAcore instrument (GE Healthcare). Alternatively, the binding affinity of an Fc domain or a binding molecule containing an Fc domain to the Fc receptor may be evaluated using cell lines known to express a specific Fc receptor (e.g., human NK cells expressing the FcγIIIa receptor).

[0156] The effector function of the Fc domain, or a binding molecule containing an Fc domain, can be measured by methods known in the art. Examples of in vitro assays for evaluating the ADCC activity of a molecule of interest are described in U.S. Patent No. 5,500,362, Hellstrom et al., Proc Natl Acad Sci USA 83,7059-7063 (1986) and Hellstrom et al., Proc Natl Acad Sci USA 82,1499-1502 (1985), U.S. Patent No. 5,821,337, Bruggemann et al., J Exp Med 166,1351-1361 (1987).

[0157] Alternatively, non-radioactive assays may be used (see, for example, ACTI® non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc., Mountain View, CA); and CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, WI)). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells.

[0158] Alternatively, or in addition, the ADCC activity of the molecule in question may be evaluated in vivo, for example, in an animal model disclosed in Clynes et al., Proc Natl Acad Sci USA 95, 652-656 (1998).

[0159] In one embodiment, the binding of the Fc domain to complement components, particularly C1q, is reduced. Thus, in one embodiment in which the Fc domain is manipulated to have reduced effector function, this reduced effector function includes reduced CDC. C1q binding assays may be performed to determine whether an antigen-binding molecule containing an Fc domain or an Fc domain can bind to C1q and therefore possesses CDC activity. See, for example, the C1q and C3c binding ELISAs in International Publication Nos. 2006 / 029879 and International Publication Nos. 2005 / 100402. A CDC assay may be performed to evaluate complement activation (see, for example, Gazzano-Santoro et al., J Immunol Methods 202, 163 (1996); Cragg et al., Blood 101, 1045-1052 (2003); and Cragg and Glennie, Blood 103, 2738-2743 (2004)).

[0160] The determination of FcRn binding and in vivo clearance / half-life can also be carried out using methods known in the art (e.g., Petkova, S B et al., Int'l. Immunol. 18(12):1759-1769 (2006); see International Publication No. 2013 / 120929).

[0161] Structure of antigen-binding molecules The antigen-binding molecule according to the present invention may have various molecular configurations; that is, the domains of the binding molecule may be linked to each other in different ways.

[0162] The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, where the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the target-binding domain, and the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits. The target-binding domain may be a Fab molecule. Thus, the cytokine receptor-binding domain may be fused to the VH or VL domain of the target-binding domain. In one embodiment, the antigen-binding molecule includes a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, where the target-binding domain is a Fab molecule, the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the VH domain of the target-binding domain, and the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits (see, for example, Figures 11B, 11C, 11F, and 11G for schematic diagrams). In one embodiment, the antigen-binding molecule comprises a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain is a Fab molecule, the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the VL domain of the target-binding domain, and the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits (see, for example, Figures 11D, 11E, 11H, and 11I for schematic diagrams).

[0163] The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, where the target-binding domain is fused at its C-terminus to the N-terminus of a first Fc domain subunit, and the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of a second Fc domain subunit (see, for example, Figures 13A, 13B, 13C, and 13D for schematic diagrams).

[0164] The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, where the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits, and the cytokine receptor-binding domain is fused at its N-terminus to the C-terminus of the same Fc domain subunit (see, for example, Figures 13E, 13F, 13G, and 13H for schematic diagrams).

[0165] The cytokine receptor-binding domain can be fused via a peptide linker. The peptide linker may contain the amino acid sequence given by SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 279, SEQ ID NO: 280, or SEQ ID NO: 281.

[0166] antigen-binding molecule pair In one embodiment, the antigen-binding molecule of the antigen-binding molecule pair comprises a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the target-binding domain, and the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits.

[0167] In one embodiment, the target binding domain of the antigen-binding molecule is a Fab molecule. Thus, in one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a first cytokine receptor binding domain, VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VL1 and CL1 in order from the N-terminus to the C-terminus; and a second antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a second cytokine receptor binding domain, VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VL2 and CL2 in order from the N-terminus to the C-terminus.

[0168] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a first cytokine receptor binding domain, VL1 and CL1 in order from the N-terminus to the C-terminus; and a second antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus.

[0169] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing a first Fc domain subunit, a second polypeptide comprising a first cytokine receptor binding domain, VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VL1 and CL1 in order from the N-terminus to the C-terminus; and a second antigen-binding molecule comprising a first polypeptide containing a first Fc domain subunit, a second polypeptide comprising VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus.

[0170] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a first cytokine receptor binding domain, VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VH1 and CL1 in order from the N-terminus to the C-terminus, wherein the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a second cytokine receptor binding domain, VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VL2 and CL2 in order from the N-terminus to the C-terminus.

[0171] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a first cytokine receptor binding domain, VH1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus.

[0172] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a first cytokine receptor binding domain, VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VH1 and CL1 in order from the N-terminus to the C-terminus, wherein the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus.

[0173] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising VL1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising a first cytokine receptor binding domain, VH1 and CL1 in order from the N-terminus to the C-terminus; the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising a second cytokine receptor binding domain, VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide comprising VL2 and CL2 in order from the N-terminus to the C-terminus.

[0174] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 27, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 30, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0175] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 38, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0176] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33.

[0177] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 35, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36.

[0178] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 27, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33.

[0179] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 30, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31.

[0180] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 38, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36.

[0181] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 35.

[0182] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 260, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0183] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 119, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 256, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 122, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 255.

[0184] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0185] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 119, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 257, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 112, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 254.

[0186] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 260, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 112, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 255.

[0187] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 256, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0188] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 254.

[0189] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 257, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0190] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 86, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 83, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 87.

[0191] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 92, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 91.

[0192] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 95, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0193] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 99, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102.

[0194] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0195] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 100, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 104.

[0196] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 99, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0197] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 95, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102.

[0198] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 100, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0199] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 104.

[0200] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0201] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109.

[0202] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 111, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 112, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0203] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108.

[0204] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109.

[0205] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0206] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 111, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108.

[0207] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0208] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 125, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 129, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 127.

[0209] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 118, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 122, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 123.

[0210] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 126, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 128, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 127.

[0211] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 119, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 120, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 122, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 121.

[0212] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 125, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 122, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 123.

[0213] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 118, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 129, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127.

[0214] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 126, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 122, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 121.

[0215] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 119, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 120, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 128, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 127.

[0216] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 136, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 134, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 138, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 137.

[0217] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 135, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 134, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 138, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 139.

[0218] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 146, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 144, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 148, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 147.

[0219] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 145, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 144, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 148, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 149.

[0220] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163.

[0221] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159.

[0222] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 155, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 156, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 164, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 163.

[0223] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 162, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 160, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 158, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 157.

[0224] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 161, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 160, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 165, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 163.

[0225] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 155, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 154, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 106, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 158, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 159.

[0226] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163.

[0227] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157.

[0228] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 194, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 193, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0229] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 195, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0230] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 196, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0231] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 197, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0232] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 198, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0233] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 199, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0234] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 200, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0235] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 201, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0236] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 202, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0237] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 203, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0238] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 204, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0239] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 205, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89.

[0240] In one embodiment, both antigen-binding molecules of an antigen-binding molecule pair include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain is fused at its C-terminus to the N-terminus of a first Fc domain subunit, and the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of a second Fc domain subunit. In another embodiment, both antigen-binding molecules of an antigen-binding molecule pair include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain is fused at its C-terminus to the N-terminus of a first Fc domain subunit, and the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of a second Fc domain subunit, and the target-binding domain is a Fab molecule. In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising, in order from the N-terminus to the C-terminus, a first polypeptide comprising VH1, CH11 and a first Fc domain subunit; a second polypeptide comprising VL1 and CL1 in order from the N-terminus to the C-terminus; and a third polypeptide comprising a first cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus. The second antigen-binding molecule comprises a first polypeptide comprising VH2, CH12 and a first Fc domain subunit in order from the N-terminus to the C-terminus; a second polypeptide comprising VL2 and CL2 in order from the N-terminus to the C-terminus; and a third polypeptide comprising a second cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus.

[0241] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising, in order from the N-terminus to the C-terminus, a first polypeptide comprising VL1, CH11, and a first Fc domain subunit; a second polypeptide comprising VH1 and CL1, in order from the N-terminus to the C-terminus; and a third polypeptide comprising a first cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus; and a second antigen-binding molecule comprising, in order from the N-terminus to the C-terminus, a first polypeptide comprising VH2, CH12, and a first Fc domain subunit; a second polypeptide comprising VL2 and CL2, in order from the N-terminus to the C-terminus; and a third polypeptide comprising a second cytokine receptor-binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus.

[0242] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 32, a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 39, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 34, a second polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40.

[0243] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 34, a second polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 39, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 32, a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40.

[0244] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 238, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0245] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 236, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0246] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 239, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 240, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0247] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0248] In one aspect, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 236, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 32, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 235, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 34, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0249] In one aspect, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 240, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 32, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 239, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 34, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0250] In one aspect, both antigen-binding molecules of the antigen-binding molecule pair include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the target-binding domain is fused to the N-terminus of one of the Fc domain subunits at its C-terminus, and the cytokine receptor-binding domain is fused to the C-terminus of the same Fc domain subunit at its N-terminus.

[0251] Thus, in one aspect, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first target-binding domain fused to the N-terminus of one of the Fc domain subunits at its C-terminus and a first cytokine receptor-binding domain fused to the C-terminus of the same Fc domain subunit at its N-terminus, and a second antigen-binding molecule comprising a second target-binding domain fused to the N-terminus of one of the Fc domain subunits at its C-terminus and a second cytokine receptor-binding domain fused to the C-terminus of the same Fc domain subunit at its N-terminus.

[0252] In one embodiment, the target-binding domain is a Fab molecule. Therefore, in one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CH11 to the N-terminus of one of the Fc domain subunits, and a first cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit; and a second antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CH12 to the N-terminus of one of the Fc domain subunits, and a second cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit.

[0253] In one embodiment, one of the target-binding domains of the antigen-binding molecule is a cross-Fab molecule. Therefore, in one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CH11 to the N-terminus of one of the Fc domain subunits, and a first cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit; and a second antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CL2 to the N-terminus of one of the Fc domain subunits, and a second cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit. In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CL1 to the N-terminus of one of the Fc domain subunits, and a first cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit; and a second antigen-binding molecule comprising a target-binding domain fused at the C-terminus of its CH12 to the N-terminus of one of the Fc domain subunits, and a second cytokine receptor-binding domain fused at its N-terminus to the C-terminus of the same Fc domain subunit.

[0254] In one aspect, the pair of antigen-binding molecules includes a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising, in order from the N-terminus to the C-terminus, VH1, CH11, a second Fc domain subunit, and a first cytokine receptor-binding domain, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, VL1 and CL1; and a second antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising, in order from the N-terminus to the C-terminus, VH2, CH12, and a second Fc domain subunit and a second cytokine receptor-binding domain, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, VL2 and CL2.

[0255] In one aspect, the pair of antigen-binding molecules includes a first antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising, in order from the N-terminus to the C-terminus, VL1, CH11, a second Fc domain subunit, and a first cytokine receptor-binding domain, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, VH1 and CL1; and a second antigen-binding molecule comprising a first polypeptide comprising a first Fc domain subunit, a second polypeptide comprising, in order from the N-terminus to the C-terminus, VH2, CH12, and a second Fc domain subunit and a second cytokine receptor-binding domain, and a third polypeptide comprising, in order from the N-terminus to the C-terminus, VL2 and CL2.

[0256] In one aspect, the pair of antigen-binding molecules includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 42, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 41; and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 45, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 44, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 43.

[0257] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 46, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 41, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 47, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 43.

[0258] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 246, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 249, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0259] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 242, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 245, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0260] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 250, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 253, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0261] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 248, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 247, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0262] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 244, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 243, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0263] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 252, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 25, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 241, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 251, and a third polypeptide comprising the amino acid sequence of SEQ ID NO: 29.

[0264] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 245, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29.

[0265] Alternative configuration of antigen-binding molecules The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain, wherein the cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the target-binding domain, and the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits, and the target-binding domain is a VHH domain. Therefore, the antigen-binding molecule may include a first polypeptide containing a first Fc domain subunit and a second polypeptide containing a first cytokine receptor-binding domain, a first target-binding domain, and a second Fc domain subunit, in that order from the N-terminus to the C-terminus, and the second antigen-binding molecule may include a first polypeptide containing a first Fc domain subunit and a second polypeptide containing a second cytokine receptor-binding domain, a second target-binding domain, and a second Fc domain subunit, in that order from the N-terminus to the C-terminus.

[0266] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 173, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 171.

[0267] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 176, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 172.

[0268] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 175, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 171.

[0269] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 176, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 174.

[0270] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 175, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 170.

[0271] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 173, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 170.

[0272] In one embodiment, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 176, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 172.

[0273] The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain. The cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the target-binding domain. The target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits. The cytokine receptor-binding domain is a Fab molecule.

[0274] In one aspect, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 267, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 268, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 265, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 272, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 271.

[0275] In one aspect, the antigen-binding molecule pair includes a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 267, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 268, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 265, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 274, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 273.

[0276] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 269, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 268, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 272, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 271.

[0277] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 269, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 268, and a second antigen-binding molecule comprising a first polypeptide comprising the amino acid sequence of SEQ ID NO: 28, a second polypeptide comprising the amino acid sequence of SEQ ID NO: 274, a third polypeptide comprising the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide comprising the amino acid sequence of SEQ ID NO: 273.

[0278] The antigen-binding molecule may include a target-binding domain, a cytokine receptor-binding domain, and an Fc domain. The cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of the target-binding domain, the target-binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits, and the cytokine receptor-binding domain is an scFv domain.

[0279] In one embodiment, the antigen-binding molecule pair comprises a first antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 276, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 275, and a second antigen-binding molecule comprising a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 277, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 278.

[0280] Specific embodiments of the present invention The following lists specific embodiments of the present invention.

[0281] 1. an antigen-binding molecule pair that specifically binds to a target antigen, comprising: a) a first antigen-binding molecule comprising i) a first target-binding domain, ii) a first cytokine receptor-binding domain, and iii) an Fc domain; and b) a second antigen-binding molecule comprising i) a second target-binding domain, ii) a second cytokine receptor-binding domain, and iii) an Fc domain, wherein the first target-binding domain can bind to a first epitope on the target antigen, the second target-binding domain can bind to a second epitope on the target antigen, the first and second target-binding domains do not compete for binding to the target antigen, the first cytokine receptor-binding domain can bind to a first cytokine receptor subunit, and the second cytokine receptor-binding domain can bind to a second cytokine receptor subunit.

[0282] 2. The antigen-binding molecule pair according to Embodiment 1, wherein the first target-binding domain and the second target-binding domain are antibody fragments such as Fv, Fab, scFv, scFab molecules, or single-domain antibodies.

[0283] 3. The antigen-binding molecule pair according to Embodiment 1 or 2, wherein the first target-binding domain and the second target-binding domain are Fab molecules.

[0284] 4. The antigen-binding molecule pair according to Embodiment 3, wherein the first target-binding domain comprises a heavy chain variable domain (VH1), a light chain variable domain (VL1), a heavy chain constant domain (CH11), and a light chain constant domain (CL1), and the second target-binding domain comprises a heavy chain variable domain (VH2), a light chain variable domain (VL2), a heavy chain constant domain (CH12), and a light chain constant domain (CL2).

[0285] 5. The antigen-binding molecule pair according to Embodiment 3 or 4, wherein the first target-binding domain and / or the second target-binding domain is a cross-Fab molecule.

[0286] 6. An antigen-binding molecule pair according to any one of Embodiments 1 to 5, wherein the first target-binding domain and the second target-binding domain specifically bind to a tumor-associated antigen or a T cell antigen.

[0287] 7. An antigen-binding molecule pair according to any one of Embodiments 1 to 6, wherein the first target-binding domain and the second target-binding domain specifically bind to FAP, PD-1, Her2, Her3, LAG-3, or EGFR.

[0288] 8.a) The first target-binding domain includes VH1 of SEQ ID NO: 20 and VL1 of SEQ ID NO: 21, and the second target-binding domain includes VH2 of SEQ ID NO: 22 and VL2 of SEQ ID NO: 23, or b) The first target-binding domain includes VH1 of SEQ ID NO: 22 and VL1 of SEQ ID NO: 23, and the second target-binding domain includes VH2 of SEQ ID NO: 20 and VL2 of SEQ ID NO: 21, or c) The first target-binding domain includes VH1 of SEQ ID NO: 76 and VL1 of SEQ ID NO: 77, and the second target-binding domain includes VH2 of SEQ ID NO: 78 and sequence number d) The first target-binding domain includes VH1 of sequence number 78 and VL1 of sequence number 79, and the second target-binding domain includes VH2 of sequence number 76 and VL2 of sequence number 77, or e) The first target-binding domain includes VH1 of sequence number 93 and VL1 of sequence number 94, and the second target-binding domain includes VH2 of sequence number 78 and VL2 of sequence number 79, or f) The first target-binding domain includes VH1 of sequence number 78 and VL1 of sequence number 79, and the second target-binding domain includes sequence number g) The first target-binding domain includes VH2 of SEQ ID NO. 93 and VL2 of SEQ ID NO. 94, or the second target-binding domain includes VH1 of SEQ ID NO. 132 and VL2 of SEQ ID NO. 133, or the first target-binding domain includes VH1 of SEQ ID NO. 132 and VL1 of SEQ ID NO. 133, and the second target-binding domain includes VH2 of SEQ ID NO. 130 and VL2 of SEQ ID NO. 131, or the first target-binding domain includes VH1 of SEQ ID NO. 140 and VL 1) The first target-binding domain includes VH2 of SEQ ID NO: 142 and VL2 of SEQ ID NO: 143, or (j) The first target-binding domain includes VH1 of SEQ ID NO: 142 and VL1 of SEQ ID NO: 143, and the second target-binding domain includes VH2 of SEQ ID NO: 140 and VL2 of SEQ ID NO: 141, or (k) The first target-binding domain includes VH1 of SEQ ID NO: 114 and VL1 of SEQ ID NO: 115, and the second target-binding domain includes VH2 of SEQ ID NO: 116 and VL2 of SEQ ID NO: 117, or (l) The first target-binding domain includes,An antigen-binding molecule pair according to any one of Embodiments 1 to 7, comprising VH1 of SEQ ID NO: 116 and VL1 of SEQ ID NO: 117, wherein the second target-binding domain comprises VH2 of SEQ ID NO: 114 and VL2 of SEQ ID NO: 115, or (m) the first target-binding domain comprises VH1 of SEQ ID NO: 150 and VL1 of SEQ ID NO: 151, wherein the second target-binding domain comprises VH2 of SEQ ID NO: 152 and VL2 of SEQ ID NO: 153, or (n) the first target-binding domain comprises VH1 of SEQ ID NO: 152 and VL1 of SEQ ID NO: 153, wherein the second target-binding domain comprises VH2 of SEQ ID NO: 150 and VL2 of SEQ ID NO: 151.

[0289] 9. An antigen-binding molecule pair according to any one of embodiments 1 to 8, wherein both the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of an IFNγ receptor complex, an IL-2 receptor complex, or an IL-7 receptor complex.

[0290] 10. An antigen-binding molecule pair according to any one of Embodiments 1 to 9, wherein a) the first cytokine receptor-binding domain can bind to IFNγR1 and the second cytokine receptor-binding domain can bind to IFNγR2, or b) the first cytokine receptor-binding domain can bind to IFNγR2 and the second cytokine receptor-binding domain can bind to IFNγR1; or c) the first cytokine receptor-binding domain can bind to IL-2Rβ and the second cytokine receptor-binding domain can bind to IL-2Rγ, or d) the first cytokine receptor-binding domain can bind to IL-2Rγ and the second cytokine receptor-binding domain can bind to IL-2Rβ; or e) the first cytokine receptor-binding domain can bind to IL-2Rγ and the second cytokine receptor-binding domain can bind to IL-7Rα; or f) the first cytokine receptor-binding domain can bind to IL-7Rα and the second cytokine receptor-binding domain can bind to IL-2Rγ.

[0291] 11. An antigen-binding molecule pair according to any one of Embodiments 1 to 10, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are antibody fragments such as Fv, Fab, scFv, scFab, or a single-domain antibody.

[0292] 12. An antigen-binding molecule pair according to any one of Embodiments 1 to 11, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are VHH domains.

[0293] 13. a) The first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, or b) The first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5, or c) The first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, and SEQ ID NO: 60, and the second cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, or d) The first cytokine receptor binding domain includes an amino acid sequence selected from SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, and the second cytokine receptor binding domain The first cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 56, SEQ ID NOs. 57, SEQ ID NOs. 58, SEQ ID NOs. 60, or (e) the first cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 177, SEQ ID NOs. 178, SEQ ID NOs. 179, SEQ ID NOs. 180, SEQ ID NOs. 181, SEQ ID NOs. 182, SEQ ID NOs. 183, SEQ ID NOs. 184, SEQ ID NOs. 185, SEQ ID NOs. 186, SEQ ID NOs. 187, SEQ ID NOs. 188, SEQ ID NOs. 189, SEQ ID NOs. 190, SEQ ID NOs. 191, SEQ ID NOs. 192, and the second cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 61, SEQ ID NOs. f) The first cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 62, SEQ ID NOs. 63, SEQ ID NOs. 64, SEQ ID NOs. 65, and the second cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 61, SEQ ID NOs. 62, SEQ ID NOs. 63, SEQ ID NOs. 64, SEQ ID NOs. 65, and the second cytokine receptor binding domain contains an amino acid sequence selected from SEQ ID NOs. 177, SEQ ID NOs. 178, SEQ ID NOs. 179, SEQ ID NOs. 180, SEQ ID NOs. 181, SEQ ID NOs. 182, SEQ ID NOs. 183, SEQ ID NOs. 184, SEQ ID NOs. 185, SEQ ID NOs. 186, SEQ ID NOs. 187, SEQ ID NOs. 188, SEQ ID NOs. 189, SEQ ID NOs. 190,An antigen-binding molecule pair according to any one of Embodiments 1 to 12, comprising an amino acid sequence selected from SEQ ID NO: 191 and SEQ ID NO: 192.

[0294] 14. an antigen-binding molecule pair according to any one of Embodiments 1 to 13, wherein the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 3 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 7, or b) the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 7 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 3, or c) the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 60 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 62, or d) the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 62 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 60, or e) the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 186 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 62, or f) the first cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 62 and the second cytokine receptor-binding domain comprises the sequence of SEQ ID NO: 186.

[0295] 15. An antigen-binding molecule pair according to any one of Embodiments 1 to 14, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule comprise a first Fc domain subunit and a second Fc domain subunit.

[0296] 16. An antigen-binding molecule pair according to any one of Embodiments 1 to 15, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are Fc domains of IgG, particularly IgG1.

[0297] 17. An antigen-binding molecule pair according to any one of Embodiments 1 to 16, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are human Fc domains.

[0298] 18. An antigen-binding molecule pair according to any one of Embodiments 1 to 17, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include modifications that facilitate association between the first subunit and the second subunit of the Fc domain.

[0299] 19. An antigen-binding molecule pair according to any one of Embodiments 1 to 18, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include one or more amino acid substitutions that reduce binding to and / or effector function of the Fc receptor.

[0300] 20. An antigen-binding molecule pair according to any one of Embodiments 1 to 19, wherein cytokine receptor-binding domains are fused to their respective fusion sites via peptide linkers.

[0301] 21. The antigen-binding molecule pair according to Embodiment 20, wherein the peptide linker comprises an amino acid sequence selected from SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 279, SEQ ID NO: 280, or SEQ ID NO: 281.

[0302] 22. An antigen-binding molecule pair according to any one of Embodiments 1 to 21, wherein a first cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of VH1 or VL1 of the first target-binding domain, and a second cytokine receptor-binding domain is fused at its C-terminus to the N-terminus of VH2 or VL2 of the second target-binding domain.

[0303] 23.a) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor-binding domain, VH1, CH11 and a second Fc domain subunit, in order from the N-terminus to the C-terminus, and a third polypeptide containing VL1 and CL1, in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second cytokine receptor-binding domain, VH2, CH12 and a) The first antigen-binding molecule comprises a second polypeptide containing a first Fc domain subunit, a third polypeptide containing VL2 and CL2 in order from the N-terminus to the C-terminus, or a) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a first cytokine receptor-binding domain, VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises the first Fc domain The first polypeptide comprises a first polypeptide containing a main subunit, a second polypeptide containing VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a second cytokine receptor binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus, or c) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and a first cytokine receptor binding domain, VH1, CH11 and a second Fc domain subunit in order from the N-terminus to the C-terminus. The second antigen-binding molecule comprises a second polypeptide and a third polypeptide containing VL1 and CL1 in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH2, CH12 and a second Fc domain subunit in order from the N-terminus to the C-terminus, and a third polypeptide containing a second cytokine receptor-binding domain, VL2 and CL2 in order from the N-terminus to the C-terminus, where VH1, VL1, CH11 and CL1 form a first target-binding domain, and VH2,VL2, CH12, and CL2 form a second target-binding domain in the antigen-binding molecule pair according to any one of embodiments 1 to 22.

[0304] 24.a) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing a first cytokine receptor-binding domain, VH1, CH11 and a second Fc domain subunit, in order from the N-terminus to the C-terminus, and a third polypeptide containing VL1 and CL1, in order from the N-terminus to the C-terminus, and the second antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second cytokine receptor-binding domain, VH2, CH1 a) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11, and a second Fc domain subunit, and a third polypeptide containing VL2 and CL2 in order from the N-terminus to the C-terminus, or a) the first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, a second polypeptide containing VH1, CH11, and a second Fc domain subunit, in order from the N-terminus to the C-terminus, and a third polypeptide containing a first...

Claims

1. A pair of antigen-binding molecules that specifically bind to a target antigen, a) i) a first antigen-binding molecule comprising a first target-binding domain, ii) a first cytokine receptor-binding domain, and iii) an Fc domain, and b) comprising a second antigen-binding molecule comprising i) a second target-binding domain, ii) a second cytokine receptor-binding domain, and iii) an Fc domain, The first target-binding domain can bind to a first epitope on the target antigen, the second target-binding domain can bind to a second epitope on the target antigen, the first target-binding domain and the second target-binding domain do not compete for binding to the target antigen, and The first cytokine receptor binding domain can bind to a first cytokine receptor subunit, and the second cytokine receptor binding domain can bind to a second cytokine receptor subunit. Antigen-binding molecule pair.

2. The antigen-binding molecule pair according to claim 1, wherein the first target-binding domain and the second target-binding domain are antibody fragments such as Fv, Fab, scFv, scFab molecules, or single-domain antibodies.

3. The antigen-binding molecule pair according to claim 1 or 2, wherein the first target-binding domain and the second target-binding domain are Fab molecules.

4. The first target-binding domain is a heavy chain variable domain (VH １ ), light chain variable domain (VL １ ), heavy chain constant domain (CH1 １ ) and light chain constant domain (CL １ ) includes, and the second target binding domain is a heavy chain variable domain (VH ２ ), light chain variable domain (VL ２ ), heavy chain constant domain (CH1 ２ ) and light chain constant domain (CL ２ The antigen-binding molecule pair according to claim 3, comprising )

5. The antigen-binding molecule pair according to claim 3 or 4, wherein the first target-binding domain and / or the second target-binding domain is a cross-Fab molecule.

6. The antigen-binding molecule pair according to any one of claims 1 to 5, wherein the first target-binding domain and the second target-binding domain specifically bind to a tumor-associated antigen or a T cell antigen.

7. The antigen-binding molecule pair according to any one of claims 1 to 6, wherein the first target-binding domain and the second target-binding domain specifically bind to FAP, PD-1, Her2, Her3, LAG-3, or EGFR.

8. An antigen-binding molecule pair according to any one of claims 1 to 7, a) the first target binding domain comprises VH of SEQ ID NO: 20 １ and VL of SEQ ID NO: 21 １ and the second target binding domain comprises VH of SEQ ID NO: 22 ２ and VL of SEQ ID NO: 23 ２ or b) The first target-binding domain is the VH of Sequence ID No.

22. １ and VL of Sequence ID No. 23 １ The second target-binding domain is VH of SEQ ID NO: 20 ２ and VL of sequence number 21 ２ Includes, or c) The first target-binding domain is the VH of Sequence ID No.

76. １ and VL of Sequence ID 77 １ The second target-binding domain is VH of Sequence ID No.

78. ２ and VL of Sequence ID 79 ２ Includes, or d) The first target-binding domain is the VH of Sequence ID No.

78. １ and VL of Sequence ID 79 １ The second target-binding domain is VH of Sequence ID No.

76. ２ and VL of Sequence ID 77 ２ Includes, or e) The first target-binding domain is the VH of Sequence ID No.

93. １ and VL of sequence number 94 １ The second target-binding domain is VH of Sequence ID No.

78. ２ and VL of Sequence ID 79 ２ Includes, or f) The first target-binding domain is the VH of Sequence ID No.

78. １ and VL of Sequence ID 79 １ The second target-binding domain is VH of Sequence ID No.

93. ２ and VL of sequence number 94 ２ Includes, or g) The first target-binding domain is VH of SEQ ID NO: 130 １ and VL of sequence number 131 １ The second target-binding domain is VH of SEQ ID NO:

132. ２ and VL of sequence number 133 ２ Includes, or h) The first target-binding domain is VH of SEQ ID NO: 132 １ and VL of sequence number 133 １ The second target-binding domain is VH of SEQ ID NO:

130. ２ and VL of sequence number 131 ２ Includes, or i) The first target-binding domain is VH of Sequence ID No.

140. １ and VL of sequence number 141 １ The second target-binding domain is VH of SEQ ID NO:

142. ２ and VL of sequence number 143 ２ Includes, or j) The first target-binding domain is VH of SEQ ID NO: 142 １ and VL of sequence number 143 １ The second target-binding domain is VH of SEQ ID NO:

140. ２ and VL of sequence number 141 ２ Includes, or k) The first target-binding domain is VH of SEQ ID NO: 114 １ and VL of sequence number 115 １ The second target-binding domain is VH of SEQ ID NO:

116. ２ and VL of sequence number 117 ２ Includes, or l) The first target-binding domain is VH of SEQ ID NO: 116 １ and VL of sequence number 117 １ The second target-binding domain is VH of SEQ ID NO:

114. ２ and VL of sequence number 115 ２ Includes, or m) The first target-binding domain is VH of Sequence ID No.

150. １ and VL of sequence number 151 １ The second target-binding domain is VH of SEQ ID NO:

152. ２ and VL of sequence number 153 ２ Includes, or n) The first target-binding domain is VH of SEQ ID NO: 152 １ and VL of sequence number 153 １ The second target-binding domain is VH of SEQ ID NO:

150. ２ and VL of sequence number 151 ２ including, Antigen-binding molecule pair.

9. The antigen-binding molecule pair according to any one of claims 1 to 8, wherein both the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of an IFNγ receptor complex, an IL-2 receptor complex, or an IL-7 receptor complex.

10. An antigen-binding molecule pair according to any one of claims 1 to 9, a) The first cytokine receptor binding domain can bind to IFNγR1, and the second cytokine receptor binding domain can bind to IFNγR2, or b) The first cytokine receptor binding domain can bind to IFNγR2 and the second cytokine receptor binding domain can bind to IFNγR1, or c) The first cytokine receptor binding domain can bind to IL-2Rβ, and the second cytokine receptor binding domain can bind to IL-2Rγ, or d) The first cytokine receptor binding domain can bind to IL-2Rγ, and the second cytokine receptor binding domain can bind to IL-2Rβ, or e) The first cytokine receptor binding domain can bind to IL-2Rγ, and the second cytokine receptor binding domain can bind to IL-7Rα, or f) The first cytokine receptor binding domain can bind to IL-7Rα, and the second cytokine receptor binding domain can bind to IL-2Rγ, Antigen-binding molecule pair.

11. The antigen-binding molecule pair according to any one of claims 1 to 10, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are antibody fragments such as Fv, Fab, scFv, scFab, or a single-domain antibody.

12. The antigen-binding molecule pair according to any one of claims 1 to 11, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are VHH domains.

13. An antigen-binding molecule pair according to any one of claims 1 to 12, a) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 5, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10, or b) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 5, or c) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, and SEQ ID NO: 60, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, or d) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, and SEQ ID NO: 60, or e) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NOs: 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, and 192, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NOs: 61, 62, 63, 64, and 65, or f) The first cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NOs: 61, 62, 63, 64, and 65, and the second cytokine receptor binding domain comprises an amino acid sequence selected from SEQ ID NOs: 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, and 192. Antigen-binding molecule pair.

14. An antigen-binding molecule pair according to any one of claims 1 to 13, a) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 3, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 7, or b) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 7, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 3, or c) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 60, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, or d) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 60, or e) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 186, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, or f) The first cytokine receptor binding domain includes the sequence of SEQ ID NO: 62, and the second cytokine receptor binding domain includes the sequence of SEQ ID NO: 186, Antigen-binding molecule pair.

15. The antigen-binding molecule pair according to any one of claims 1 to 14, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule comprise a first Fc domain subunit and a second Fc domain subunit.

16. The Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are IgG, particularly IgG １ An antigen-binding molecule pair according to any one of claims 1 to 15, wherein the Fc domain is...

17. The antigen-binding molecule pair according to any one of claims 1 to 16, wherein the Fc domain of the first antigen-binding molecule and the second antigen-binding molecule is a human Fc domain.

18. The antigen-binding molecule pair according to any one of claims 1 to 17, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include modifications that promote association between the first subunit and the second subunit of the Fc domain.

19. The antigen-binding molecule pair according to any one of claims 1 to 18, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include one or more amino acid substitutions that reduce binding to and / or effector function of the Fc receptor.

20. The antigen-binding molecule pair according to any one of claims 1 to 19, wherein the cytokine receptor-binding domain is fused to each fusion site via a peptide linker.

21. The antigen-binding molecule pair according to claim 20, wherein the peptide linker comprises an amino acid sequence selected from SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQ ID NO: 279, SEQ ID NO: 280, or SEQ ID NO:

281.

22. The first cytokine receptor binding domain has a VH at its C-terminus, the same as the first target binding domain. １ or VL １ It is fused to the N-terminus, and the second cytokine receptor binding domain is at its C-terminus, the VH of the second target binding domain. ２ or VL ２ A pair of antigen-binding molecules according to any one of claims 1 to 21, fused to the N-terminus of the following.

23. An antigen-binding molecule pair according to any one of claims 1 to 22, a) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VH １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor-binding domain, VH ２ CH1 ２ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. ２ and CL ２ A third polypeptide comprising, or b) The first antigen-binding molecule has a first polypeptide containing a first Fc domain subunit, and VH １ CH1 １ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the first cytokine receptor binding domain, VL １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor binding domain, VL ２ and CL ２ A third polypeptide comprising, or c) The first antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, and, in order from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VH １ , CH1 １ and a second polypeptide comprising a second Fc domain subunit, and, in order from the N-terminus to the C-terminus, VL １ and CL １ and a third polypeptide comprising the third polypeptide, and the second antigen-binding molecule comprises a first polypeptide comprising a first Fc domain subunit, and, in order from the N-terminus to the C-terminus, VH ２ , CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and, in order from the N-terminus to the C-terminus, the second cytokine receptor-binding domain, VL ２ and CL ２ and a third polypeptide comprising the third polypeptide, VH １ 、VL １ 、CH1 １ 以及CL １ 形成所述第一靶结合结构域,VH ２ 、VL ２ 、CH1 ２ 以及CL ２ 形成所述第二靶结合结构域。 Antigen-binding molecule pair.

24. An antigen-binding molecule pair according to any one of claims 1 to 23, a) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VH １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor-binding domain, VH ２ CH1 ２ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. ２ and CL ２ A third polypeptide comprising, or b) The first antigen-binding molecule has a first polypeptide containing a first Fc domain subunit, and VH １ CH1 １ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the first cytokine receptor binding domain, VL １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor binding domain, VL ２ and CL ２ A third polypeptide comprising, or c) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and, sequentially from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VH １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor binding domain, VL ２ and CL ２ A third polypeptide comprising, or d) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and, sequentially from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VL １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor-binding domain, VH ２ CH1 ２ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. ２ and CL ２ A third polypeptide comprising, or e) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ CH1 １ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the first cytokine receptor binding domain, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor binding domain, VL ２ and CL ２ A third polypeptide comprising, or f) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and, sequentially from the N-terminus to the C-terminus, the first cytokine receptor-binding domain, VL １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a second polypeptide comprising a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, the second cytokine receptor binding domain, VL ２ and CL ２ A third polypeptide comprising, or g) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ CH1 １ and a second polypeptide containing a second Fc domain subunit, and sequentially from the N-terminus to the C-terminus, a first cytokine receptor binding domain, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, a second cytokine receptor-binding domain, VH ２ CH1 ２ and a second polypeptide containing a second Fc domain subunit, and VL in order from the N-terminus to the C-terminus. ２ and CL ２ A third polypeptide comprising, VH １ , VL １ CH1 １ and CL １ This forms the first target-binding domain, VH ２ , VL ２ CH1 ２ and CL ２ The second target-binding domain is formed by Antigen-binding molecule pair.

25. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to FAP, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IFNγ receptor complex.

26. An antigen-binding molecule pair according to any one of claims 1 to 25, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 27, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 30, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 38, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 35, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 27, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 33, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 30, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 31, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 38, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 36, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 37, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

35. Antigen-binding molecule pair.

27. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to EGFR, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IFNγ receptor complex.

28. An antigen-binding molecule pair according to any one of claims 1 to 24 or 27, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 260, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 256, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 122, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 255, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 257, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 254, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 260, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 255, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 256, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 259, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 127, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 261, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 254, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 119, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 257, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 258, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

127. Antigen-binding molecule pair.

29. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to PD-1, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

30. An antigen-binding molecule pair according to any one of claims 1 to 24 or 29, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 86, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 83, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 87, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 92, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 91, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 95, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 99, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 100, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 104, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 98, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 99, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 101, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 95, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 102, or i) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 100, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 98, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 103, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or j) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 96, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 97, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 88, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

104. Antigen-binding molecule pair.

31. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to LAG-3, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

32. An antigen-binding molecule pair according to any one of claims 1 to 24 or 31, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

157. Antigen-binding molecule pair.

33. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to EGFR, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

34. An antigen-binding molecule pair according to any one of claims 1 to 24 or 33, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 157, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 161, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 165, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 154, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 159, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 162, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 160, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 164, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 163, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 155, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 156, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 158, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

157. Antigen-binding molecule pair.

35. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to FAP, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

36. An antigen-binding molecule pair according to any one of claims 1 to 24 or 35, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 111, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 110, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 109, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 105, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 113, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 111, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 108, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 107, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 112, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

29. Antigen-binding molecule pair.

37. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to Her2, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

38. An antigen-binding molecule pair according to any one of claims 1 to 24 or 37, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 136, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 134, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 138, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 137, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 135, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 134, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 138, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

139. Antigen-binding molecule pair.

39. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to Her3, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-2 receptor complex.

40. An antigen-binding molecule pair according to any one of claims 1 to 24 or 39, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 146, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 144, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 148, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 147, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 145, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 144, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 148, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

149. Antigen-binding molecule pair.

41. The antigen-binding molecule pair according to any one of claims 1 to 24, wherein the first target-binding domain and the second target-binding domain specifically bind to PD-1, and the first cytokine receptor subunit and the second cytokine receptor subunit are subunits of the IL-7 receptor complex.

42. An antigen-binding molecule pair according to any one of claims 1 to 24 and 41, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 194, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 193, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 195, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 196, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 197, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 198, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 199, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 200, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 201, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 202, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or i) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 203, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or j) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 204, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 89, or k) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 106, a second polypeptide containing the amino acid sequence of SEQ ID NO: 205, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 124, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 90, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

89. Antigen-binding molecule pair.

43. The first target-binding domain is CH1 １ The first cytokine receptor binding domain is fused at its C-terminus to the N-terminus of the first Fc domain subunit, and the second target binding domain is CH1 ２ The antigen-binding molecule pair according to any one of claims 1 to 21, wherein the C-terminus of the first Fc domain subunit is fused to the N-terminus of the second Fc domain subunit, and the second cytokine receptor-binding domain is fused to the N-terminus of the second Fc domain subunit at its C-terminus.

44. An antigen-binding molecule pair according to any one of claims 1 to 21 or 43, a) The first antigen-binding molecule is VH １ CH1 １ and a first polypeptide including a first Fc domain subunit, and VL sequentially from the N-terminus to the C-terminus. １ and CL １ The second polypeptide comprises a second polypeptide containing and a third polypeptide containing the first cytokine receptor binding domain and the second Fc domain subunit in order from the N-terminus to the C-terminus, wherein the second antigen-binding molecule comprises VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a first polypeptide including a first Fc domain subunit, and VL sequentially from the N-terminus to the C-terminus. ２ and CL ２ A second polypeptide comprising a second polypeptide containing a second cytokine receptor-binding domain and a third polypeptide comprising a second cytokine receptor-binding domain and a second Fc domain subunit in order from the N-terminus to the C-terminus, or b) The first antigen-binding molecule is VL in order from the N-terminus to the C-terminus. １ CH1 １ and a first polypeptide comprising a first Fc domain subunit, and sequentially from the N-terminus to the C-terminus, VH １ and CL １ The second polypeptide comprises a second polypeptide containing and a third polypeptide containing the first cytokine receptor binding domain and the second Fc domain subunit in order from the N-terminus to the C-terminus, wherein the second antigen-binding molecule comprises VH in order from the N-terminus to the C-terminus. ２ CH1 ２ and a first polypeptide including a first Fc domain subunit, and VL sequentially from the N-terminus to the C-terminus. ２ and CL ２ The second polypeptide comprises a second polypeptide containing a second cytokine receptor binding domain and a third polypeptide comprising a second cytokine receptor binding domain and a second Fc domain subunit, in order from the N-terminus to the C-terminus. VH １ , VL １ CH1 １ and CL １ This forms the first target-binding domain, VH ２ , VL ２ CH1 ２ and CL ２ The second target-binding domain is formed by Antigen-binding molecule pair.

45. An antigen-binding molecule pair according to any one of claims 1 to 21 or 43 to 44, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 32, a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 39, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 34, a second polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 34, a second polypeptide containing the amino acid sequence of SEQ ID NO: 29, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 39, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 32, a second polypeptide containing the amino acid sequence of SEQ ID NO: 25, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 40, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 238, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 236, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 239, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 240, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 237, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 236, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 235, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 240, a second polypeptide containing the amino acid sequence of SEQ ID NO: 32, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 239, a second polypeptide containing the amino acid sequence of SEQ ID NO: 34, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

29. Antigen-binding molecule pair.

46. The first target-binding domain is CH1 １ The first cytokine receptor binding domain is fused at its C-terminus to the N-terminus of one of the Fc domain subunits, and the second target binding domain is CH1 ２ The antigen-binding molecule pair according to any one of claims 1 to 21, wherein the C-terminus of one of the Fc domain subunits is fused to the N-terminus of the second cytokine receptor-binding domain, and the N-terminus of the second cytokine receptor-binding domain is fused to the C-terminus of the same Fc domain subunit.

47. An antigen-binding molecule pair according to any one of claims 1 to 21 or 46, a) The first antigen-binding molecule has a first polypeptide containing a first Fc domain subunit, and VH １ CH1 １ , a second polypeptide comprising a second Fc domain subunit and the first cytokine receptor binding domain, and VL in order from the N-terminus to the C-terminus. １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ , a second polypeptide comprising a second Fc domain subunit and the second cytokine receptor binding domain, and sequentially from the N-terminus to the C-terminus, VL ２ and CL ２ A third polypeptide comprising, or b) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ CH1 １ , a second polypeptide comprising a second Fc domain subunit and the first cytokine receptor binding domain, and sequentially from the N-terminus to the C-terminus, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ , a second polypeptide comprising a second Fc domain subunit and the second cytokine receptor binding domain, and sequentially from the N-terminus to the C-terminus, VL ２ and CL ２ A third polypeptide comprising, or c) The first antigen-binding molecule comprises a first polypeptide containing a first Fc domain subunit, and VL in order from the N-terminus to the C-terminus. １ CH1 １ , a second polypeptide comprising a second Fc domain subunit and the first cytokine receptor binding domain, and sequentially from the N-terminus to the C-terminus, VH １ and CL １ The second antigen-binding molecule comprises a third polypeptide containing a first Fc domain subunit, and VH in order from the N-terminus to the C-terminus. ２ CH1 ２ , a second polypeptide comprising a second Fc domain subunit and the second cytokine receptor binding domain, and sequentially from the N-terminus to the C-terminus, VL ２ and CL ２ A third polypeptide comprising, VH １ , VL １ CH1 １ and CL １ This forms the first target-binding domain, VH ２ , VL ２ CH1 ２ and CL ２ The second target-binding domain is formed by Antigen-binding molecule pair.

48. An antigen-binding molecule pair according to any one of claims 1 to 21 or 46 to 47, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 41, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 45, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 43, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 46, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 41, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 47, a second polypeptide containing the amino acid sequence of SEQ ID NO: 44, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 43, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 246, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 249, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 242, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 245, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or e) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 250, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 253, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or f) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 248, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 247, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or g) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 244, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 243, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or h) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 252, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 25, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 251, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 29, or i) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 28, a second polypeptide containing the amino acid sequence of SEQ ID NO: 42, and a third polypeptide containing the amino acid sequence of SEQ ID NO: 42, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 241, a second polypeptide containing the amino acid sequence of SEQ ID NO: 245, and a third polypeptide containing the amino acid sequence of SEQ ID NO:

29. Antigen-binding molecule pair.

49. The antigen-binding molecule pair according to any one of claims 1 to 11, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are Fab molecules.

50. The antigen-binding molecule pair according to claim 49, a) The first cytokine receptor binding domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 282 and a VL domain containing the amino acid sequence of SEQ ID NO: 283, or a VH domain containing the amino acid sequence of SEQ ID NO: 284 and a VL domain containing the amino acid sequence of SEQ ID NO: 285, and the second cytokine receptor domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 286 and a VL domain containing the amino acid sequence of SEQ ID NO: 287, or a VH domain containing the amino acid sequence of SEQ ID NO: 288 and a VL domain containing the amino acid sequence of SEQ ID NO: 289, or b) The first cytokine receptor binding domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 286 and a VL domain containing the amino acid sequence of SEQ ID NO: 287, or a VH domain containing the amino acid sequence of SEQ ID NO: 288 and a VL domain containing the amino acid sequence of SEQ ID NO: 289, and the second cytokine receptor domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 282 and a VL domain containing the amino acid sequence of SEQ ID NO: 283, or a VH domain containing the amino acid sequence of SEQ ID NO: 284 and a VL domain containing the amino acid sequence of SEQ ID NO: 285.16, Antigen-binding molecule pair.

51. The antigen-binding molecule pair according to any one of claims 1 to 11, wherein the first cytokine receptor-binding domain and the second cytokine receptor-binding domain are scFv molecules.

52. The antigen-binding molecule pair according to claim 51, a) The first cytokine receptor binding domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 290 and a VL domain containing the amino acid sequence of SEQ ID NO: 291, and the second cytokine receptor domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 292 and a VL domain containing the amino acid sequence of SEQ ID NO: 293, or b) The first cytokine receptor binding domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 292 and a VL domain containing the amino acid sequence of SEQ ID NO: 293, and the second cytokine receptor domain comprises a VH domain containing the amino acid sequence of SEQ ID NO: 290 and a VL domain containing the amino acid sequence of SEQ ID NO:

291. Antigen-binding molecule pair.

53. The antigen-binding molecule pair according to any one of claims 49 to 52, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule comprise a first Fc domain subunit and a second Fc domain subunit.

54. The Fc domains of the first antigen-binding molecule and the second antigen-binding molecule are IgG, particularly IgG １ The antigen-binding molecule pair according to any one of claims 49 to 53, wherein the Fc domain is...

55. The antigen-binding molecule pair according to any one of claims 49 to 54, wherein the Fc domain of the first antigen-binding molecule and the second antigen-binding molecule is a human Fc domain.

56. The antigen-binding molecule pair according to any one of claims 49 to 55, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include modifications that promote association between the first subunit and the second subunit of the Fc domain.

57. The antigen-binding molecule pair according to any one of claims 49 to 56, wherein the Fc domains of the first antigen-binding molecule and the second antigen-binding molecule include one or more amino acid substitutions that reduce binding to and / or effector function of the Fc receptor.

58. The antigen-binding molecule pair according to any one of claims 49 to 57, wherein the cytokine receptor-binding domain is fused to each fusion site via a peptide linker.

59. The antigen-binding molecule pair according to claim 58, wherein the peptide linker comprises the amino acid sequence of SEQ ID NO: 280 or SEQ ID NO:

281.

60. The antigen-binding molecule pair according to claim 49, a) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 267, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 266, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 272, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 271, or b) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 267, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 266, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 274, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 273, or c) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 269, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 268, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 268, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 271, or d) The first antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 269, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 265, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO: 268, and the second antigen-binding molecule comprises a first polypeptide containing the amino acid sequence of SEQ ID NO: 274, a second polypeptide containing the amino acid sequence of SEQ ID NO: 28, a third polypeptide containing the amino acid sequence of SEQ ID NO: 270, and a fourth polypeptide containing the amino acid sequence of SEQ ID NO:

273. Antigen-binding molecule pair.

61. The antigen-binding molecule pair according to claim 51, wherein the first antigen-binding molecule comprises a first polypeptide having the amino acid sequence of SEQ ID NO: 276, a second polypeptide having the amino acid sequence of SEQ ID NO: 28, and a third polypeptide having the amino acid sequence of SEQ ID NO: 275, and the second antigen-binding molecule comprises a first polypeptide having the amino acid sequence of SEQ ID NO: 277, a second polypeptide having the amino acid sequence of SEQ ID NO: 28, and a third polypeptide having the amino acid sequence of SEQ ID NO: 278.

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