RU2824467C2 - Fc-binding fragments containing antigen-binding site for cd137 - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: described is a group of inventions comprising a specifically binding element which binds CD137, a nucleic acid molecule coding a specifically binding element, a recombinant host cell containing a nucleic acid, to obtain a specifically binding element, a method for producing a specifically binding element, comprising culturing a recombinant host cell, use of a specific binding element for treating cancer in an individual and use of a specific binding element for treating an infectious disease in an individual.

EFFECT: invention extends the range of agents specifically binding to CD137.

21 cl, 15 dwg, 17 tbl, 13 ex

Description

Field of technology

The present invention relates to specific binding elements that bind CD137. The specific binding elements comprise an antigen-binding site for CD137 located in the constant domain of the specific binding element and are used, by way of example, in the treatment of cancer and infectious diseases.

State of the art

Cell signaling is an integral part of life in all organisms and typically involves cell surface receptors that interact with soluble or surface-expressed ligands. This interaction results in changes in the receptor, the ligand, or both. For example, ligand binding can induce conformational changes in receptors, causing them to cluster into dimers or oligomers. This clustering effect then leads to the activation of intracellular signaling pathways. Numerous receptors are activated in this way, including members of the tumor necrosis factor receptor superfamily (TNFRSF), such as CD137.

CD137 (4-1BB; TNFRSF9) is a costimulatory molecule of the tumor necrosis factor receptor superfamily (TNFRSF). CD137 is well known to be upregulated on CD8 ⁺ T cells following activation and can also be expressed on activated CD4 ⁺ helper T cells, B cells, regulatory T cells, natural killer (NK) cells, natural killer T cells (NKT cells), and dendritic cells (DCs) (Bartkowiak & Curran, 2015). The essential functional role of CD137 in enhancing T cell cytotoxicity was first described in 1997 (Shuford et al., 1997), and shortly thereafter, CD137 monoclonal antibodies (MAbs) were proposed as anticancer therapeutics.

CD137 is a transmembrane protein with four extracellular cysteine-rich domains termed CRD1-4 and a cytoplasmic region responsible for CD137 signaling. The ligand for CD137 is CD137L. Although no crystal structure exists for the CD137/CD137L complex, CD137 is predicted to form a trimer/trimer complex with CD137L (Won et al., 2010). Binding of CD137L results in the formation of a receptor trimer and subsequent clustering of multiple receptor trimers, and leads to activation of the CD137 signaling cascade. This signaling cascade provides a survival signal to T cells by counteracting activation-induced cell death (Hurtado et al., 1997), thereby playing a critical role in maintaining effective T cell immune responses and establishing immunological memory (Bartkowiak & Curran, 2015).

The role of CD137 in leukocyte biology is generally well understood, with a clear biological rationale for its role in tumor immunology. CD137 is expressed by activated T cells and has been used as a marker to identify antigen-specific CD4 ⁺ and CD8 ⁺ T cells. Generally, CD137 expression is higher on CD8 ⁺ T cells than on CD4 ⁺ T cells (Wen et al., 2002). In the case of CD8 ⁺ T cells, proliferation, survival, and cytotoxic effector function through the production of interferon-gamma and interleukin 2 have been attributed to CD137 clustering. CD137 clustering also contributes to the differentiation and maintenance of memory CD8 ⁺ T cells. In some CD4 ⁺ T cell subsets, CD137 clustering similarly leads to proliferation and activation and results in the release of cytokines such as interleukin 2 (Makkouk et al., 2016).

Natural killer cell (NK) antibody-dependent cellular cytotoxicity (ADCC) mediated by tumor-targeted mAbs has been demonstrated to be enhanced by CD137 stimulation with agonistic anti-CD137 mAbs in vitro and in vivo (Bartkowiak & Curran, 2015). NK cells bind antibodies via their Fc receptor and, depending on the antibody isotype, this can lead to NK cell activation, causing the release of cytotoxic granules and lysis of target cells (Kohrt et al., 2012). Kohrt and colleagues demonstrated that an anti-CD137 agonist antibody enhances the antitumor activity of the therapeutic antibodies rituximab, trastuzumab, and cetuximab by enhancing ADCC when dosed in combination with them (Kohrt et al., 2014; Kohrt et al., 2011). Furthermore, in human NK cells, CD137 expression is upregulated following exposure to cell-bound antibodies through their FcγR. Subsequent stimulation of these NK cells with an anti-CD137 antibody was shown to enhance their ADCC against tumor cells (Chester et al., 2015; Chester et al., 2016).

B lymphocytes also express CD137 upon activation. Binding of CD137 ligand to CD137 enhances B cell proliferation, survival, and cytokine production. CD137 expression is also induced on normal and malignant human B cells following binding of CD40 to its ligand CD154 (CD40 ligand), resulting in increased B cell survival if CD137 is subsequently activated (Vinay and Kwon, 2011).

CD137 has also been demonstrated to be expressed on tumor-infiltrating lymphocyte (TIL) subsets reactive against tumors. CD137 monotherapy has been shown to be effective in several preclinical models of immunogenic tumors such as MC38, CT26, and B-cell lymphomas. Combination targeting of CD137 with other anti-cancer agents such as chemotherapy, cytokines, and other checkpoint regulators has been shown to result in enhanced growth reduction of established tumors. Specifically, combination of anti-CD137 antibodies with anti-CD20 antibodies, anti-EGFR antibodies, and anti-HER-2 antibodies has been shown to result in synergistic tumor reduction in various preclinical xenograft models (Kohrt et al., 2014; Kohrt et al., 2012; Kohrt et al., 2011).

Combining tumor-targeted monoclonal antibodies with anti-CD137 agonist antibodies has shown promising results in preclinical models of lymphoma (Kohrt et al., 2011), head and neck cancer, colorectal cancer (Kohrt et al., 2014), and breast cancer (Kohrt et al., 2012). A number of tumor-targeted monoclonal antibodies have also been tested in combination with anti-CD137 agonist antibodies in the clinical setting, including the anti-CD20 mAb rituximab (NCT01307267, NCT02951156), the anti-EGFR mAb cetuximab (NCT02110082), and the anti-CS1 mAb elotuzumab (NCT02252263). However, clinical development has been slowed by dose-limiting severe liver inflammation associated with CD137 agonist antibody treatment. Urelumab (BMS-663513), a non-ligand-blocking human IgG4 antibody (Chester et al., 2018), was the first CD137 antibody to enter clinical trials, but these were stopped after significant, as expected, dose-dependent liver toxicity was observed (Chester et al., 2018). More recently, clinical trials of urelumab have been restarted in various solid cancers, combining urelumab with radiotherapy (NCT03431948) or with other therapeutic antibodies, such as rituximab (NCT01775631), cetuximab (NCT02110082), the anti-PD-1 antibody nivolumab (NCT02253992, NCT02534506, NCT02845323), and the combination of nivolumab and the anti-LAG-3 antibody BMS986016 (NCT02658981). However, to reduce urelumab-associated liver toxicity, urelumab dosing had to be limited in these studies, and efficacy results were disappointing (Chester et al., 2018).

No dose-limiting toxicity was observed with Pfizer's anti-CD137 antibody utomilumab (PF-05082566), a human IgG2 antibody, across the dose range of 0.03 mg/kg to 10 mg/kg in phase I clinical trials of advanced cancer (Chester et al. 2016; Segal et al., 2018). However, the overall objective response rate with this antibody was only 3.8% in patients with solid tumors, possibly indicating that utomilumab has weaker activity and clinical efficacy than urelumab but has a more favorable safety profile (Chester et al., 2018; Segal et al., 2018). Utomilumab has been tested in combination with radiation therapy (NCT03217747) or chemotherapy, and in combination with other antibody therapies, including the anti-PD-L1 antibody avelumab (NCT02554812) and the anti-PD-1 antibody pembrolizumab (NCT02179918), to evaluate the safety, tolerability, dose-limiting toxicities (DLTs), maximum tolerated dose (MTD), and efficacy of different treatment combinations. These studies are ongoing, with early results showing no DLTs at doses up to 5 mg/kg and a 26% response rate for the combination of utomilumab and pembrolizumab. Triple combinations of utomilumab with avelumab and other immuno-oncology therapies are also being tested (NCT02554812, NCT03217747).

A number of bispecific molecules targeting CD137 are also in early development, many of which are generated using non-antibody scaffold technology or protein fusion. The development of a bispecific molecule targeting CD137 and FAPalpha using DARPin scaffold protein technology has been reported (Link et al., 2018; Reichen et al., 2018). T cell activation by tumor targeting of CD137 agonist activity has also been demonstrated using HER2- and EphA2-targeting DART molecules (Liu et al., 2017). CD137L fusion proteins that target tumors via FAPalpha or CD19 in solid tumors and lymphomas are also under development. The most clinically advanced CD137 bispecific molecule (and the only one containing a full-length antibody) is PRS-343, a CD137/HER2 bispecific molecule. In this molecule, CD137 is bound by an artificial binding protein (anticalin) attached to the Fc portion of the HER2-targeting IgG4 antibody trastuzumab. In a humanized mouse model, PRS-343 was reported to mediate tumor target-dependent activation of CD137 on lymphocytes at sites where HER2 is overexpressed, but no improvement in tumor growth inhibition was observed compared to trastuzumab alone (Hinner et al., 2016 and WO 2016/177802 A1). PRS-343 recently entered phase I clinical trials in a range of solid tumors to evaluate its safety, tolerability, and efficacy (NCT03330561).

Statement of the essence of the invention

As explained in the background section above, clinical development of CD137 agonist molecules has been stalled due to treatment being either associated with dose-limiting severe liver inflammation (urelumab) or low clinical efficacy (utomilumab).

The inventors of the present invention have determined that there is a need in the art for CD137 agonist molecules that exhibit high activity but are not associated with dose-limiting liver inflammation. Such molecules can be administered to individuals at doses that optimize the activity and therefore the effectiveness of the molecule and can be used in the treatment of cancer, for example, as immunotherapeutic agents, or in the treatment of infectious diseases.

Without being bound by theory, it is believed that T cells present in the liver can potentially be activated by CD137 agonist molecules, leading to liver inflammation. CD8 ⁺ T cells have been shown to promote inflammation and apoptosis in the liver following sepsis/viral infection (Wesche-Soldato et al., 2007). CD137 agonist antibody therapy in mice has been shown to result in CD137-dependent T cell infiltration into the liver (Dubrot J et al., 2010). Collectively, these studies suggest that highly potent CD137 agonist antibodies such as urelumab can induce activated CD8 ⁺ T cell infiltration into the liver, leading to liver inflammation. Alternatively, the dose-limiting liver toxicity observed with urelumab treatment may be due to a specific epitope bound by this antibody.

The present inventors performed an extensive selection and affinity maturation program to isolate a panel of antigen-binding Fc fragments (also referred to herein as “Fcab”) that contain a binding site for CD137 in their CH3 domain and bind dimeric CD137 with higher affinity than monomeric CD137.

In the present application, "affinity" may refer to the strength of the binding interaction between an antibody molecule and its cognate antigen, measured as _KD . As one of skill in the art will readily appreciate, if an antibody molecule is capable of forming multiple binding interactions with an antigen (e.g., if the antibody molecule is capable of bivalently binding an antigen, and optionally, the antigen is dimeric), the affinity, measured as _KD , may also be affected by avidity, wherein avidity refers to the overall strength of the antibody-antigen complex.

Expression of CD137 by T cells is up-regulated upon activation. Without being bound by theory, it is believed that due to the high expression of CD137 on activated T cells, CD137 will be present in the form of dimers, trimers, and higher-order multimers on the surface of such cells. In contrast, naive immune cells, such as naive T cells, express low or negligible levels of CD137 on their cell surface and therefore any CD137 present is likely to be in the monomeric form. Thus, Fcabs that bind to dimeric CD137 with higher affinity than to monomeric CD137 are expected to preferentially bind to activated immune cells, such as activated T cells, as opposed to naive immune cells, as an example.

The Fcabs of the present invention were also able to bind dimeric cynomolgus monkey CD137. This is advantageous because it allows toxicity and efficacy testing to be performed in cynomolgus monkeys during preclinical development. This is a particular advantage for antibody molecules that bind to CD137, given the liver inflammation observed with some CD137 antibodies. Two of the isolated Fcabs, FS22-053-014 and FS22-053-017, also bound mouse CD137. This is advantageous because it allows the same Fcab to be tested in mice before administration to cynomolgus monkeys or humans. Normally, an Fcab that binds to mouse CD137 is needed for this purpose.

The present inventors unexpectedly found that all isolated anti-CD137 Fcabs that preferentially bound dimeric over monomeric CD137 and were capable of affinity maturation contained a PPY motif as well as a 5 amino acid insertion in the AB loop of their CH3 domain. Another anti-CD137 Fcab line isolated after initial library screening did not possess these features and was not capable of affinity maturation and, accordingly, was not used further. Without wishing to be bound by theory, it is believed that the presence of the PPY motif may promote the formation of an extended antigen-binding region by creating a more rigid or open loop structure as a result of the limited flexibility of proline residues. Alternatively, the PPY sequence may represent a specific conserved motif involved in binding to CD137, as proline-rich sequences have been demonstrated to bind aromatic sequences in SH3 domain proteins, as an example. Furthermore, since the conserved PPY sequence was selected independently in two separate Fcab lines, it may be important for CD137 epitope binding. Furthermore, the conserved LE or LD sequence was present in the EF loops of the CH3 domain of most isolated Fcabs, indicating that this amino acid sequence may also be important for CD137 binding.

As described in the background section above, the initial ligation of the CD137 ligand to its receptor, CD137, initiates a chain of events that leads to trimerization of CD137, followed by receptor clustering, activation of the intracellular NFkB signaling pathway, and subsequent immune cell activation. In order for a therapeutic agent to effectively activate CD137, multiple CD137 monomers must be linked together to mimic a trimeric ligand.

Utomilumab is an IgG2 molecule and its agonist activity is dependent on cross-linking with Fcγ receptors. Urelumab is an IgG4 molecule with constitutive activity and therefore does not require cross-linking with Fcγ receptors for its activity, although its agonist activity is enhanced by cross-linking with some Fcγ receptors. Fcγ receptors are found throughout the human body. Thus, the immune cell activating activity of utomilumab and urelumab is not restricted to specific sites in the body and may therefore be exerted in the liver or elsewhere in the body.

The present inventors have shown that the Fcabs of the present invention require cross-linking to cluster and activate CD137. However, it should be noted that this is not a characteristic feature of the Fcabs that bind CD137. Rather, many of the Fcabs isolated during the screening program bound CD137 but did not require cross-linking to cluster and activate CD137, or induced limited clustering and activation of CD137 in the absence of cross-linking.

As mentioned above, a disadvantage of Fcγ receptor-mediated cross-linking is that Fcγ receptors are found throughout the human body and, accordingly, CD137 activation is not limited to a specific site. Accordingly, the inventors of the present invention introduced mutations into the CH2 domain of Fcab to reduce or eliminate Fcγ receptor binding. Thus, in the absence of cross-linking via an agent other than Fcγ receptors, the Fcabs of the present invention do not exhibit CD137 agonist activity and, accordingly, are not expected to induce liver inflammation.

The inventors have found that the anti-CD137 Fcabs of the invention can be used to generate multispecific, e.g., bispecific, molecules that bind a second antigen in addition to CD137, such as a tumor antigen. Preferably, the multispecific molecule bivalently binds the second antigen, although it is expected that if the second antigen is a cell-associated tumor antigen, monovalent binding of the antigen would be sufficient to cross-link the specific binding member/antibody molecule and induce clustering and activation of CD137. In particular, the inventors have generated antibody molecules comprising the anti-CD137 Fcabs of the invention that bivalently bind a second antigen via their Fab region. The inventors have shown that such bispecific antibody molecules are capable of conditionally activating CD137 in the presence of said second antigen without the need for, e.g., Fc receptor-mediated cross-linking that is required for conventional antibody molecules. It is believed that the binding of antibody molecules to a second antigen causes cross-linking of the antibody molecules at the site of said antigen, which in turn leads to clustering and activation of CD137 on the surface of the T cell. Thus, the agonistic activity of the antibody molecules depends on the presence of both the second antigen and CD137. In other words, the agonistic activity is due to the presence of both antigens. Furthermore, it is believed that cross-linking of antibodies in the presence of a second antigen promotes clustering of CD137 bound via the antigen-binding site of the constant domain of the antibody molecule. If the second antigen is a disease antigen, such as a tumor antigen, it is therefore expected that the antibody molecules will be able to activate immune cells in a disease-dependent manner, such as in the tumor microenvironment. Such targeted activation of immune cells is expected to be beneficial in preventing liver inflammation, as observed with urelumab treatment, as an example.

The present inventors also showed that bispecific antibody molecules comprising the anti-CD137 Fcab of the present invention are capable of suppressing tumor growth in vivo when the second antigen bound by the antibody molecule is an immune cell antigen, a tumor antigen, or an antigen expressed on both tumor cells and immune cells. Furthermore, more effective tumor growth suppression was observed with the bispecific antibody molecules compared to a combination of two monospecific antibody molecules when one of the antibody molecules contained the same constant domain and the other antibody molecule contained the same variable domain binding site as the bispecific molecule, indicating that enhanced clustering and signaling of CD137 and, accordingly, T cell activation and the corresponding antitumor effects are observed when two binding sites are present on the same molecule.

Antibody molecules comprising an anti-CD137 Fcab according to the present invention and a Fab region specific for a second antigen preferably bind bivalently to both CD137 and the second antigen. This is advantageous since bivalent binding of both targets is expected to make the binding between a CD137-expressing T cell and the second antigen more stable, thereby prolonging the time during which the T cell is localized to a particular location, such as a tumor microenvironment, and can act on a disease, such as a tumor. This is in contrast to the vast majority of conventional bispecific antibody formats, which are heterodimeric and bind each target antigen monovalently via one arm of the Fab. Such a monovalent interaction is expected to be not only less stable, but in many cases it will be insufficient to induce clustering of TNF receptors, primarily CD137.

According to an alternative preferred embodiment, the antibody molecule comprises an anti-CD137 Fcab of the present invention capable of bivalently binding CD137 and a monovalent binding site specific for a second antigen, such as a single Fab domain. The monovalent binding site may bind a tumor-associated antigen, as an example. For such molecules, it is expected that monovalent binding of the second antigen will allow for more dense packing of the antibody molecules on the cell surface, resulting in increased clustering of CD137 and, accordingly, T cell activation.

Another feature of the antibody molecules comprising the anti-CD137 Fcab of the present invention is that the two antigen-binding sites for CD137 and the second antigen are both contained within the antibody structure itself. In particular, the antibody molecules do not require fusion of other proteins to the antibody molecule by linkers or other means in order to produce a molecule that bivalently binds to both of its targets. This provides a number of advantages. In particular, the antibody molecules can be produced using methods similar to those used to produce standard antibodies, since they do not contain any additional fusion moieties. The structure is also expected to result in improved antibody stability, since linkers can degrade over time, resulting in a heterogeneous population of antibody molecules. Antibodies in the population that have only one fusion protein may not be able to induce conditional agonism of TNF receptors, such as CD137, as effectively as antibodies that have two fusion proteins. Cleavage/degradation of the linker may occur prior to administration or after administration of the therapeutic agent to the patient (e.g., by enzymatic cleavage or in vivo pH of the patient), resulting in a decrease in its effectiveness during circulation in the patient. Since there are no linkers in antibody molecules, the antibody molecules are expected to retain the same number of binding sites both before and after administration. In addition, the structure of the antibody molecules is also advantageous from the point of view of the immunogenicity of the molecules, since the introduction of fusion proteins or linkers or both can induce immunogenicity upon administration of the molecules to the patient, resulting in a decrease in the effectiveness of the therapeutic agent.

The present inventors have also shown that the rigid positioning and/or close proximity of the antigen-binding sites for CD137, which is due to the rigid structure of the Fcab molecules according to the present invention, is an advantage for inducing clustering of CD137 compared to molecules in which the binding site for CD137 is not an integral part of the antibody structure, but is provided, for example, by linking groups attached, for example, to the antibody molecule, or a part thereof, by flexible linkers.

Thus, according to the present invention, it is proposed:

[1] A specific binding element that binds CD137 and comprises an antigen-binding site for CD137 located within the CH3 domain of said specific binding element, wherein said antigen-binding site for CD137 comprises a first sequence located within the structural loop AB of said CH3 domain, wherein said sequence comprises the sequence PPY (SEQ ID NO: 10).

[2] A specific binding element according to claim [1], wherein said specific binding element comprises an insert in said structural loop AB.

[3] The specific binding element of claim [2], wherein said insert is from 1 to 10 amino acids in length.

[4] The specific binding element of claim [3], wherein said insert is from 4 to 6 amino acids in length.

[5] The specific binding element of claim [4], wherein said insert is 5 amino acids in length.

[6] The specific binding element according to any one of paragraphs [2] to [5], wherein said insert is located between positions 10 and 19 of said CH3 domain of said specific binding element, wherein said amino acid residue numbering corresponds to the ImMunoGeneTics (IMGT) numbering scheme.

[7] The specific binding element according to claim [6], wherein said insert is located between positions 14 and 17 of said CH3 domain of said specific binding element.

[8] The specific binding element according to claim [7], wherein said insert is located between positions 16 and 17 of said CH3 domain of said specific binding element.

[9] The specific binding element according to any one of paragraphs [2] to [8], wherein said insert is located at positions 16.5 to 16.1 of said CH3 domain of said specific binding element, wherein said amino acid residue numbering corresponds to the ImMunoGeneTics (IMGT) numbering scheme.

[10] A specific binding element according to any one of paragraphs [1] to [9], wherein the PPY sequence is located between positions 15 and 17 of said CH3 domain, and wherein said amino acid residue numbering corresponds to the IMGT numbering scheme.

[11] A specific binding element according to any one of claims [1] to [10], wherein the PPY sequence is located between positions 16 and 17 of said CH3 domain, and wherein said amino acid residue numbering corresponds to the IMGT numbering scheme.

[12] The specific binding element according to claim [11], wherein said PPY sequence is located at positions 16.3, 16.2 and 16.1 of said CH3 domain.

[13] The specific binding element according to any one of paragraphs [1] to [12], wherein said first sequence is the first sequence of a specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 138;

(ii) FS22-172-002, represented by SEQ ID NO: 129;

(iii) FS22-172-004, represented by SEQ ID NO: 147;

(iv) FS22-172-001, represented by SEQ ID NO: 120;

(v) FS22-172-005, represented by SEQ ID NO: 156;

(vi) FS22-172-006, as shown in SEQ ID NO: 110; or

(vii) FS22-172, represented by SEQ ID NO: 110.

[14] The specific binding element according to claim [13], wherein said first sequence is the first sequence of the specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 138;

(ii) FS22-172-002, represented by SEQ ID NO: 129;

(iii) FS22-172-004, represented by SEQ ID NO: 147;

(iv) FS22-172-001, represented by SEQ ID NO: 120;

(v) FS22-172-005, as shown in SEQ ID NO: 156; or

(vi) FS22-172-006, represented by SEQ ID NO: 110.

[15] The specific binding element according to claim [14], wherein said first sequence is the first sequence of the specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 138;

(ii) FS22-172-002, as shown in SEQ ID NO: 129; or

(iii) FS22-172-004, represented by SEQ ID NO: 147.

[16] The specific binding element according to claim [15], wherein said first sequence is the first sequence of the specific binding element FS22-172-003, presented in SEQ ID NO: 138.

[17] The specific binding element according to claim [10], wherein said PPY sequence is located at positions 16, 16.5 and 16.4 of said CH3 domain.

[18] The specific binding member according to any one of paragraphs [1] to [10] or [17], wherein said first sequence is the first sequence of the specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably the specific binding member FS22-053-008, presented in SEQ ID NO: 19.

[19] The specific binding element according to any one of paragraphs [13] to [16] or [18], wherein said first sequence is located between positions 14 and 17 of said CH3 domain of said specific binding element, wherein said amino acid residue numbering corresponds to the IMGT numbering scheme.

[20] The specific binding element according to claim [19], wherein said first sequence is located at positions 15, 16, 16.5, 16.4, 16.3, 16.2 and 16.1 of said CH3 domain of said specific binding element.

[21] A specific binding element according to any one of paragraphs [1] to [20], wherein said specific binding element further comprises a second sequence located in the EF structural loop of said CH3 domain.

[22] The specific binding element according to claim [21], wherein said second sequence is the second sequence of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably the specific binding element FS22-172-003, presented in SEQ ID NO: 111.

[23] The specific binding element according to claim [21], wherein said second sequence is a second sequence of the specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 20;

(ii) FS22-053-009, represented by SEQ ID NO: 29;

(iii) FS22-053-011, represented by SEQ ID NO: 47;

(iv) FS22-053-017, represented by SEQ ID NO: 101;

(v) FS22-053-014, represented by SEQ ID NO: 74;

(vi) FS22-053-010, represented by SEQ ID NO: 38;

(vii) FS22-053-012, represented by SEQ ID NO: 56;

(viii) FS22-053-013, represented by SEQ ID NO: 65;

(ix) FS22-053-015, represented by SEQ ID NO: 83;

(x) FS22-053-016, as represented in SEQ ID NO: 92; or

(xi) FS22-053, represented by SEQ ID NO: 174.

[24] The specific binding element according to claim [23], wherein said second sequence is a second sequence of the specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 20;

(ii) FS22-053-009, represented by SEQ ID NO: 29;

(iii) FS22-053-011, represented by SEQ ID NO: 47;

(iv) FS22-053-017, as represented in SEQ ID NO: 101; or

(v) FS22-053-014, represented by SEQ ID NO: 74.

[25] The specific binding element according to claim [24], wherein said second sequence is a second sequence of the specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 20;

(ii) FS22-053-009, represented by SEQ ID NO: 29;

(iii) FS22-053-011, as represented by SEQ ID NO: 47; or

(iv) FS22-053-017, represented by SEQ ID NO: 101.

[26] The specific binding element according to claim [25], wherein said second sequence is the second sequence of the specific binding element FS22-053-008, presented in SEQ ID NO: 20.

[27] The specific binding element according to any one of paragraphs [21] to [26], wherein said second sequence is located at positions 92 to 98 of said CH3 domain of said specific binding element, wherein said amino acid residue numbering corresponds to the IMGT numbering scheme.

[28] A specific binding element according to any one of paragraphs [1] to [27], wherein said specific binding element further comprises a third sequence located in the structural loop CD of said CH3 domain.

[29] The specific binding element according to claim [28], wherein said third sequence is located at positions 43 to 78 of said specific binding element, wherein said numbering of amino acid residues corresponds to the IMGT numbering scheme.

[30] The specific binding element according to any one of paragraphs [28]-[29], wherein said third sequence has the sequence presented in SEQ ID NO: 2.

[31] The specific binding element according to any one of claims [1] to [30], wherein said CH3 domain is a CH3 domain of human IgG1.

[32] The specific binding element according to any one of paragraphs [1]-[16], [19]-[22] and [27]-[31], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 139;

(ii) FS22-172-002, represented by SEQ ID NO: 130;

(iii) FS22-172-004, represented by SEQ ID NO: 148;

(iv) FS22-172-001, represented by SEQ ID NO: 121;

(v) FS22-172-005, represented by SEQ ID NO: 157;

(vi) FS22-172-006, as represented by SEQ ID NO: 165; or

(vii) FS22-172, represented by SEQ ID NO: 112.

[33] The specific binding element according to claim [32], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 139;

(ii) FS22-172-002, represented by SEQ ID NO: 130;

(iii) FS22-172-004, represented by SEQ ID NO: 148;

(iv) FS22-172-001, represented by SEQ ID NO: 121;

(v) FS22-172-005, as shown in SEQ ID NO: 157; or

(vi) FS22-172-006, represented by SEQ ID NO: 165.

[34] The specific binding element according to claim [33], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-172-003, represented by SEQ ID NO: 139;

(ii) FS22-172-002, as shown in SEQ ID NO: 130; or

(iii) FS22-172-004, represented by SEQ ID NO: 148.

[35] The specific binding element according to claim [34], wherein said specific binding element comprises the CH3 domain sequence of the specific binding element FS22-172-003, presented in SEQ ID NO: 139.

[36] The specific binding element according to any one of paragraphs [1]-[10], [17]-[18], [19]-[21] and [23]-[31], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 21;

(ii) FS22-053-009, represented by SEQ ID NO: 30;

(iii) FS22-053-011, represented by SEQ ID NO: 48;

(iv) FS22-053-017, represented by SEQ ID NO: 102;

(v) FS22-053-014, represented by SEQ ID NO: 75;

(vi) FS22-053-010, represented by SEQ ID NO: 39;

(vii) FS22-053-012, represented by SEQ ID NO: 57;

(viii) FS22-053-013, represented by SEQ ID NO: 66;

(ix) FS22-053-015, represented by SEQ ID NO: 84;

(x) FS22-053-016, as represented in SEQ ID NO: 93; or

(xi) FS22-053, represented by SEQ ID NO: 175.

[37] The specific binding element according to claim [36], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 21;

(ii) FS22-053-009, represented by SEQ ID NO: 30;

(iii) FS22-053-011, represented by SEQ ID NO: 48;

(iv) FS22-053-017, as represented in SEQ ID NO: 102; or

(v) FS22-053-014, represented by SEQ ID NO: 75.

[38] The specific binding element according to claim [37], wherein said specific binding element comprises a CH3 domain sequence of a specific binding element:

(i) FS22-053-008, represented by SEQ ID NO: 21;

(ii) FS22-053-009, represented by SEQ ID NO: 30;

(iii) FS22-053-011, as shown in SEQ ID NO: 48; or

(iv) FS22-053-017, represented by SEQ ID NO: 102.

[39] The specific binding element according to claim [38], wherein said specific binding element comprises the CH3 domain sequence of the specific binding element FS22-053-008, as presented in SEQ ID NO: 21.

[40] The specific binding element according to claim [37], wherein said specific binding element comprises the CH3 domain sequence of the specific binding element FS22-053-017, as presented in SEQ ID NO: 102.

[41] The specific binding element according to claim [37], wherein said specific binding element comprises the CH3 domain sequence of the specific binding element FS22-053-014, presented in SEQ ID NO: 75.

[42] A specific binding member according to any one of paragraphs [1] to [41], wherein said specific binding member further comprises a CH2 domain, preferably a CH2 domain of human IgG1.

[43] The specific binding element according to any one of paragraphs [1] to [42], wherein said specific binding element is a dimer of two identical polypeptide chains, each of which comprises a CH2 and CH3 domain.

[44] The specific binding element according to [42] or [43], wherein said CH2 domain has the sequence presented in SEQ ID NO: 6 or 5.

[45] The specific binding element according to any one of claims [42] to [44], further comprising an immunoglobulin hinge region, or a portion thereof, at the N-terminus of said CH2 domain, preferably a human IgG1 hinge region, or a portion thereof.

[46] The specific binding element according to claim [45], wherein said hinge region has the sequence presented in SEQ ID NO: 179, or a fragment thereof.

[47] The specific binding element according to claim [46], wherein said hinge region has the sequence presented in SEQ ID NO: 7.

[48] The specific binding element according to any one of paragraphs [1]-[16], [19]-[22], [27]-[35] and [42]-[47], wherein said specific binding element comprises the sequence of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172 presented in SEQ ID NO: 141, 132, 150, 123, 159, 167 and 114, respectively.

[49] The specific binding element according to claim [48], wherein said specific binding element comprises the sequence of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, presented in SEQ ID NO: 141, 132, 150, 123, 159 and 167, respectively.

[50] The specific binding element according to claim [49], wherein said specific binding element comprises the sequence of the specific binding element FS22-172-003, FS22-172-002 or FS22-172-004, presented in SEQ ID NO: 141, 132 and 150, respectively.

[51] The specific binding element according to claim [50], wherein said specific binding element comprises the sequence of the specific binding element FS22-172-003 presented in SEQ ID NO: 141.

[52] A specific binding element according to any one of claims [1]-[10], [17]-[18], [19]-[21], [23]-[31] and [36]-[47], wherein said specific binding element comprises the sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053 as set forth in SEQ ID NO: 23, 32, 50, 104, 77, 41, 59, 68, 86, 95 and 175, respectively.

[53] The specific binding element according to claim [52], wherein said specific binding element comprises the sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, as presented in SEQ ID NO: 23, 32, 50, 104 and 77, respectively.

[54] The specific binding element according to claim [53], wherein said specific binding element comprises the sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, as presented in SEQ ID NO: 23, 32, 50 and 103, respectively.

[55] The specific binding element according to claim [54], wherein said specific binding element comprises the sequence of the specific binding element FS22-053-008 presented in SEQ ID NO: 23.

[56] A specific binding member according to any one of paragraphs [1] to [55], wherein said specific binding member binds human CD137.

[57] The specific binding element of claim [56], wherein said human CD137 has, comprises or consists of the sequence set forth in SEQ ID NO: 181.

[58] The specific binding member of any one of paragraphs [1] to [55], wherein said specific binding member binds to dimeric CD137 with higher affinity than to monomeric CD137.

[59] A specific binding member according to any one of paragraphs [1] to [58], wherein said specific binding member binds cynomolgus monkey CD137.

[60] A specific binding element or antibody molecule according to claim [59], which is characterized in that said cynomolgus monkey CD137 has, comprises or consists of the sequence presented in SEQ ID NO: 183.

[61] The specific binding element according to any one of paragraphs [1] to [60], wherein said specific binding element further comprises a second antigen-binding site.

[62] The specific binding element according to claim [61], wherein the specific binding element is a multispecific molecule.

[63] The specific binding member of claim [62], wherein the specific binding member is a bispecific, trispecific or tetraspecific molecule.

[64] The specific binding element according to claim [63], wherein the specific binding element is a bispecific molecule.

[65] The specific binding element according to any one of paragraphs [61] to [64], wherein said second antigen-binding site is a CDR-based antigen-binding site.

[66] The specific binding element according to claim [65], wherein said second antigen-binding site comprises CDR1, CDR2 and CDR3 of the variable domain of the heavy chain and CDR1, CDR2 and CDR3 of the variable domain of the light chain.

[67] The specific binding element according to any one of paragraphs [61]-[66], wherein said second antigen-binding site comprises a heavy chain variable domain and a light chain variable domain.

[68] The specific binding member according to any one of paragraphs [61] to [67], wherein said specific binding member is an antibody molecule.

[69] The antibody molecule of claim [68], wherein said antibody molecule is a human IgG1 molecule.

[70] An antibody molecule according to any one of paragraphs [65]-[69], wherein said CDR-based antigen-binding site of said antibody molecule binds a second antigen selected from the group consisting of: an immune cell antigen and a disease antigen.

[71] An antibody molecule according to claim [70], wherein said disease antigen is a tumor antigen or a pathogen antigen.

[72] The antibody molecule of claim [70], wherein said immune cell antigen is an immune regulatory molecule such as PD-L1.

[73] An antibody molecule according to claim [70], wherein said immune cell antigen is a member of the tumor necrosis factor receptor superfamily (TNFRSF).

[74] An antibody molecule according to claim [71], wherein said tumor antigen is a tumor-associated antigen (TAA).

[75] An antibody molecule according to [71] or [74], wherein said tumor antigen is a cell surface antigen on a cancer cell.

[76] The antibody molecule of claim [71], wherein said tumor antigen is a soluble multimer.

[77] The antibody molecule of claim [76], wherein the soluble multimer is at least a dimer.

[78] The antibody molecule of claim [77], wherein the soluble multimer is at least a trimer.

[79] The antibody molecule of claim [71], wherein said pathogen antigen is a bacterial or viral antigen.

[80] An antibody molecule according to any one of paragraphs [70]-[79], wherein said antibody molecule is capable of activating CD137 present on an immune cell in the presence of said second antigen.

[81] An antibody molecule according to any one of paragraphs [70]-[80], wherein binding of said antibody molecule to CD137 and said second antigen causes clustering of CD137 on an immune cell.

[82] An antibody molecule according to [80] or [81], wherein said immune cell is a T cell.

[83] The specific binding member or antibody molecule of any one of paragraphs [1] to [82], wherein said specific binding member or antibody molecule is modified to reduce or eliminate binding of said CH2 domain of said specific binding member or antibody molecule to one or more Fcγ receptors.

[84] The specific binding member or antibody molecule of any one of paragraphs [1] to [83], wherein said specific binding member or antibody molecule does not activate Fcγ receptors.

[85] The specific binding element or antibody molecule of claim [83] or [84], wherein said Fcγ receptor is selected from the group consisting of: FcγRI, FcγRIIa, FcγRIIb and FcγRIII.

[86] A specific binding element or antibody molecule according to any one of paragraphs [1] to [85], wherein said specific binding element or antibody molecule is conjugated to a bioactive molecule.

[87] The specific binding member or antibody molecule of any one of paragraphs [1] to [85], wherein said specific binding member or antibody molecule is conjugated to a detectable label.

[88] A nucleic acid molecule encoding a specific binding element or an antibody molecule according to any one of paragraphs [1] to [85].

[89] A nucleic acid molecule according to claim [88], wherein said nucleic acid molecule(s) comprises(contain):

(i) the nucleic acid sequence of the CH3 domain of the specific binding member of FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053 as set forth in SEQ ID NOs: 22, 31, 49, 103, 76, 40, 58, 67, 85, 94 and 176, respectively; or

(ii) the nucleic acid sequence of the CH3 domain of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172 as set forth in SEQ ID NOs: 140, 131, 149, 122, 158, 166 and 113, respectively.

[90] A nucleic acid molecule according to [88] or [89], wherein said nucleic acid molecule comprises a nucleic acid sequence: of a specific binding element:

(i) FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053 as depicted in SEQ ID NOs: 24, 33, 51, 105, 78, 42, 60, 69, 87, 96 and 177, respectively; or

(ii) FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172 as shown in SEQ ID NO: 142, 133, 151, 124, 160, 168 and 115, respectively.

[91] A nucleic acid molecule according to [89] or [90], wherein said nucleic acid molecule comprises a CH3 domain nucleic acid sequence or a specific binding element nucleic acid sequence:

(i) FS22-172-003; or

(ii) FS22-053-008.

[92] A vector comprising a nucleic acid according to any one of paragraphs [88]-[91].

[93] A recombinant host cell comprising a nucleic acid according to any one of claims [88] to [91], or a vector according to claim [92].

[94] A method for producing a specific binding element or antibody molecule according to any one of paragraphs [1] to [85], comprising culturing a recombinant host cell according to paragraph [93] under conditions for producing said specific binding element or antibody molecule.

[95] The method according to item [94], further comprising isolating and/or purifying said specific binding element or antibody molecule.

[96] A pharmaceutical composition comprising a specific binding element or an antibody molecule according to any one of paragraphs [1]-[87] and a pharmaceutically acceptable excipient.

[97] A specific binding element or antibody molecule according to any one of paragraphs [1] to [87] for use in a method of treating a human or animal organism by therapy.

[98] A method of treating a disease or disorder in an individual comprising administering to said individual a therapeutically effective amount of a specific binding member or antibody molecule according to any one of paragraphs [1] to [87].

[99] A specific binding member or antibody molecule for use according to claim [97], or a method according to claim [98], wherein said treatment is the treatment of cancer or an infectious disease in an individual.

[10] A specific binding member or antibody molecule for use according to [97] or [99], or a method according to [98] or [99], wherein said method of treatment comprises administering said specific binding member or antibody molecule to an individual in combination with a second therapeutic agent.

Brief description of the drawings

Figure 1: Figures 1A, B and C show the sequence alignment of the CH3 domains of Fcab FS22-053, FS22-053-008, FS22-053-009, FS22-053-010, FS22-053-011, FS22-053-012, FS22-053-013, FS22-053-014, FS22-053-015, FS22-053-016, FS22-053-017, FS22-172, FS22-172-001, FS22-172-002, FS22-172-003, FS22-172-004, FS22-172-005 and FS22-172-006, as well as wild-type (WT) Fcab. Residue numbers are given according to the IMGT, IMGT exon (sequential numbering), EC, and Kabat numbering systems. Figure 1D shows the percent sequence identity of the CH3 domain of clones FS22-053-008 through FS22-053-016 and FS22-053-017 (see Example 10.1) compared to the CH3 domain of parental FS22-053. Figure 1E shows the percent sequence identity of the CH3 domain of clones FS22-172-001 through FS22-172-006 compared to the CH3 domain of parental FS22-053.

Figure 2 shows NF-kB signaling determined from luminescence values as a measure of luciferase production induced by CD137 clustering and activation. Parental human CD137 Fcabs FS22-053 and FS22-172 in the HelD1.3 mock mAb ² format trigger limited CD137 clustering and NF-kB signaling when cross-linked to the L protein in the HEK hCD137 NF-kB reporter assay (open and filled circles). Affinity-matured human anti-CD137 Fcabs FS22-053-008, -009, -011 (open squares, triangles and diamonds) and FS22-172-002, -003, -004 (filled squares, triangles and diamonds) in the HelD1.3 mock mAb ² format trigger stronger CD137 clustering and NF-kB signaling upon cross-linking to protein L in the HEK hCD137 NF-kB reporter assay compared to the parental clones. The positive control anti-human CD137 mAb, 20H4.9, showed increased luminescence upon cross-linking to protein L (dashed line) with a lower _EC50 value compared to affinity-matured human anti-CD137 Fcabs.

Figure 3 shows the release of IL-2 in the T cell activation assay in the presence of Fcab to human CD137. FS22-053-008, FS22-053-009, FS22-053-011 and FS22-172-002, FS22-172-003, FS22-172-004 in the format of mAb ² to PD-L1 (S70 LALA) trigger clustering and activation of CD137 only when cross-linked to HEK cells overexpressing PD-L1 (empty and filled black symbols), resulting in the release of mouse interleukin-2 (mIL-2) in the DO11.10 T cell activation assay. At increasing concentrations, the positive control anti-human CD137 mAb, 20H4.9, showed an increase in mIL-2 release (dashed line), however, the maximal release was significantly lower than that of anti-human CD137 Fcab. All anti-human CD137 Fcab in the anti-PD-L1 mAb ² model format without HEK cells overexpressing PD-L1 for cross-linking (filled gray symbols) showed significantly lower mIL-2 release compared to cross-linking (empty and filled black symbols).

Figure 4 shows the release of human IL-2 (hIL-2) in a primary human CD8 ⁺ T cell activation assay. Affinity-matured human anti-CD137 Fcabs FS22-053-008, FS22-053-009, FS22-053-011, FS22-172-002, FS22-172-003, and FS22-172-004 in the format of PD-L1 mAb ² triggered the activation of CD137 CD8 ⁺ T cells when mAb ² was cross-linked with PD-L1-overexpressing HEK cells, resulting in the release of human IL-2. The activity of the affinity matured Fcabs was greater than that of the parental Fcabs, FS22-053 (open circles) and FS22-172 (filled circles). The positive control anti-human CD137 mAb, 20H4.9 (dashed line), showed an increase in hIL-2 release that was greater than that of the Fcabs, but the _EC50 value was greater than that of some anti-human CD137 Fcabs, indicating that some anti-human CD137 Fcabs achieved better activity.

Figure 5 shows NF-kB signaling determined from luminescence values as a measure of luciferase production induced by CD137 clustering and activation. The mouse anti-CD137 Fcab FS22m-063 in the HelD1.3 mock mAb ² format triggered CD137 clustering and NF-kB signaling when cross-linked to protein L in the HEK mCD137 NF-kB reporter assay (filled circles). At concentrations above 100 nM, and in the absence of cross-linking to protein L, FS22m-063 triggered some CD137 clustering and NF-kB signaling, but at levels much lower than when cross-linked (open circles). The positive control anti-mouse CD137 mAb, Lob12.3, showed increased luminescence upon cross-linking to protein L (filled squares) compared to no cross-linking (open squares), but the maximal response achieved at the highest antibody concentration was significantly less than that of anti-mouse CD137 Fcab. As expected, the isotype controls showed no activity.

Figure 6 shows tumor volume measurements of the syngeneic MC38 tumor model grown subcutaneously in C57BL6 mice treated with G1-AA/4420 (IgG control), G1-AA/S70 (PD-L1 positive control), G1-AA/Lob12.3 (CD137 positive control), a combination of G1-AA/S70 plus G1-AA/Lob12.3, and FS22m-063-AA/S70 (FS22m-063 mouse anti-CD137 Fcab in anti-PD-L1 mAb ² format). Mean tumor volume [in mm ³ ] plus or minus 95% confidence interval is shown. FS22m-063-AA/S70 was able to significantly reduce tumor growth in the syngeneic MC38 tumor model compared to control IgG-treated mice, positive control anti-PD-L1 mAb-treated mice, and positive control anti-CD137-treated mice. Statistical significance is shown pairwise for growth rates over the entire study time using mixed model analysis. ****≤0.0001 p-value.

Figure 7: A shows tumor volume measurements of a syngeneic MC38 tumor model grown subcutaneously in C57BL/6 mice treated with G1-AA/4420 (IgG control), G1-AA/F2 (PD-1 positive control), G1/Lob12.3 (CD137 positive control), a combination of G1-AA/F2 plus G1-AA/Lob12.3, and FS22m-063-AA/F2 (FS22m-063 mouse anti-CD137 Fcab in anti-PD-1 mAb ² format). The mean tumor volume [in mm ³ ] plus or minus 95% confidence interval is shown. FS22m-063-AA/F2 was able to significantly reduce tumor growth in the syngeneic MC38 tumor model compared to mice treated with control IgG. Statistical significance is shown pairwise for growth rates over the entire study duration using mixed model analysis. ****≤0.0001 p-value. B shows the same data as Figure 7A except that tumor volume in each treated mouse is shown separately. This indicates that, with the exception of one mouse, none of the FS22m-063-AA/F2-treated mice showed any tumor regrowth over the course of the study. In contrast, tumor regrowth was observed in G1-AA/Lob12.3- and G1-AA/F2-treated mice by the end of the study.

Figure 8 shows the mean tumor volume measurements in a mesothelin-positive subcutaneous syngeneic CT26 tumor model (Balb/c). Mice received either G1-AA/4420 (IgG control) or the FS22m-063-AA/FS28m-228 mAb². Mice receiving FS22m-063-AA/FS28m-228 showed significant inhibition of tumor growth compared to mice receiving control IgG. The mean tumor volume [in ^mm3 ] plus or minus 95% confidence interval is plotted. Statistical significance is shown by pairwise comparison of growth rates over the entire study time using mixed model analysis. ****≤0.0001 p-value.

Figure 9 shows individual tumor volume measurements in the syngeneic CT26.G10 tumor model treated with G1-AA/HelD1.3 (human IgG1 control), FS22m-063-AA/HelD1.3 (anti-mouse CD137 Fcab in mock mAb ² format), FS22m-063-AA/4420 (anti-mouse CD137 Fcab in mock mAb ² format), G1-AA/FS28m-228-010 (anti-mouse MSLN mAb), a combination of FS22m-063-AA/HelD1.3 and G1-AA/FS28m-228-010 (anti-mouse CD137 Fcab plus anti-mouse MSLN mAb), and FS22m-063-AA/FS28m-228-010 (anti-mouse CD137 Fcab plus anti ^- mouse MSLN mAb) CD137/MSLN mice). FS22m-063-AA/FS28m-228-010 showed reduced tumor growth compared to isotype control as well as other treatment groups.

Figure 10 shows the Kaplan-Meier survival curve for the syngeneic CT26.G10 tumor model treated with G1-AA/HelD1.3 (IgG control), FS22m-063-AA/HelD1.3 (mouse anti-CD137 Fcab in MAb ² format), FS22m-063-AA/4420 (mouse anti-CD137 Fcab in MAb ² format), G1-AA/FS28m-228-010 (mouse anti-MSLN Fab), a combination of FS22m-063-AA/HelD1.3 and G1-AA/FS28m-228-010 (mouse anti-CD137 Fcab plus mouse anti-MSLN Fab), and FS22m-063-AA/FS28m-228-010 (MAb ² to CD137/MSLN mice). The results show that treatment with FS22m-063-AA/FS28m-228-010 resulted in significantly improved survival compared to mice receiving isotype control or other treatment groups.

Figure 11 shows the release of mouse IL-2 in the DO11.10 human CD137-expressing T cell activation assay testing Fcabs FS22-053-014 and FS22-053-017, cross-reacting with mouse and human CD137, in a mock mAb ² format when cross-linked with protein L. Both Fcabs were able to activate CD137 when cross-linked with protein L. The positive control anti-human CD137 mAb, 20H4.9, showed an increase in mIL-2 release, consistent with mock mAb ² FS22-053-017. Both anti-human CD137 Fcabs in a mock mAb ² format without protein L for cross-linking (empty symbols) showed little mIL-2 release compared to cross-linking (filled symbols).

Figure 12 shows the release of mouse IL-2 in the DO11.10 mouse CD137-expressing T cell activation assay testing Fcabs FS22-053-014 and FS22-053-017, cross-reacting with mouse and human CD137, in a mock mAb ² format when cross-linked to protein L. FS22-053-014 and FS22-053-017, as well as the anti-mouse CD137 Fcab FS22m-063, all in the HelD1.3 mock mAb ² format, activated CD137 when cross-linked to protein L, resulting in the release of mIL-2. The positive control anti-mouse CD137 mAb, Lob12.3, showed increased mIL-2 release as expected. All anti-CD137 Fcabs in the mock mAb ² format without protein L for cross-linking (dashed lines) showed significantly reduced mIL-2 release compared to cross-linking. In this assay, FS22-053-017 had lower activity ( _EC50 8-fold worse compared to FS22-053-014) but still showed activity in the assay.

Figure 13 shows the homology models of Fcab FS22 compared to a structural matrix (PDB ID 5JII) containing the wild-type CH3 domain of human IgG1 Fc. The images show Fc 5JII (A) and the representative structures of Fcab FS22-053-008 (B) and FS-172-003 (C). One CH3 domain of each structure was visualized using a gray ribbon diagram image. The AB, CD, and EF loops are indicated by black shading of the ribbons and labeled accordingly. The predicted structure of the AB loop is highlighted with dotted lines for comparison purposes and shows a protrusion caused by the PPY motif, which likely plays a key role in binding.

Figure 14 shows a sensorgram of the binding kinetics of homodimeric Fcab (FS22-172-003-AA) in MAT² format (i.e., containing 2 CD137-binding CH3 domains) compared to heterodimeric Fcab in MAT² format (containing 1 CD137-binding CH3 domain and one wild-type CH3 domain). This sensorgram shows that both molecules bound to CD137 even through only one binding strand. The dissociation rates were different, with homodimeric Fcab showing a much slower dissociation profile compared to heterodimeric Fcab. These sensorgrams demonstrate that Fcab to CD137 can bivalently bind CD137.

Figure 15 shows binding of Fcab to human CD137 in mock MAb² format (FS22-172-003-AA/HelD1.3), positive control antibody (G1-AA/20H4.9), and isotype control antibody (G1-AA/HelD1.3) to cells expressing a range of levels of CD137, including a negative control line (hCD137 ^neg DO11.10) (A). Binding was determined by flow cytometry. The results showed that while G1-AA/20H4.9 was able to bind all cell lines expressing CD137 (BE), FS22-172-003-AA/HelD1.3 did not bind the DO11.10 cell line expressing low levels of CD137 (B), and that the fold difference in binding compared to the positive control was inversely proportional to the amount of CD137 expressed by the cell line.

Detailed description

The present invention relates to specific binding members that bind CD137. CD137 is also known as a member of the tumor necrosis factor receptor superfamily 9 (TNFRSF9) or 4-1BB. The specific binding member preferably binds human CD137, more preferably human and cynomolgus monkey CD137, even more preferably dimeric human and cynomolgus monkey CD137. The portion of CD137 bound by the specific binding member is preferably an extracellular domain of CD137. The extracellular domain of human and cynomolgus monkey CD137 may comprise or consist of the sequence presented in SEQ ID NOs 181 and 183, respectively. The specific binding member is preferably capable of binding to CD137 expressed on the surface of a cell. The cell is preferably an immune cell, such as a CD8 ⁺ or CD4 ⁺ T cell or a regulatory T cell (Treg), preferably a CD8 ⁺ T cell, or a B cell, a natural killer cell (NK cell), a natural killer T cell (NKT cell), a dendritic cell (DC), or a tumor infiltrating lymphocyte (TIL).

The specific binding member preferably specifically binds CD137. The term "specific" may refer to a situation in which the specific binding member will not show any significant binding to molecules other than its specific binding partner(s), in this case CD137. The term "specific" is also applicable when the specific binding member is specific for particular epitopes, such as epitopes on CD137 carried by a number of antigens, in which case the specific binding member will be able to bind to a variety of antigens bearing the epitope. The specific binding member preferably does not bind or does not show any significant binding to CD40, OX40 and/or GITR.

As explained in the background section above, treatment of patients with the CD137 antibody urelumab was associated with dose-limiting, severe liver inflammation. Without being bound by theory, it is believed that the liver inflammation observed with urelumab treatment could be caused by activation of T cells present in the liver or infiltration and accumulation of activated T cells in the liver of patients. In order to select molecules that cause reduced or no liver inflammation, the inventors selected Fcabs with high avidity for CD137. In particular, the inventors selected Fcabs that bound dimeric CD137 with higher affinity than monomeric CD137. CD137 expression by T cells is upregulated upon priming and activation. It is believed that due to the higher expression of CD137 on activated T cells, CD137 will be in the form of dimers, trimers, and higher order multimers on the surface of such cells. In contrast, CD137 expression on inactive T cells is low or even undetectable. Thus, it is believed that CD137, to the extent that it is expressed at all on the surface of such T cells, is likely to be in the monomeric form. Thus, Fcabs that bind to CD137 with high avidity are believed to preferentially bind to activated T cells as opposed to inactive T cells, such as inactive T cells present in the liver, and therefore exhibit reduced or absent liver inflammation.

The specific binding member preferentially binds to dimeric human CD137 with an affinity (K _D ) of 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 5 nM, 4 nM, 3 nM, or 2 nM, or with a higher affinity.

According to a preferred embodiment, the specific binding member binds to dimeric CD137 with a higher affinity than to monomeric CD137. According to a preferred embodiment, the specific binding member binds to dimeric CD137 with an affinity that is at least 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 110-fold, 120-fold, 130-fold, 140-fold, 150-fold, 160-fold, 170-fold, or 200-fold higher than the affinity of the specific binding member for monomeric CD137.

Human CD137, for example, may have the sequence shown in SEQ ID NO: 183. The sequence is identical regardless of whether the antigen is in monomeric or dimeric form.

Specific binding elements from the FS22-53 and FS22-172 lines were also shown to bind dimeric cynomolgus monkey CD137. Binding to cynomolgus monkey CD137 as well as human CD137 is advantageous because it allows testing of the efficacy and toxicity of the specific binding element in cynomolgus monkeys prior to administration to humans.

According to a preferred embodiment, the specific binding member may bind to dimeric cynomolgus monkey CD137 with an affinity ( _KD ) of 250 nM, 200 nM, 150 nM, 140 nM, 120 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 20 nM, 10 nM, 5 nM, 4 nM, 3 nM or 2 nM or with higher affinity. Preferably, the specific binding member binds to cynomolgus monkey CD137 with an affinity ( _KD ) of 2 nM or with higher affinity.

The specific binding member can bind to dimeric human CD137 and dimeric cynomolgus monkey CD137 with similar affinity. This is believed to be favorable for conducting efficacy and toxicity studies using the specific binding member in cynomolgus monkeys, which may have predictive value for the efficacy and toxicity of the specific binding member in humans.

Thus, according to a preferred embodiment, the specific binding member binds to dimeric cynomolgus monkey CD137 with an affinity that is no more than 10-fold, preferably no more than 5-fold, lower or higher than the affinity with which the specific binding member binds dimeric human CD137.

The specific binding member can bind to dimeric mouse CD137, dimeric cynomolgus monkey CD137, and dimeric human CD137. This is believed to be advantageous because the same specific binding member can be used to perform initial efficacy studies in mice, toxicity studies in cynomolgus monkeys, and clinical studies in humans, potentially simplifying the path to the clinic. In particular, the specific binding members FS22-053-014 and FS22-053-017 were unexpectedly found to bind to dimeric mouse, human, and cynomolgus monkey CD137.

The binding affinity of a specific binding element for a cognate antigen, such as human or cynomolgus monkey CD137, can be determined using surface plasmon resonance (SPR), such as Biacore.

The term "specific binding member" describes an immunoglobulin or fragment thereof comprising a constant domain comprising an antigen-binding site for CD137. In the present application, the term "specific binding member" thus includes antigen-binding fragments, provided that said antigen-binding fragments comprise an antigen-binding site for CD137 located in the constant domain of the specific binding member. The constant domain may be a CL, CH1, CH2, CH3 or CH4 domain, preferably the constant domain is a CH1, CH2 or CH3 domain, more preferably a CH2 or CH3 domain, most preferably a CH3 domain. The specific binding member may be partially or completely synthetically produced.

Preferably, the specific binding member comprises a CH2 and a CH3 domain, wherein said CH2 or CH3 domain, preferably the CH3 domain, comprises an antigen-binding site for CD137. The specific binding member is preferably a dimer of two (identical) polypeptide chains, each comprising a CH2 and a CH3 domain. According to a preferred embodiment, the specific binding member further comprises an immunoglobulin hinge region, or a portion thereof, at the N-terminus of the CH2 domain. Such a molecule is also referred to herein as an antigen-binding Fc fragment or Fcab™. The hinge region may consist of or comprise the sequence of SEQ ID NO: 179 or a fragment thereof. Preferably, the fragment is a C-terminal fragment of the sequence presented in SEQ ID NO: 179. The fragment may be up to 20, up to 10, up to 8 or up to 6 amino acids in length. The fragment may be at least 3, at least 4, at least 5 or at least 6 amino acids in length. According to a preferred embodiment, the hinge region has the sequence presented in SEQ ID NO: 7.

According to a preferred embodiment, the specific binding element is an antibody molecule, preferably a monoclonal antibody, or a fragment thereof. The antibody molecule is preferably human or humanized. The antibody molecule may be an immunoglobulin G molecule, such as an IgG1, IgG2, IgG3 or IgG4 molecule, preferably an IgG1, IgG2 or IgG4 molecule, more preferably an IgG1 molecule, or a fragment thereof.

Since antibodies can be modified in a variety of ways, the term "antibody molecule" should be understood to encompass antibody fragments, derivatives, functional equivalents, and homologues of antibodies, whether natural or wholly or partly synthetic. An example of an antibody fragment containing a CH3 domain is the Fc domain of an antibody. An example of an antibody fragment containing both CDR sequences and a CH3 domain is a minibody that contains an scFv fused to a CH3 domain (Hu et al. (1996) Cancer Res., 56(13):3055-61).

The specific binding member comprises an antigen-binding site for CD137. The antigen-binding site for CD137 is located in the constant domain of the specific binding member, preferably in the CH3 domain. The antigen-binding site for CD137 comprises one or more modified structural loops in the constant domain of the specific binding member. Engineering structural loops of the constant domain of an antibody to create antigen-binding sites for target antigens is known in the art and is described, for example, by Wozniak-Knopp G et al. (2010); WO2006/072620 and WO2009/132876.

Following an extensive selection and affinity maturation program, we isolated two panels of Fcabs that preferentially bound dimeric rather than monomeric human CD137. Surprisingly, all of the isolated Fcabs contained the PPY sequence as well as a five-amino acid insertion in their AB structural loop. The two panels of Fcabs were selected independently from different selection campaigns, each using a library containing the five-amino acid insertion in the AB structural loop. Thus, the PPY sequence was independently selected twice, indicating the importance of this sequence for CD137 binding. Anti-CD137 Fcabs isolated from Fcab libraries that did not contain the five-amino acid insertion in the AB structural loop were not used further, for example because the selected Fcabs could not be affinity maturated. This indicates that the amino acid insert present in the AB structural loop may also be important for CD137 binding. Thus, the presence of a PPY sequence and a 5-amino acid insert in the AB structural loop, wherein said PPY sequence may not necessarily be present entirely or partially within the 5-amino acid insert, may be important for CD137 binding.

Thus, the antigen-binding site for the CD137 specific binding element may comprise a first and/or second sequence, preferably a first and second sequence, wherein said first and second sequences are located in the structural loops AB and EF of the constant domain, preferably the CH3 domain, of the specific binding element, respectively.

According to a preferred embodiment, residues at positions 95 and 96 of the CH3 domain of the specific binding element are wild type, i.e., preferably arginine (R) and tryptophan (W), respectively. Both of these residues are located in the structural loop EF. In the present application, amino acid residue positions are numbered according to the ImMunoGeneTics (IMGT) numbering scheme, unless otherwise indicated. The IMGT numbering scheme is described in Lefranc et al., 2005.

The first sequence preferably comprises the sequence PPY (SEQ ID NO: 10).

The PPY sequence may be located between positions 10 and 19, preferably positions 15 and 17, of the CH3 domain of the specific binding element. According to a preferred embodiment, the PPY sequence is located at positions 16, 16.5 and 16.4 of the CH3 domain. Alternatively, the PPY sequence may be located between positions 16 and 17 of the CH3 domain. According to an alternative preferred embodiment, the PPY sequence is located at positions 16.3, 16.2 and 16.1 of the CH3 domain. In the IMGT numbering scheme, inserted residues are numbered according to the direction of the loop in which they are located. If the loop is "up", the inserted residues take the number of the residue immediately preceding the insertion, with the number of the inserted residue in the sequence being indicated by an increasing decimal number, e.g., 16, 16.1, 16.2, 16.3, when three mutations are present after residue 16. If the loop is "down", the inserted residues take the number of the residue immediately preceding the insertion, with the number of the inserted residue in the sequence being indicated by a decreasing decimal number, e.g., 16, 16.3, 16.2, 16.1, when, as mentioned, three mutations are present after residue 16 (LeFranc et al., 2005, and LeFranc et al. 2015).

According to a preferred embodiment, the structural loop AB comprises an amino acid insert. The insert may be 1 to 10, 2 to 9, 3 to 7, 4 to 6 or 5 amino acids in length. Preferably, the insert is 5 amino acids in length.

The insert may be located between positions 10 and 19, preferably between positions 14 and 17, more preferably between positions 16 and 17 of the CH3 domain of the specific binding element. According to a preferred embodiment, the insert is located at positions 16.5 to 16.1 of the CH3 domain of the specific binding element, as shown in Figure 1.

Most of the specific binding elements identified after affinity maturation contained a leucine (L) residue at position 97 of the CH3 domain. Many of the specific binding elements also contained an aspartic acid (D) residue or a glutamic acid (E) residue at positions 98 of the CH3 domain of the specific binding element. Both of these amino acid substitutions are located within the EF structural loop. These results suggest that one or both of these residues may be important for CD137 binding. Thus, the second sequence preferably comprises an LD or LE sequence, wherein said LD or LE sequence is preferably located at positions 97 and 98 of the CH3 domain of the specific binding element.

The first sequence and the second sequence may be the first and second sequence of the CH3 domain: of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, more preferably of the specific binding element FS22-172-003, FS22-172-002 or FS22-172-004, yet more preferably the specific binding element FS22-172-003.

Alternatively, the first sequence and the second sequence may be the first and second sequence of the CH3 domain: specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015 or FS22-053-016, or FS22-053, preferably specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably specific binding element FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably specific binding element FS22-053-008. According to an alternative preferred embodiment, the first sequence and the second sequence are the first and second sequence of the CH3 domain of specific binding element FS22-053-017 or FS22-053-014, more preferably specific binding element FS22-053-017.

The sequence of the CH3 domain of the specific binding element of FS22-053, FS22-053-008, FS22-053-009, FS22-053-010, FS22-053-011, FS22-053-012, FS22-053-013, FS22-053-014, FS22-053-015, FS22-053-016, FS22-053-017, FS22-172, FS22-172-001, FS22-172-002, FS22-172-003, FS22-172-004, FS22-172-005 and FS22-172-006 is shown in SEQ ID NO: 175, 21, 30, 39, 49, 58, 66, 75, 84, 93, 102, 112, 121, 130, 139, 148, 157 and 166, respectively.

The first and second sequence of the specific binding element FS22-053, FS22-053-008, FS22-053-009, FS22-053-010, FS22-053-011, FS22-053-012, FS22-053-013, FS22-053-014, FS22-053-015, FS22-053-016, FS22-053-017, FS22-172, FS22-172-001, FS22-172-002, FS22-172-003, FS22-172-004, FS22-172-005 and FS22-172-006 may be a sequence between positions 14 and 17 and positions 91 and 99 of the CH3 domain of the specific binding element FS22-053, FS22-053-008, FS22-053-009, FS22-053-010, FS22-053-011, FS22-053-012, FS22-053-013, FS22-053-014, FS22-053-015, FS22-053-016, FS22-053-017, FS22-172, FS22-172-001, FS22-172-002, FS22-172-003, FS22-172-004, FS22-172-005 and FS22-172-006, respectively.

Alternatively, the first and second sequence of the specific binding element FS22-053-008, FS22-053-010, FS22-053-011, FS22-053-012 and FS22-053-016 may be a sequence between positions 14 and 17 and positions 92 and 99 of the CH3 domain of the specific binding element FS22-053-008, FS22-053-010, FS22-053-011, FS22-053-012 and FS22-053-016, respectively.

The first and second sequence of the specific binding element FS22-053-015 may alternatively be a sequence between positions 14 and 17 and positions 92 and 98 of the CH3 domain of the specific binding element FS22-053-015, respectively.

The sequence of the CD loop of the specific binding element is preferably unmodified, i.e. wild type. Thus, the sequence of the CD loop preferably has the sequence presented in SEQ ID NO: 2. The sequence of the CD loop is preferably located at positions 43 to 78 of the CH3 domain of the specific binding element.

The first and second sequences may be the complete sequences of the structural loops AB and EF of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006, FS22-172, FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, respectively. Determining the location of the structural loops AB, CD and EF in the sequence of the CH3 domain, for example according to the IMGT, IMGT exon, EC or Kabat numbering systems, is within the capabilities of the skilled person and is described in Hasenhindl et al. (2013). According to a preferred embodiment, the structural loops AB, CD and EF according to the IMGT numbering system are located between positions 10 and 19, 42 and 79 and 91 and 102 of the CH3 domain of the specific binding element, respectively. Thus, according to a preferred embodiment, the first, second and third sequences are the sequence between positions 10 and 19, 42 and 79, and 91 and 102 of the CH3 domain of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016, FS22-053, FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, respectively.

According to a preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, specific binding element sequence:

(i) FS22-172-003, shown in SEQ ID NO: 141;

(ii) FS22-172-002, shown in SEQ ID NO: 132;

(iii) FS22-172-004, shown in SEQ ID NO: 150;

(iv) FS22-172-001, shown in SEQ ID NO: 123;

(v) FS22-172-005, shown in SEQ ID NO: 159;

(vi) FS22-172-006, as represented by SEQ ID NO: 167; or

(vii) FS22-172 as set forth in SEQ ID NO: 114; wherein said first and second sequences are preferably located between positions 14 and 17, and 91 and 99 of the CH3 domain of the specific binding element, respectively.

According to a more preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, specific binding element sequence:

(i) FS22-172-003, shown in SEQ ID NO: 141;

(ii) FS22-172-002, shown in SEQ ID NO: 132;

(iii) FS22-172-004, shown in SEQ ID NO: 150;

(iv) FS22-172-001, shown in SEQ ID NO: 123;

(v) FS22-172-005, as shown in SEQ ID NO: 159; or

(vi) FS22-172-006, represented by SEQ ID NO: 167.

According to another preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, specific binding element sequence:

(i) FS22-172-003, shown in SEQ ID NO: 141;

(ii) FS22-172-002, as shown in SEQ ID NO: 132; or

(iii) FS22-172-004, shown in SEQ ID NO: 150.

According to an even more preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, sequence of the specific binding element FS22-172-003, presented in SEQ ID NO: 141.

According to a preferred embodiment, the specific binding element, in particular the specific binding element comprising the first and/or second, preferably first and second, sequence of the specific binding element FS22-172-006, may comprise a leucine (L) at position 19 of the CH3 domain of the specific binding element.

According to an alternative preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, specific binding element sequence:

(i) FS22-053-008, shown in SEQ ID NO: 23;

(ii) FS22-053-009, shown in SEQ ID NO: 32;

(iii) FS22-053-011, represented by SEQ ID NO: 50;

(iv) FS22-053-017, shown in SEQ ID NO: 104;

(v) FS22-053-014, shown in SEQ ID NO: 77;

(vi) FS22-053-010, shown in SEQ ID NO: 41;

(vii) FS22-053-012, shown in SEQ ID NO: 59;

(viii) FS22-053-013, shown in SEQ ID NO: 68;

(ix) FS22-053-015, represented by SEQ ID NO: 86;

(x) FS22-053-016, as represented by SEQ ID NO: 95; or

(xi) FS22-053, as set forth in SEQ ID NO: 15; wherein said first and second sequences are preferably located between positions 14 and 17, and 91 and 99 of the CH3 domain of the specific binding element, respectively.

According to a more preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, specific binding element sequence:

(i) FS22-053-008, shown in SEQ ID NO: 23;

(ii) FS22-053-009, shown in SEQ ID NO: 32;

(iii) FS22-053-011, represented by SEQ ID NO: 50;

(iv) FS22-053-017, as represented by SEQ ID NO: 104; or

(v) FS22-053-014, shown in SEQ ID NO: 77.

According to an even more preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, CH3 domain sequence of the specific binding element:

(i) FS22-053-008, shown in SEQ ID NO: 23;

(ii) FS22-053-009, shown in SEQ ID NO: 32;

(iii) FS22-053-011, as represented by SEQ ID NO: 50; or

(iv) FS22-053-017, represented by SEQ ID NO: 104.

According to an even more preferred embodiment, the specific binding element comprises a first and/or second, preferably first and second, sequence of the specific binding element FS22-053-008, presented in SEQ ID NO: 23.

As an alternative to IMGT numbering, amino acid residue positions, including the positions of amino acid sequences, substitutions, deletions and insertions described in this application, may be numbered according to IMGT exon numbering (also referred to as sequential numbering), EU numbering, or Kabat numbering. The correspondence between the IMGT numbering, the IMGT exon numbering, the EC numbering and the Kabat numbering of the CH3 domain residue positions is shown in Figure 1. Thus, for example, where the present application relates to a first sequence located between positions 14 and 17 of the CH3 domain of a specific binding element, respectively, where the residue positions are numbered according to the IMGT numbering scheme, the first sequence is located between positions 18 and 21 of the CH3 domain, wherein the residue positions are numbered according to the IMGT exon numbering scheme as shown in Figure 1. Alternatively, the positions of amino acid residues in the CH3 domain, including the positions of the amino acid sequences, substitutions, deletions and insertions in the CH3 domain described herein, may be determined by reference to their position in the wild-type CH3 domain sequence set forth in SEQ ID NO: 4. The correspondence between IMGT numbering and wild-type CH3 domain sequence are also shown in Figure 1.

According to a preferred embodiment, the specific binding member comprises a CH3 domain that comprises, has or consists of the sequence of the CH3 domain of the specific binding member FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably the sequence of the CH3 domain of the specific binding member FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, more preferably the sequence of the CH3 domain of the specific binding member FS22-172-003, FS22-172-002 or FS22-172-004, even more preferably the CH3 domain sequence of the specific binding element FS22-172-003.

According to an alternative preferred embodiment, the specific binding member comprises a CH3 domain that comprises, has or consists of the CH3 domain sequence of the specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably the CH3 domain sequence of the specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably the CH3 domain sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably the CH3 domain sequence of the specific binding element FS22-053-008.

The CH3 domain of the specific binding element may optionally contain an additional lysine (K) residue immediately at the C-terminus of the CH3 domain sequence.

To produce other antibodies or chimeric molecules that retain the specificity of the original antibody, monoclonal and other antibodies and recombinant DNA technology techniques can be used. Such techniques can involve the introduction of CDRs or variable regions into another immunoglobulin. The introduction of CDRs from one immunoglobulin into another immunoglobulin is described, for example, in EP-A-184187, GB 2188638A and EP-A-239400. Similar techniques can be used to introduce constant domain sequences constituting the antigen-binding site for CD137 of the specific binding member of the present invention into a constant domain, for example the CH3 domain, of another specific binding member, thereby producing a specific binding member comprising an antigen-binding site for CD137 in its constant domain. Alternatively, the entire constant domain sequence of the specific binding member may be replaced with the constant domain sequence of the specific binding member of the present invention to obtain a specific binding member comprising an antigen-binding site for CD137 in its constant domain. Similarly, a fragment of the constant domain sequence of the specific binding member may be replaced with a corresponding fragment of the constant domain sequence of the specific binding member of the present invention comprising an antigen-binding site for CD137.

The CH2 domain of the specific binding element may comprise one or more mutations that reduce or eliminate binding of the CH2 domain to one or more Fcγ receptors, such as FcγRI, FcγRIIa, FcγRIIb, FcγRIII, and/or complement. The present inventors contemplate that reducing or eliminating binding to Fcγ receptors will reduce or eliminate ADCC mediated by the specific binding element. Similarly, reducing or eliminating binding to complement is expected to reduce or eliminate complement-dependent cytotoxicity (CDC) mediated by the specific binding element. Mutations to reduce or eliminate binding of the CH2 domain to one or more Fcγ receptors and/or complement are known in the art (Wang et al., 2018). These mutations include the “LALA mutation” described in Bruhns et al., 2009 and Hezareh et al., 2001, which involves replacing the leucine residues at positions 1.3 and 1.2 of the IMGT CH2 domain with alanine (L1.3A and L1.2A). Alternatively, production of a-glycosyl antibodies by mutation of the conserved N-linked glycosylation site by mutating the aparagine (N) at position 84.4 of the IMGT CH2 domain to alanine, glycine, or glutamine (N84.4A, N84.4G, or N84.4Q) also reduces IgG1 effector function (Wang et al., 2018). As another alternative, complement activation (C1q binding) and ADCC are known to be reduced by mutation of proline at position 114 of the IMGT CH2 domain to alanine or glycine (P114A or P114G) (Idusogie et al., 2000; Klein et al., 2016). These mutations can also be combined to generate specific binding elements with or without further reduced ADCC or CDC activity.

Thus, the specific binding element may comprise a CH2 domain, wherein said CH2 domain preferably comprises:

(i) alanine residues at positions 1.3 and 1.2; and/or

(ii) alanine or glycine at position 114; and/or

(iii) alanine, glutamine or glycine at position 84.4;

Moreover, the indicated numbering of amino acid residues corresponds to the IMGT numbering scheme.

According to a preferred embodiment, the specific binding element comprises a CH2 domain, wherein said CH2 domain preferably comprises:

(i) alanine residues at positions 1.3 and 1.2; and/or

(ii) alanine or glycine at position 114;

Moreover, the indicated numbering of amino acid residues corresponds to the IMGT numbering scheme.

According to another preferred embodiment, the specific binding element comprises a CH2 domain, wherein said CH2 domain comprises:

(i) an alanine residue at position 1.3; and

(ii) an alanine residue at position 1.2;

Moreover, the indicated numbering of amino acid residues corresponds to the IMGT numbering scheme.

For example, the CH2 domain may have the sequence shown in SEQ ID NO: 6. [LALA]

According to an alternative preferred embodiment, the specific binding element comprises a CH2 domain, wherein said CH2 domain comprises:

(i) alanine residue at position 1.3;

(ii) an alanine residue at position 1.2; and

(iii) alanine at position 114;

Moreover, the indicated numbering of amino acid residues corresponds to the IMGT numbering scheme.

For example, the CH2 domain may have the sequence shown in SEQ ID NO: 5. [LALA-PA]

According to a preferred embodiment, the specific binding member comprises, has or consists of: the CH2 and CH3 domain sequence of the specific binding member FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably the CH2 and CH3 domain sequence of the specific binding member FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, more preferably the CH2 and CH3 domain sequence of the specific binding member FS22-172-003, FS22-172-002 or FS22-172-004, even more preferably the CH2 and CH3 domain sequence of the specific binding member of FS22-172-003, wherein said CH2 and CH3 domain sequence of the specific binding member of FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006, and FS22-172 is shown in SEQ ID NOs 141, 132, 150, 123, 159, 167, and 114, respectively, starting from amino acid 7 onwards.

According to an alternative preferred embodiment, the specific binding member comprises, has or consists of: the CH2 and CH3 domain sequence of the specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably the CH2 and CH3 domain sequence of the specific binding member FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably the CH2 and CH3 domain sequences of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably the CH2 and CH3 domain sequences of the specific binding element FS22-053-008, wherein said sequence of the CH2 and CH3 domain of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 and FS22-053 are shown in SEQ ID NOs 23, 32, 50, 104, 77, 41, 59, 68, 86, 95 and 15, respectively, from amino acid 7 onwards.

According to an alternative preferred embodiment, the specific binding element comprises, has or consists of the specific binding element sequence FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably the specific binding element sequence FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably the sequence specific binding element FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably the sequence of the specific binding element FS22-053-008, wherein said sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 and FS22-053 is given in SEQ ID NO 23, 32, 50, 104, 77, 41, 59, 68, 86, 95 and 15, respectively.

According to a preferred embodiment, the specific binding member may comprise one or more additional antigen-binding sites that bind one or more additional antigens, in addition to the antigen-binding site for CD137 located in the constant domain of the specific binding member. The one or more additional antigen-binding sites preferably specifically bind their cognate antigens.

One or more additional antigen-binding sites may bind CD137 or another antigen. Thus, the specific binding member may be a multispecific, such as a bispecific, trispecific or tetraspecific molecule, preferably a bispecific molecule. According to a preferred embodiment, the specific binding member is capable of simultaneously binding to CD137 and one or more additional antigens.

Antibody molecules are known to have a modular architecture comprising discrete domains that can be combined in a variety of ways to form multispecific, e.g. bispecific, trispecific or tetraspecific antibody formats. Exemplary multispecific antibody formats are described in Spiess et al. (2015) and Kontermann (2012), by way of example. The specific binding members of the present invention can be used in such multispecific antibody formats. This has the additional advantage of introducing an additional antigen-binding site into such a multispecific antibody format by having an antigen-binding site of a constant domain, e.g. a CH3 domain, of the specific binding member.

For example, a specific binding member of the present invention may be a heterodimeric antibody molecule, such as a complete heterodimeric immunoglobulin molecule, or a fragment thereof. In this case, one portion of the antibody molecule will have the sequence or sequences described herein. For example, if a specific binding member of the present invention is a bispecific heterodimeric antibody molecule, the specific binding member may comprise a heavy chain comprising a CH3 domain described herein paired with a heavy chain that binds an antigen other than CD137. Techniques for producing heterodimeric antibodies are known in the art and include knob-in-hole (KIH) technology, which involves engineering the CH3 domains of an antibody molecule to create both a knob and a hole to promote heterodimerization of the chains. Alternatively, heterodimeric antibodies can be produced by introducing charge pairs into the antibody molecule to avoid homodimerization of the CH3 domains by electrostatic repulsion and to direct heterodimerization by electrostatic attraction. Examples of heterodimeric antibody formats include CrossMab, mAb-Fv, SEED-body, and KIH IgG.

Alternatively, the multispecific specific binding member of the present invention may comprise a complete immunoglobulin molecule or a fragment thereof and an additional antigen-binding group or groups. The antigen-binding group may be, for example, an Fv, scFv or a single domain antibody and may be fused to a complete immunoglobulin molecule or a fragment thereof. Examples of multispecific antibody molecules comprising additional antigen-binding groups fused to a complete immunoglobulin molecule include DVD-IgG, DVI-IgG, scFv4-IgG, IgG-scFv and scFv-IgG molecules (Spiess et al., 2015; Figure 1). Examples of multispecific antibody molecules containing additional antigen-binding moieties fused to a CH3 domain-containing immunoglobulin fragment include, for example, scDiabody-CH3, Diabody-CH3, and scFv-CH3 KIH (Spiess et al., 2015; Figure 1).

Other suitable multi-specific formats will be easily understood by a qualified professional.

According to a preferred embodiment, the specific binding member comprises a second antigen-binding site that binds a second antigen, wherein said second antigen-binding site is preferably a CDR-based antigen-binding site. The CDR-based antigen-binding site is an antigen-binding site in the variable region of the antibody. The CDR-based antigen-binding site is formed by six CDRs; three CDRs of the variable domain of the light chain (VL) and three CDRs of the variable domain of the heavy chain (VH).

The production of antibody molecules against a specific antigen and the determination of the CDR sequences of such antibody molecules are well known and many suitable techniques are known in the art. The CDR sequences can be determined, for example, according to Kabat et al., 1991 or the international ImMunoGeneTics (IMGT) information system (Lefranc et al., 2015).

For example, the specific binding member may be a bispecific antibody MAB ² (TM). The bispecific antibody MAB ² referred to in the present application is an IgG immunoglobulin that comprises a CDR-based antigen-binding site in each of its variable regions and at least one antigen-binding site in the constant domain. If the specific binding member of the present invention is in the MAB ² format, the specific binding member accordingly comprises a CDR-based antigen-binding site in each of its variable regions in addition to an antigen-binding site for CD137 in the constant domain of the specific binding member.

The three CDRs of the VH domain of the antigen-binding site may be located in the VH domain of the immunoglobulin, and the three CDRs of the VL domain may be located in the VL domain of the immunoglobulin. For example, the CDR-based antigen-binding site may be located in the variable region of the antibody.

The specific binding member may have one or preferably more than one, such as two, CDR-based antigen-binding sites for the second antigen. Thus, the specific binding member may comprise one VH domain and one VL domain, but preferably comprises two VH domains and two VL domains, i.e. two VH/VL domain pairs, as in the case of natural IgG molecules.

According to some preferred embodiments, the specific binding member may be an immunoglobulin comprising two variable regions, each variable region comprising a CDR-based antigen-binding site for a second antigen.

Thus, according to a preferred embodiment, the antibody is an antibody molecule that binds CD137 and a second antigen, wherein said antibody molecule comprises:

(i) two antigen-binding sites for CD137, which are located in two CH3 domains of the antibody molecule; and

(ii) two CDR-based antigen-binding sites for a second antigen, each formed by an immunoglobulin VH domain and an immunoglobulin VL domain.

According to a more preferred embodiment, the antibody is a complete immunoglobulin molecule, such as a complete IgG1 molecule, that binds CD137 and a second antigen, wherein said antibody molecule comprises:

(i) two antigen-binding sites for CD137, which are located in two CH3 domains of the antibody molecule; and

(ii) two CDR-based antigen-binding sites for a second antigen, each formed by an immunoglobulin VH domain and an immunoglobulin VL domain; and

wherein the said immunoglobulin molecule additionally contains CH1, CH2 and CL domains.

CD137 activation requires clustering of CD137 on the surface of an immune cell, such as a T cell, which in turn stimulates intracellular signaling pathways and immune cell activation. Binding of specific binding elements to CD137 on the surface of an immune cell in the absence of cross-linking of specific binding elements may not induce clustering of CD137 and therefore may not result in immune cell activation.

The present inventors have shown that the specific binding elements of the FS22-53 and FS22-172 lines do not induce T cell activation in the absence of cross-linking of the specific binding element (see Example 5).

As explained above, cross-linking of antibody molecules via binding to Fcγ receptors is inefficient and cannot be targeted to a specific site, such as a disease site, since cells expressing the Fcγ receptor are present throughout the human body. Thus, the second antigen bound by the second antigen-binding site is preferably not an Fcγ receptor.

Thus, according to a preferred embodiment, the specific binding members of the present invention comprise a second antigen-binding site that binds a second antigen, wherein said second antigen is capable of binding and cross-linking to a plurality of specific binding members.

For example, the present inventors have shown that if the second antigen is a multimeric molecule, binding of a specific binding element to the second antigen results in or enhances T cell activation. Thus, the second antigen is preferably a multimeric antigen, such as a dimer, trimer or higher order multimer, and is accordingly capable of cross-linking several specific binding elements.

The present inventors have also shown, using anti-CD137/second antigen MAb ² molecules, that when the second antigen is a surface antigen, such as a cell surface antigen, which may be monomeric or multimeric and is present in high concentrations and/or clustered on a surface, such as a cell surface, binding of an antibody molecule to the second antigen results in or enhances T cell activation. Without being limited by theory, it is believed that binding of an antibody molecule to a common cell surface antigen, such as a cell surface, results in a high concentration of antibody molecules bound to the cell surface, which ensures that the antibody molecules are placed in sufficient proximity to be able to trigger CD137 clustering and immune cell activation. Thus, in a preferred embodiment, the second antigen is a surface antigen that is expressed in high concentration on a surface, such as a cell surface.

A specific binding member comprising a second antigen-binding site that binds a second antigen as described herein and that activates immune cells, such as T cells, only upon binding to the second antigen or whose immune cell activating activity is enhanced upon binding to the second antigen is also referred to as a conditional agonist. The immune cell activating activity upon binding to the second antigen is independent of the binding of the specific binding member to Fcγ receptors and/or external cross-linking agents such as protein A or G or secondary antibodies and, therefore, allows the conditional agonist activity of the specific binding member to be targeted to sites where the second antigen is present. For example, if the second antigen is a disease antigen, the specific binding member may selectively activate an immune cell at the site of the disease and not elsewhere in the individual.

In addition, a specific binding member that activates immune cells, such as T cells, only upon binding to a second antigen preferably has increased activity in activating an immune cell compared to specific binding members that depend on cross-linking by other mechanisms, such as external cross-linking agents or cross-linking by interaction with an Fcγ receptor. Since CD137 activation is more efficient, immune cell activation can be achieved at lower concentrations of the specific binding members described herein compared to other specific binding members.

Thus, the specific binding element of the present invention preferably induces increased activation of immune cells, such as T cells, when the specific binding element is cross-linked, for example by binding to a second antigen, than in the absence of cross-linking of the specific binding element.

The ability of an antibody molecule or specific binding element to activate T cells can be measured using a T cell activation assay. When activated, T cells release IL-2. Thus, a T cell activation assay can measure the release of IL-2 to determine the level of T cell activation induced by an antibody molecule or specific binding element.

For example, the ability of an antibody molecule or specific binding member to activate T cells can be determined by measuring the concentration of the antibody molecule or specific binding member required to achieve half-maximal IL-2 release by T cells in a T cell activation assay when the specific binding member or antibody molecule is cross-linked. This is referred to hereinafter as the EC ₅₀ of the antibody molecule or specific binding member. A lower EC ₅₀ indicates that a lower concentration of the antibody molecule or specific binding member is required to achieve half-maximal IL-2 release by T cells in a T cell activation assay and, accordingly, that the antibody molecule or specific binding member has higher activity in activating T cells. The specific binding member or antibody molecule can be cross-linked using an antibody to CH2, as an example.

According to a preferred embodiment, the antibody molecule or specific binding member has an _EC50 in a T cell activation assay that is within 10-fold, 5-fold, 4-fold, 3-fold or 2-fold of the _EC50 of FS22-172-003/HelD1.3 (containing the LALA mutation) in the same assay, wherein FS22-172-003/HelD1.3 (LALA) consists of or comprises the heavy chain shown in SEQ ID NO: 145 and the light chain shown in SEQ ID NO: 173.

According to an alternative preferred embodiment, the antibody molecule or specific binding member has an _EC50 in a T cell activation assay that is within 10-fold, 5-fold, 4-fold, 3-fold or 2-fold of the _{EC50 of} FS22-053-008/HelD1.3 (containing the LALA mutation) in the same assay, wherein FS22-053-008/HelD1.3 (LALA) consists of or comprises the heavy chain shown in SEQ ID NO: 27 and the light chain shown in SEQ ID NO: 173.

For example, an antibody molecule or specific binding member may have an _EC50 in a T cell activation assay of 5 nM or less, 4 nM or less, 3 nM or less, 2 nM or less, 1 nM or less, or 0.5 nM or less.

Additionally or alternatively, the ability of an antibody molecule or specific binding member to activate T cells can be determined by measuring the maximum concentration of IL-2 released by T cells in a T cell activation assay in the presence of the antibody molecule or specific binding member, wherein said antibody molecule or specific binding member is cross-linked.

According to a preferred embodiment, the maximum concentration of IL-2 released by T cells in a T cell activation assay in the presence of an antibody molecule or a specific binding member in the presence of cross-linking is within 3-fold, 2-fold or 1.5-fold the maximum concentration of IL-2 released by T cells in the presence of FS22-053-008/HelD1.3 (containing the LALA mutation) or FS22-172-003/HelD1.3 (containing the LALA mutation).

The T cell activation assay may be a T cell assay described herein, such as the CD8 ⁺ T cell assay described herein in the Examples, see, for example, Example 5.4.

For example, the T cell activation assay may be an IL-2 release assay based on CD8 ⁺ T cells isolated from human peripheral blood mononuclear cells (PBMCs). For example, the T cell activation assay may include isolating human PBMCs from leukocyte depletion cones. Methods for isolating PBMCs are known in the art and are described in the examples of the present application. CD8 ⁺ T cells may then be isolated from the PBMCs. Methods for isolating CD8 ⁺ T cells from PBMCs are known in the art and are described in the examples of the present application.

The CD8 ⁺ T cells can then be added to multi-well plates coated with an anti-human CD3 antibody. An appropriate dilution of each antibody molecule or specific binding member to be tested can be prepared and added to the wells. The T cells can then be incubated at 37°C, 5% _CO2 for 24 hours with the antibody to be tested. The supernatants can be collected and analyzed to determine the concentration of IL-2 in the supernatant. Methods for determining the concentration of IL-2 in solution are known in the art and are described in the examples of the present application. The concentration of human IL-2 can be plotted against the logarithm of the concentration of the antibody molecule or specific binding member. The resulting curves can be approximated using the Log(agonist) response equation.

The second antigen bound by the second antigen-binding site of the specific binding member may be an immune cell antigen or a disease antigen. Disease antigens include pathogen antigens and tumor antigens.

The immune cell antigen bound by the specific binding element may be present on the same immune cell or on a different immune cell than CD137.

The immune cell antigen may be a member of the tumor necrosis factor receptor superfamily (TNFRSF) other than CD137. TNFRSF receptors are membrane-bound cytokine receptors that contain an extracellular cysteine-rich domain that binds one or more tumor necrosis factor superfamily (TNFSF) ligands.

The TNFRSF receptor can be located on the surface of an immune cell. Upon binding of a TNFRSF ligand, TNFRSF receptors form clusters on the surface of the immune cell that activate the immune cell. For example, ligand-bound TNFRSF receptors can form multimers, such as trimers, or clusters of multimers. The presence of clusters of ligand-bound TNFRSF receptors stimulates intracellular signaling pathways that activate the immune cell.

Without being bound by theory, it is believed that by binding to both CD137 and a second TNFRSF receptor on the surface of an immune cell, the specific binding elements will induce both CD137 and the second TNFRSF receptor to cluster and activate the immune cell(s). In other words, the specific binding element will act as a TNFRSF receptor agonist when both targets are bound.

TNFRSF receptors include CD27, CD40, EDA2R, EDAR, FAS, LTBR, RELT, TNFRSF1A, TNFRSF1B, TNFRSF4, TNFRSF6B, TNFRSF8, TNFRSF10A-10D, TNFRSF11A, TNFRSF11B, TNFRSF12A, TNFRSF13B, TNFRSF13C, TNFRSF14, TNFRSF17, TNFRSF18, TNFRSF19, TNFRSF21, and TNFRSF25.

CD27 (TNFRSF7: Gene ID 939) has the reference amino acid sequence NP_001233.1 and can be encoded by the reference nucleotide sequence NM_001242.4. CD40 (TNFRSF5: Gene ID 958) has the reference amino acid sequence NP_001241.1 and can be encoded by the reference nucleotide sequence NM_001250.5. EDA2R (TNFRSF27: Gene ID 60401) has the reference amino acid sequence NP_001186616.1 and can be encoded by the reference nucleotide sequence NM_001199687.2. EDAR (Gene ID 10913) has the reference amino acid sequence NP_071731.1 and can be encoded by the reference nucleotide sequence NM_022336, 3. FAS (TNFRSF6: Gene ID 355) has the reference amino acid sequence NP_000034.1 and can be encoded by the reference nucleotide sequence NM_000043.5. LTBR (TNFRSF3: Gene ID 4055) has the reference amino acid sequence NP_001257916.1 and can be encoded by the reference nucleotide sequence NM_001270987.1. RELT (TNFRSF19L: Gene ID 84957) has the reference amino acid sequence NP_116260.2 and can be encoded by the reference nucleotide sequence NM_032871.3. TNFRSF1A (Gene ID 7132) has the reference amino acid sequence NP_001056.1 and can be encoded by the reference nucleotide sequence NM_001065.3. TNFRSF1B (Gene ID 7133) has the reference amino acid sequence NP_001057.1 and can be encoded by the reference nucleotide sequence NM_001066.2. TNFRSF4 (Gene ID 7293) has the reference amino acid sequence NP_003318 and can be encoded by the reference nucleotide sequence NM_003327). TNFRSF6B (Gene ID 8771) has the reference amino acid sequence NP_003814.1 and can be encoded by the reference nucleotide sequence NM_003823.3. TNFRSF8 (Gene ID 943) has the reference amino acid sequence NP_001234.3 and can be encoded by the reference nucleotide sequence NM_001243.4. TNFRSF10A (Gene ID 8797) has the reference amino acid sequence NP_003835.3 and can be encoded by the reference nucleotide sequence NM_003844.3. TNFRSF10B (Gene ID 8795) has the reference amino acid sequence NP_003833.4 and can be encoded by the reference nucleotide sequence NM_003842.4. TNFRSF10C (Gene ID 8794) has the reference amino acid sequence NP_003832.2 and can be encoded by the reference nucleotide sequence NM_003841.4. TNFRSF10D (Gene ID 8793) has the reference amino acid sequence NP_003831.2 and can be encoded by the reference nucleotide sequence NM_003840.4. TNFRSF11A (Gene ID 8792) has the reference amino acid sequence XP_011524547.1 and can be encoded by the reference nucleotide sequence XM_11526245.2. TNFRSF11B (Gene ID 4982) has the reference amino acid sequence NP_002537.3 and can be encoded by the reference nucleotide sequence NM_002546.3. TNFRSF12A (Gene ID 51330) has the reference amino acid sequence NP_057723.1 and can be encoded by the reference nucleotide sequence NM_016639.2. TNFRSF13B (Gene ID 23495) has the reference amino acid sequence NP_0036584.1 and can be encoded by the reference nucleotide sequence NM_012452.2. TNFRSF13C (Gene ID 115650) has the reference amino acid sequence NP_443177.1 and can be encoded by the reference nucleotide sequence NM_052945.3. TNFRSF14 (Gene ID 8764) has the reference amino acid sequence NP_001284534.1 and can be encoded by the reference nucleotide sequence NM_001297605.1. TNFRSF17 (Gene ID 608) has the reference amino acid sequence NP_001183.2 and can be encoded by the reference nucleotide sequence NM_001192.2. TNFRSF18 (Gene ID 8784) has the reference amino acid sequence NP_004195.2 and can be encoded by the reference nucleotide sequence NM_004186.1. TNFRSF19 (Gene ID 55504) has the reference amino acid sequence NP_001191387.1 and can be encoded by the reference nucleotide sequence NM_001204458.1. NFRSF21 (Gene ID 27242) has the reference amino acid sequence NP_055267.1 and can be encoded by the reference nucleotide sequence NM_014452.4. TNFRSF25 (DR3: gene identifier 8718) binds to the TNFSF15 ligand (TL1A), has the reference amino acid sequence NP_001034753.1 and may be encoded by the reference nucleotide sequence NM_001039664.1.

Alternatively, the immune cell antigen bound by the second antigen-binding site may be a molecule that has a regulatory function in the immune system other than a TNFRSF member, such as an immune costimulatory molecule or a checkpoint inhibitor molecule. Examples of such immune regulatory molecules include ICOS (CD278), LAG3, PD1, PD-L1, PD-L2, B7H3, B7H4, CTLA4, TIGIT, BTLA, HVEM, mucin domain-containing T cell immunoglobulin 3 (TIM-3), CD47, CD73, A2aR, CD200, CD200R, colony stimulating factor receptor 1 (CSF-1R), VISTA CD28, CD80, LLT1, galectin-9, NKG2A, NKG2D and KIR.

The immune cell on which the immune cell antigen is present may belong to any subset of immune cells and may be a T cell, a tumor infiltrating leukocyte (TIL), a cell of the myeloid lineage such as an antigen-presenting cell (APC), a NK cell, and/or a B cell. In the case where the immune cell antigen is a TNFRSF receptor, the immune cell on which the TNFRSF receptor is present is preferably a T cell.

Alternatively, the second antigen-binding site may bind to a disease antigen as mentioned above. Without being bound by theory, it is believed that binding of the specific binding element to CD137 and the disease antigen will result in activation of T cells in the vicinity of the disease. The activated T cells can then initiate, promote, or participate in an immune response, such as an immune response against a pathogen or a cancer cell. The role of the immune system in recognizing and eliminating cancer cells is reviewed by Chen and Mellman (2013).

According to a preferred embodiment, the disease antigen is a tumor antigen. A tumor antigen is an antigen that is predominantly present in the tumor environment and is not ubiquitously present elsewhere in the individual. For example, a tumor antigen may be present on the surface of tumor cells or may be present on other stromal cells in the tumor microenvironment or in biological fluids near the tumor. Thus, a tumor antigen is a marker of the location of tumor cells in an individual.

In some embodiments, the tumor antigen may be an antigen that is located on the surface of a cancer cell. Preferably, the tumor antigen is up-regulated or overexpressed on tumor cells, while it is not expressed in large quantities by corresponding normal somatic cells from the same tissue in the absence of a tumor.

In some embodiments, the tumor antigen is upregulated or overexpressed on stromal cells of the tumor microenvironment compared to stromal cells of the corresponding normal tissue in the absence of the tumor.

Preferred tumor antigens exist on the cell surface and are not rapidly internalized.

Tumor antigens that are suitable for targeting by specific binding members can be identified using methods known in the art. For example, a specific binding member targeting a CD137 receptor and a tumor antigen can be used in an assay in which a CD137-expressing cell is co-cultured with a cell expressing a tumor antigen and the activation of the CD137-expressing cell is measured, for example, by a T-cell activation assay, a proliferation assay, or a cytotoxicity assay.

A tumor cell surface antigen may be a tumor-associated antigen (TAA) or a tumor-specific antigen (TSA).

Tumor antigens expressed by cancer cells may include, for example, cancer-testicular antigens (CT) encoded by cancer germline genes such as MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, GAGE-I, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, BAGE-I, RAGE- 1, LB33/MUM-1, PRAME, NAG, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE- C1/CT7, MAGE-C2, NY-ESO-I, LAGE-I, SSX-I, SSX-2(HOM-MEL-40), SSX-3, SSX-4, SSX-5, SCP-I and XAGE and immunogenic fragments or variants thereof (Simpson et al., 2005; Gure et al., 2005; Velazquez et al., 2007; Andrade et al., 2008; Tinguely et al., 2008; Napoletano et al., 2008).

Other tumor cell surface antigens include, for example, AFP, α _v β ₃ (vitronectin receptor), α _v β ₆ , B cell maturation agent (BCMA), CA125 (MUC16), CD4, CD20, CD22, CD33, CD52, CD56, CD66e, CD80, CD140b, CD227 (MUC1), EGFR (HER1), EpCAM, ganglioside GD3, HER2, prostate-specific membrane antigen (PSMA), prostate-specific antigen (PSA), CD5, CD19, CD21, CD25, CD37, CD30, CD33, CD45, HLA-DR, anti-idiotype, carcinoembryonic antigen (CEA), such as carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), TAG-72, folate binding protein, A33, G250, ferritin, glycolipids such as gangliosides, carbohydrates such as CA-125, IL-2 receptor, fibroblast activation protein (FAP), IGF1R, B7H3, B7H4, PD-L1, CD200, EphA2, and mesothelin or their variants. These and other tumor cell surface antigens are described in Carter et al., 2004; Scott and Renner, 2001; and Cheever et al., 2009; Tai and Anderson, 2015; and Podojil and Miller, 2017.

Other tumor antigens include complexes of MHC and out-of-frame peptide formed through AUG-free translation initiation mechanisms used by stressed cancer cells (Malarkannan et al., 1999).

Other tumor antigens include peptide-MHC complexes on the surface of tumor cells or tumor microenvironment cells, wherein said peptide-MHC complexes comprise a tumor-specific neoantigen peptide fragment of a mutated intracellular tumor antigen, and wherein said peptide neoantigen carries one or more tumor-specific mutations (Gubin et al., 2015). Other tumor antigens are well known in the art (see, for example, WO00/20581; Cancer Vaccines and Immunotherapy (2000) Eds Stern, Beverley and Carroll, Cambridge University Press, Cambridge). The sequences of these tumor antigens are readily available from publicly available databases and can also be found in WO1992/020356 A1, WO1994/005304 A1, WO1994/023031 A1, WO1995/020974 A1, WO1995/023874 A1 and WO1996/026214 A1.

Preferred tumor antigens include HER2, FAP, EpCAM, CEACAM5, CD20, CD73, PSMA, mesothelin, EphA2, IGF1R, CD200, α _v β ₆ , BCMA, PD-L1, B7H3, B7H4, and EGFR.

According to a more preferred embodiment, the tumor antigen is mesothelin (MSLN).

According to an alternative more preferred embodiment, the tumor antigen is PD-L1.

HER2 (ERBB2; Gene ID 2064) may have the reference amino acid sequence NP_001005862.1 and may be encoded by the reference nucleotide sequence NM_001005862.2. FAP (Gene ID 2191) may have the reference amino acid sequence NP_001278736.1 and may be encoded by the reference nucleotide sequence NM_001291807.1. EpCAM (Gene ID 4072) may have the reference amino acid sequence NP_002345.2 and may be encoded by the reference nucleotide sequence NM_002354.2.

CEACAM5 (Gene Identifier 1048) may have the reference amino acid sequence NP_001278413.1 and may be encoded by the reference nucleotide sequence NM_001291484.2. CD20 (MS4A1; Gene Identifier 931) may have the reference amino acid sequence NP_068769.2 and may be encoded by the reference nucleotide sequence NM_021950.3. CD73 (NT5E; Gene Identifier 4907) may have the reference amino acid sequence NP_001191742.1 and may be encoded by the reference nucleotide sequence NM_001204813.1. PSMA (FOLH1; Gene Identifier 2346) may have the reference amino acid sequence NP_001014986.1 and may be encoded by the reference nucleotide sequence NM_001014986.1. Mesothelin (MSLN; Gene Identifier 10232) may have the reference amino acid sequence NP_001170826.1 and may be encoded by the reference nucleotide sequence NM_001177355.2. EphA2 (Gene Identifier 1969) may have the reference amino acid sequence NP_001316019.1 and may be encoded by the reference nucleotide sequence NM_001329090.1. IGF1R (Gene Identifier 3480) may have the reference amino acid sequence NP_000866.1 and may be encoded by the reference nucleotide sequence NM_000875.4. CD200 (Gene Identifier 4345) may have the reference amino acid sequence NP_001004196.2 and may be encoded by the reference nucleotide sequence NM_001004196.3. α _v β ₆ is a heterodimer consisting of the integrin alpha V subunit and the integrin beta 6 subunit. The integrin alpha V subunit (ITGAV; Gene Identifier 3685) may have the reference amino acid sequence NP_001138471.1 and may be encoded by the reference nucleotide sequence NM_001144999.2. Integrin subunit beta 6 (ITGB6; Gene Identifier 3694) may have the reference amino acid sequence NP_000879.2 and may be encoded by the reference nucleotide sequence NM_000888.4. BCMA (TNFRSF17; Gene Identifier 608) may have the reference amino acid sequence NP_001183.2 and may be encoded by the reference nucleotide sequence NM_001192.2. PD-L1 (CD274; Gene Identifier 29126) may have the reference amino acid sequence NP_001254635.1 and may be encoded by the reference nucleotide sequence NM_001267706.1. B7H3 (CD276; Gene Identifier 80381) may have the reference amino acid sequence NP_001019907.1 and may be encoded by the reference nucleotide sequence NM_001024736.1. B7H4 (VTCN1; Gene Identifier 79679) may have the reference amino acid sequence NP_001240778.1 and may be encoded by the reference nucleotide sequence NM_001253849.1. EGFR (Gene Identifier 1956) may have the reference amino acid sequence NP_001333826.1 and may be encoded by the reference nucleotide sequence NM_001346897.1.

In other embodiments, the tumor antigen may be a soluble tumor antigen, such as a growth factor that is produced by or in response to cancer cells. The soluble factor may be upregulated or overexpressed in biological fluids in the vicinity of the tumor. The soluble tumor antigen may be multimeric, such as a dimer or trimer. The soluble tumor antigen may be present in higher concentrations at the tumor site or in the tumor microenvironment than elsewhere in the individual. The tumor microenvironment and associated soluble tumor antigens are described in more detail in Bhome et al. (2015).

Suitable soluble tumor antigens include VEGF, HGF, SDF1, and TGF-beta, such as TGF-beta-1, TGF-beta-2, TGF-beta-3, and TGF-beta-4.

VEGF (VEGFA; Gene Identifier 7422) has the reference amino acid sequence NP_001020537.2 and can be encoded by the reference nucleotide sequence NM_001025366.2. HGF (Gene Identifier 3082) has the reference amino acid sequence NP_000592.3 and can be encoded by the reference nucleotide sequence NM_000601.5. SDF1 (CXCL12; Gene Identifier 6387) has the reference amino acid sequence NP_000600.1 and can be encoded by the reference nucleotide sequence NM_000609.6. TGF-beta-1 (TGFB1; Gene Identifier 7040) may have the reference amino acid sequence NP_000651.3 and may be encoded by the reference nucleotide sequence NM_000660.6. TGF-beta-2 (TGFB2; Gene Identifier 7042) may have the reference amino acid sequence NP_001129071.1 and may be encoded by the reference nucleotide sequence NM_001135599.3. TGF-beta-3 (TGFB3; Gene Identifier 7043) may have the reference amino acid sequence NP_001316867.1 and may be encoded by the reference nucleotide sequence NM_001329938.1. TGF-beta-4 (LEFTY2; gene identifier 7044) may have the reference amino acid sequence NP_001165896.1 and may be encoded by the reference nucleotide sequence NM_001172425.2.

According to an alternative preferred embodiment, the disease antigen is a pathogen antigen.

It is expected that activation of immune cells such as T cells, NK cells and/or macrophages by a specific binding element in the immediate vicinity of an infectious disease site can be used in the treatment of an infectious disease. The infectious disease may be acute or persistent infectious diseases, but preferably persistent infectious diseases.

The pathogen antigen is preferably an antigen expressed by a human pathogen, such as a viral, bacterial, fungal or parasitic antigen (e.g., a protozoan antigen), preferably a viral or bacterial antigen. The pathogen antigen is an antigen that is predominantly present on the pathogen or in close proximity to the site of infectious disease and is not ubiquitously present elsewhere in the individual.

For example, the pathogen antigen may be an antigen present on the surface of a virus, bacterium, fungus, or parasite, or a soluble antigen expressed by a virus, bacterium, fungus, or parasite. The virus, bacterium, fungus, or parasite may be a virus, bacterium, fungus, or parasite mentioned elsewhere in this application.

If the pathogen antigen is a soluble antigen, the antigen may be upregulated or overexpressed in body fluids in the immediate vicinity of the infectious disease site. For example, a soluble pathogen antigen may be present in higher concentrations at or near the infectious disease site than elsewhere in the individual. A soluble pathogen antigen may be multimeric, such as a dimer or trimer.

Pathogen antigens that are suitable for targeting by a specific binding member can be identified using methods known in the art. For example, a specific binding member targeting CD137 and a pathogen antigen can be used in an assay in which a CD137-expressing cell is co-cultured with a pathogen or pathogen antigen, and the activation of the OX40-expressing cell is measured, for example, by a T-cell activation assay, a proliferation assay, or a cytotoxicity assay.

Many pathogen antigens suitable for targeting by a specific binding member are also known in the art and can be selected by a skilled person in accordance with the infectious disease to be treated. Examples of viral antigens include the proteins p24, gp120 and gp41 expressed by the human immunodeficiency virus (HIV), the hepatitis B surface antigen (HBsAg) expressed by the hepatitis B virus (HBV), and the hemagglutinin and neuraminidase expressed by the influenza virus. Examples of bacterial antigens include Rv1733, Rv2389 and Rv2435n expressed by Mycobacterium tuberculosis.

The specific binding member may also comprise a variant of the first or second sequence, the AB CD or EF structural loop sequence, the CH3 domain, the CH2 domain, the Fcab, the CDR, the VH domain, the VL domain, the light chain and/or the heavy chain sequence disclosed herein. Suitable variants may be obtained by sequence alteration or mutation and screening methods. According to a preferred embodiment, the specific binding member comprising one or more sequence variants retains one or more functional characteristics of the original specific binding member, such as binding specificity and/or binding affinity for CD137. For example, the specific binding member comprising one or more sequence variants preferably binds to CD137 with the same affinity or with a higher affinity than the (original) specific binding member. A parent specific binding member is a specific binding member that does not contain the amino acid substitution(s), deletion(s) and/or insertion(s) that was(were) included in the variant specific binding member.

For example, the specific binding member may comprise a first, second or third sequence, an AB, CD or EF structural loop sequence, a CH3 domain, a CH2 domain, an Fcab, a CDR, a VH domain, a VL domain, a light chain and/or a heavy chain sequence that has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity to the first, second or a third sequence, an AB, CD or EF structural loop sequence, a CH3 domain, a CH2 domain, Fcab, CDR, VH domain, VL domain, light chain or heavy chain sequence disclosed in this application.

According to a preferred embodiment, the specific binding member has or comprises a CH3 domain sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity, preferably at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity to the CH3 domain sequence shown in SEQ ID NO: 21, 30, 48, 102, 75, 39, 57, 66, 84, 93, 175, 139, 130, 148, 121, 157, 165 or 112.

According to another preferred embodiment, the specific binding member has or comprises a CH2 domain sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity to the CH2 domain sequence set forth in SEQ ID NO: 5 or 6.

According to another preferred embodiment, the specific binding member has, comprises or consists of a sequence that has at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity to the Fcab sequence shown in SEQ ID NO: 23, 32, 50, 104, 77, 41, 59, 68, 86, 95, 15, 141, 132, 150, 123, 159, 167, 114, 25, 34, 52, 106, 79, 43, 61, 70, 88, 97, 16, 143, 134, 152, 125, 161, 169 or 116.

Sequence identity is usually determined by reference to the GAP algorithm (Wisconsin GCG package, Accelerys Inc, San Diego, USA). GAP uses the Needleman and Wunsch algorithm to align two complete sequences, maximizing the number of matches and minimizing the number of gaps. Default parameters with a gap creation penalty of 12 and a gap expansion penalty of 4 are usually used. The use of GAP may be preferred, but other algorithms may be used, such as BLAST (which uses the method of Altschul et al. (1990)), FASTA (which uses the method of Pearson and Lipman (1988)), or the Smith–Waterman algorithm (Smith and Waterman (1981)), or the TBLASTN program of Altschul et al. (1990) above, usually using default parameters. In particular, the psi-Blast algorithm may be used.

The specific binding member may comprise a first, second or third sequence, an AB, CD or EF structural loop sequence, a CH3 domain, a CH2 domain, Fcab, CDR, VH domain, VL domain, light chain or heavy chain sequence that has one or more amino acid sequence changes (addition, deletion, substitution and/or insertion of an amino acid residue), preferably 20 changes or less, 15 changes or less, 10 changes or less, 5 changes or less, 4 changes or less, 3 changes or less, 2 changes or less or 1 change compared to the first, second or third sequence, an AB, CD or EF structural loop sequence, a CH3 domain, a CH2 domain, Fcab, CDR, VH domain, VL domain, light chain or heavy chain sequence disclosed in the present application.

According to a preferred embodiment, the specific binding member may comprise a CH3 domain sequence with one or more amino acid sequence changes (addition, deletion, substitution and/or insertion of an amino acid residue), preferably 20 changes or less, 15 changes or less, 10 changes or less, 5 changes or less, 4 changes or less, 3 changes or less, 2 changes or less, or 1 change compared to the CH3 domain sequence presented in SEQ ID NO: 21, 30, 48, 102, 75, 39, 57, 66, 84, 93, 175, 139, 130, 148, 121, 157, 165 or 112.

According to another preferred embodiment, the specific binding member comprises a CH2 domain sequence with one or more amino acid sequence changes (addition, deletion, substitution and/or insertion of an amino acid residue), preferably 20 changes or less, 15 changes or less, 10 changes or less, 5 changes or less, 4 changes or less, 3 changes or less, 2 changes or less, or 1 change compared to the CH2 domain sequence presented in SEQ ID NO: 5 or 6.

According to another preferred embodiment, the specific binding member comprises or consists of a sequence with one or more amino acid sequence changes (addition, deletion, substitution and/or insertion of an amino acid residue), preferably 40 changes or less, 30 changes or less, 20 changes or less, 15 changes or less, 10 changes or less, 5 changes or less, 4 changes or less, 3 changes or less, 2 changes or less, or 1 change compared to the Fcab sequence shown in SEQ ID NO: 23, 32, 50, 104, 77, 41, 59, 68, 86, 95, 15, 141, 132, 150, 123, 159, 167, 114, 25, 34, 52, 106, 79, 43, 61, 70, 88, 97, 16, 143, 134, 152, 125, 161, 169 or 116.

In case the specific binding member comprises a variant of the first sequence, the AB structural loop sequence, the CH3 domain, the Fcab or the heavy chain sequence disclosed in the present application, the specific binding member preferably retains the PPY sequence between positions 11 and 19, preferably positions 15 and 17, of the CH3 domain of the specific binding member. Furthermore, the specific binding member preferably retains an insert, preferably an insert of 5 amino acids between positions 16 and 17 of the CH3 domain of the specific binding member. According to another preferred embodiment, the specific binding member preferably retains the sequence at positions 97 and 98 of the CH3 domain of the specific binding member.

In particular, the specific binding member may be (or the antibody molecule may comprise) a variant of the specific binding member FS22-053, wherein said variant:

(i) comprises one or more amino acid sequence changes (addition, deletion, substitution and/or insertion of an amino acid residue), preferably 20 changes or less, 15 changes or less, 10 changes or less, 5 changes or less, 4 changes or less, 3 changes or less, 2 changes or less, or 1 change compared to the sequence of the FS22-053 specific binding element disclosed in the present application; or

(ii) has at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% sequence identity to the sequence of the FS22-053 specific binding member disclosed herein; and

wherein said specific binding member or antibody molecule comprises a PPY sequence between positions 15 and 17 and optionally comprises an insertion of 5 amino acids between positions 16 and 17 of the CH3 domain of the specific binding member or antibody molecule; and

Moreover, the indicated numbering of residues corresponds to the IMGT residue numbering scheme.

In addition, or alternatively, if the specific binding member comprises a variant of the CH3 domain, the CH2 and CH3 domain, Fcab, light chain or heavy chain sequence disclosed in the present application, the variant preferably does not comprise any amino acid changes in the first, second and third sequence located in the structural loops AB, CD and EF of the CH3 domain of the specific binding member. For example, the variant may not comprise any amino acid changes in the structural loops AB, CD and EF of the CH3 domain of the specific binding member.

In preferred embodiments, wherein one or more amino acids are replaced by another amino acid, the substitutions may be conservative substitutions, such as those in the following Table. In some embodiments, amino acids in the same category in the middle column are replaced with each other, i.e., for example, an apolar amino acid is replaced with another apolar amino acid. In some embodiments, amino acids in the same row in the rightmost column are replaced with each other.

Aliphatic Non-polar G A P I L V Polar - uncharged C S T M N Q Polar - charged D E K R Aromatic H F W Y

In some embodiments, the substitution(s) may be functionally conservative. In other words, in some embodiments, the substitution may not affect (or may not significantly affect) one or more functional properties (e.g., binding affinity) of a specific binding member containing the substitution, compared to an equivalent unsubstituted specific binding member.

Also provided is a specific binding member that comprises an antigen-binding site for CD137 located in a constant domain, preferably in the CH3 domain, of the specific binding member and that competes with the specific binding member of the present invention for binding to CD137 or that binds to the same epitope on CD137 as the specific binding member of the present invention. Methods for determining competition for an antigen between two specific binding members are known in the art. For example, competition for binding to an antigen between two specific binding members can be determined using surface plasmon resonance, such as Biacore. Methods for mapping an epitope bound by a specific binding member are also known in the art.

In some embodiments, the specific binding element may not comprise a CDR-based antigen-binding site.

In particular, the specific binding element may not contain a CDR-based antigen-binding site that binds PD-L1.

Additionally, or alternatively, the specific binding element may not contain a CDR-based antigen-binding site that binds mesothelin (MSLN).

For example, a specific binding member may not comprise a CDR-based antigen-binding site that binds PD-L1 or MSLN, wherein said specific binding member comprises a first, second and third sequence located within the AB, CD and EF structural loops of the CH3 domain of the specific binding member FS22-53-008 or FS22-172-003, the complete sequences of the AB and EF structural loops of the CH3 domain of the specific binding member FS22-53-008 or FS22-172-003, and/or the CH3 domain sequence of the specific binding member FS22-53-008 or FS22-172-003.

In particular, the specific binding element may not comprise the CDRs, VH and/or VL domains, and/or the heavy and/or light chain sequences of FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 shown below.

VH domain CDR FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

HCDR1(IMGT)GYPFTSYG

HCDR1 (Kabat) SYGIS

HCDR2(IMGT)ISAYSGGT

HCDR2 (Kabat) WISAYSGGTNYAQKLQG

HCDR3 (IMGT) ARDLFPTIFGVSYYYY

HCDR3 (Kabat) DLFPTIFGVSYYYY

VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

EVQLVQSGAEVKRPGASVKVSCKASGYPFTSYGISWVRQAPGQG

LEWMGWISAYSGGTNYAQKLQGRVTMTTDTSTSTAYMELRSLRS

DDTAVYYCARDLFPTIFGVSYYYYWGQGTLVTVSS

VL domain CDR FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

LCDR1 (IMGT) QSIGNR

LCDR1 (Kabat) RASQSIGNRLA

LCDR2 (IMGT) EAS

LCDR2 (Kabat) EASTSET

LCDR3 (IMGT) QQSYSTPYT

LCDR3 (Kabat) QQSYSTPYT

VL domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

DIQMTQSPSTLSASVRDRVIITCRASQSIGNRLAWYQHKPGKAPKL

LIYEASTSETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSY

STPYTFGQGTKLEIK

Heavy Chain FS22-172-003-AA/E12v2

EVQLVQSGAEVKRPGASVKVSCKASGYPFTSYGISWVRQAPGQGLEWMGWISAYSGGTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLFPTIFGVSYYYYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Light chain FS22-172-003-AA/E12v2

DIQMTQSPSTLSASVRDRVIITCRASQSIGNRLAWYQHKPGKAPKLLIYEASTSETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC

Heavy Chain FS22-053-008-AA/E12v2

EVQLVQSGAEVKRPGASVKVSCKASGYPFTSYGISWVRQAPGQGLEWMGWISAYSGGTNYAQKLQGRVTMTTDTSTSTAYMELRSLRSDDTAVYYCARDLFPTIFGVSYYYYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Light chain FS22-053-008-AA/E12v2

DIQMTQSPSTLSASVRDRVIITCRASQSIGNRLAWYQHKPGKAPKLLIYEASTSETGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC

Additionally or alternatively, the specific binding member may not comprise the CDRs and/or VH and/or VL domains of the anti-MSLN antibody, FS28-256-271, shown below.

CDR domain VH FS28-256-271

HCDR1(AA)(IMGT)GFTFTHTY

HCDR1 (AA) (Kabat) HTYMS

HCDR2 (AA) (IMGT) ISPTYSTT

HCDR2 (AA) Kabat) AISPTYSTTNYADSVKG

HCDR3 (AA) (IMGT) ARYNAYHAALDY

HCDR3 (AA) (Kabat) YNAYHAALDY

Domain VH FS28-256-271

EVQLLESGGGLVQPGGSLRLSCAASGFTFTHTYMSWVRQAPGKGLEWVSNISPTYSTTNYADSVKGRFTISRDNNKNTLYLQMNSLRAEDTAVYYCARYNAYHAALDYWGQGTLVTVSS

CDR domain VL FS28-256-271

LCDR1 (AA) (IMGT) QSVSSSY

LCDR1 (AA) (Kabat) RASQSVSSSYLA

LCDR2 (AA) (IMGT) GAS

LCDR2 (AA) (Kabat) GASSRAT

LCDR3 (AA) (IMGT) QQTVPYPYT

LCDR3 (AA) (Kabat) QQTVPYPYT

Domain VL FS28-256-271

EIVLTQSPGTLLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQTVPYPYTFGQGTKVEIK

The specific binding element may be conjugated to a bioactive molecule or a detectable label. In this case, the specific binding element may be called a conjugate. Such conjugates can be used in the treatment of the diseases described in this application.

For example, the bioactive molecule may be an immune system modulator, such as a cytokine, preferably a human cytokine. For example, the cytokine may be a cytokine that stimulates the activation and/or proliferation of T cells. Examples of cytokines for conjugation to a specific binding member include IL-2, IL-10, IL-12, IL-15, IL-21, GM-CSF, and IFN-gamma.

Alternatively, the bioactive molecule may be a ligand decoy, such as a cytokine ligand decoy, e.g., TGF-beta or IL-6.

Suitable detectable labels that can be conjugated to specific binding members are known in the art and include radioisotopes such as iodine-125, iodine-131, yttrium-90, indium-111 and technetium-99; fluorochromes such as fluorescein, rhodamine, phycoerythrin, Texas red and cyanine dye derivatives such as Cy7 and Alexa750; chromogenic dyes such as diaminobenzidine; latex beads; enzymatic labels such as horseradish peroxidase; phosphorus or laser dyes with spectrally isolated absorption or emission characteristics; and chemical groups such as biotin that can be detected by binding to a specific cognate detectable group, such as labeled avidin.

The specific binding member may be conjugated to the bioactive molecule or detectable label via any suitable covalent or non-covalent bond, such as a disulfide or peptide bond. If the bioactive molecule is a cytokine, the cytokine may be attached to the specific binding member via a peptide linker. Suitable peptide linkers are known in the art and may be 5 to 25, 5 to 20, 5 to 15, 10 to 25, 10 to 20, or 10 to 15 amino acids in length.

In some embodiments, the bioactive molecule may be conjugated to a specific binding member using a cleavable linker. The linker may provide for the release of the bioactive molecule from the specific binding member at the site of therapy. Linkers may include amide bonds (e.g., peptide linkers), disulfide bonds, or hydrazones. Peptide linkers, for example, may be cleaved by site-specific proteases, disulfide bonds may be cleaved by the reducing environment of the cytosol, and hydrazones may be cleaved by acid-mediated hydrolysis.

The conjugate may be a fusion protein comprising a specific binding element and a bioactive molecule. In this case, the bioactive molecule may be conjugated to the specific binding element via a peptide linker or a peptide bond. If the specific binding element is a multi-chain molecule, for example, if the specific binding element is or comprises Fcab or is MAB ² , the bioactive molecule may be conjugated to one or more chains of the specific binding element. For example, the bioactive molecule may be conjugated to one or both heavy chains of the MAB ² molecule. The advantage of fusion proteins is that they are easier to prepare and purify, facilitating the production of clinical grade materials.

The present invention also provides an isolated nucleic acid molecule or molecules encoding a specific binding member of the present invention. Such nucleic acid molecules are readily obtainable by a skilled artisan using methods well known in the art.

According to a preferred embodiment, the nucleic acid molecule encodes the CH3 domain of a specific binding element of: FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, more preferably FS22-172-003, FS22-172-002 or FS22-172-004, even more preferably FS22-172-003.

According to an alternative preferred embodiment, the nucleic acid molecule encodes the CH3 domain of a specific binding member of: FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably FS22-053-008.

The CH3 domain sequences of these specific binding elements are described in this application.

For example, a nucleic acid molecule that encodes the CH3 domain of a specific binding element:

(i) FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053 as depicted in SEQ ID NOs: 22, 31, 49, 103, 76, 40, 58, 67, 85, 94 and 176, respectively; or

(ii) FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172 as shown in SEQ ID NO: 140, 131, 149, 122, 158, 166 and 113, respectively.

According to a preferred embodiment, the nucleic acid molecule encodes a specific binding member of: FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, preferably FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005 or FS22-172-006, more preferably FS22-172-003, FS22-172-002 or FS22-172-004, even more preferably FS22-172-003.

According to an alternative preferred embodiment, the nucleic acid molecule encodes a specific binding member of: FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, preferably FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017 or FS22-053-014, more preferably FS22-053-008, FS22-053-009, FS22-053-011 or FS22-053-017, even more preferably FS22-053-008.

For example, a nucleic acid molecule that encodes a specific binding element:

(i) FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053 as depicted in SEQ ID NOs: 24, 33, 51, 105, 78, 42, 60, 69, 87, 96 and 177, respectively; or

(ii) FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172 as shown in SEQ ID NO: 142, 133, 151, 124, 160, 168 and 115, respectively.

The isolated nucleic acid molecule can be used to express a specific binding member according to the present invention. The nucleic acid will typically be provided in the form of a recombinant expression vector. Thus, according to another aspect of the present invention, there is provided a vector comprising a nucleic acid as described above. Suitable vectors can be selected or constructed containing appropriate regulatory sequences, including promoter sequences, terminator fragments, polyadenylation sequences, enhancer sequences, marker genes and other sequences, as appropriate. Preferably, the vector contains appropriate regulatory sequences for directing expression of the nucleic acid in a host cell. The vectors can be plasmids, viral vectors, for example, a phage, or a phagemid, as appropriate.

The nucleic acid molecule or vector described in this application can be introduced into a host cell. Techniques for introducing nucleic acid or vectors into host cells are well known in the art, and any suitable technique can be used. A number of host cells suitable for producing recombinant specific binding members are known in the art and include bacterial, yeast, insect or mammalian host cells. A preferred host cell is a mammalian cell, such as a CHO, NS0 or HEK cell, for example a HEK293 cell. Most preferred host cell is a CHO cell.

According to another aspect of the present invention, there is provided a method for producing a specific binding member according to the present invention, comprising expressing a nucleic acid encoding a specific binding member in a host cell and optionally isolating and/or purifying the specific binding member thus obtained. Methods for culturing host cells are well known in the art. The method may further comprise isolating and/or purifying the specific binding member. Techniques for purifying recombinant specific binding members are well known in the art and include, for example, HPLC, FPLC or affinity chromatography, for example using protein A or protein L. In some embodiments, purification can be accomplished using an affinity tag on the specific binding member. The method may also comprise preparing the specific binding member as a pharmaceutical composition, optionally with a pharmaceutically acceptable excipient or other substance described below.

As explained above, CD137 is expressed on cells of the immune system, including CD8 ⁺ T cells, CD4 ⁺ T cells, Treg cells, B cells, NK cells, NKT cells, dendritic cells, and tumor-infiltrating lymphocytes (TILs). Specifically, CD137 upregulation has been shown to play a role in enhancing CD8 ⁺ T cell proliferation, survival, and cytotoxic effector function, as well as in CD8 ⁺ T cell differentiation and the maintenance of memory CD8 ⁺ T cells. CD137 is expressed at lower levels on CD4 ⁺ T cells than on CD8 ⁺ T cells, but has also been shown to be involved in the induction of proliferation and activation of some CD4 ⁺ T cell subsets. CD137 activation has also been shown to enhance NK cell-mediated ADCC, as well as B cell proliferation, survival, and cytokine production.

Given the activity of CD137 in enhancing the immune response, CD137 agonist molecules have been studied for the treatment of cancer as well as the treatment of chronic infections.

Thus, the specific binding elements described in the present application can be used for therapeutic applications, in particular in the treatment of cancer. In addition, it is expected that the specific binding elements can be used in the treatment of infectious diseases, such as persistent infectious diseases.

The specific binding element described in the present application can be used in a method of treating a human or animal organism. According to related aspects of the present invention, there is provided;

(i) a specific binding element described in this application for use as a medicinal product,

(ii) a specific binding element described in this application for use in a method of treating a disease or disorder,

(iii) the use of a specific binding member described herein in the manufacture of a medicament for use in the treatment of a disease or disorder; and,

(iv) a method of treating a disease or disorder in an individual, said method comprising administering to the individual a therapeutically effective amount of a specific binding member described herein.

The individual may be a patient, preferably a human patient.

Treatment may be any treatment or therapy that achieves some intended therapeutic effect, such as inhibition or delay of progression of the condition, and includes decreasing the rate of progression, slowing the rate of progression, ameliorating the condition, curing or causing remission (whether partial or complete) of the condition, preventing, ameliorating, delaying, alleviating or reversing one or more symptoms and/or signs of the condition, or prolonging the survival of an individual or patient beyond that expected in the absence of treatment.

Also included is treatment as a prophylactic measure (e.g., prevention). For example, an individual susceptible to or at risk of developing or recurring a disease, such as cancer, may be treated as described herein. Such treatment may prevent or delay the development or recurrence of the disease in the individual.

The described method of treatment may include administering at least one additional treatment agent to the individual in addition to the specific binding member. Thus, the specific binding member described herein may be administered to the individual alone or in combination with one or more other treatment agents. If the specific binding member is administered to the individual in combination with another treatment, the additional treatment may be administered to the individual simultaneously, sequentially, or separately from the administration of the specific binding member. If the additional treatment is administered simultaneously with the specific binding member, the specific binding member and the additional treatment may be administered to the individual as a combination preparation. For example, the additional therapy may be a known therapy or therapeutic agent for the disease being treated.

Although the specific binding element may be administered alone, the specific binding elements will typically be administered in the form of a pharmaceutical composition, which may contain at least one component in addition to the specific binding element. Thus, according to another aspect of the present invention, there is provided a pharmaceutical composition comprising the specific binding element described herein. Also provided is a method comprising preparing the specific binding element in the form of a pharmaceutical composition.

The pharmaceutical compositions may contain, in addition to the specific binding element, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer, or other materials well known to those skilled in the art. As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions, and/or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of a subject (e.g., a human) without undue toxicity, irritation, allergic reaction, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. The precise nature of the carrier or other material will depend on the route of administration, which may be by infusion, injection, or any other suitable route, as discussed below.

For parenteral, e.g. subcutaneous or intravenous administration, e.g. by injection, the pharmaceutical composition containing the specific binding element may be in the form of a parenterally acceptable aqueous solution which is free of pyrogens and has a suitable pH, isotonicity and stability. Those skilled in the art will be well able to prepare suitable solutions using, for example, isotonic vehicles such as sodium chloride injection, Ringer's injection, lactated Ringer's injection. If necessary, preservatives, stabilizers, buffers, antioxidants and/or other additives may be used, including buffers such as phosphate, citrate and other organic acids; antioxidants such as ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3'-pentanol and m-cresol); low molecular weight polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagines, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or nonionic surfactants such as Tween™, Pluronic Agents™, or polyethyleneglycol (PEG).

In some embodiments, the specific binding elements may be provided in lyophilized form for reconstitution prior to administration. For example, the lyophilized specific binding elements may be reconstituted in sterile water and mixed with physiological saline prior to administration to an individual.

Administration may be in a "therapeutically effective amount" which is sufficient to show benefit to the individual. The actual amount administered and the rate and duration of administration will depend on the nature and severity of what is being treated, the particular individual being treated, the clinical condition of the individual, the cause of the disorder, the site of delivery of the composition, the type of specific binding member, the route of administration, the schedule of administration and other factors known to practitioners. The administration of treatment, such as dosage decisions, etc., is the responsibility of general practitioners and other physicians and may depend on the severity of the symptoms and/or progression of the disease being treated. Appropriate doses of immunoglobulins are well known in the art (Ledermann et al. (1991) Int. J. Cancer 47: 659-664; and Bagshawe et al. (1991) Antibody, Immunoconjugates and Radiopharmaceuticals 4: 915-922). Specific dosages as described herein or in the Physician's Desk Reference (2003) appropriate to the antibody molecule being administered may be used. In the case of antibody molecules, a therapeutically effective amount or appropriate dose of the specific binding member may be determined by comparing in vitro activity with in vivo activity in an animal model. Methods are known in the art for extrapolating effective dosages in mice and other test animals to humans. The precise dosage will depend on a number of factors, including the size and location of the area to be treated and the particular nature of the specific binding member.

A typical immunoglobulin dose ranges from 100 mcg to 1 g for various systemic applications and from 1 mcg to 1 mg for various topical applications. A higher initial loading dose may be given followed by one or more lower doses. This is the single adult dose, which may be proportionally adjusted for children and infants, and adjusted for other specific binding element formats proportionally to molecular weight.

Treatments may be repeated at daily, twice weekly, weekly or monthly intervals at the discretion of the physician. The treatment schedule for an individual may depend on the pharmacokinetic and pharmacodynamic properties of the specific binding member composition, the route of administration and the nature of the condition being treated.

The treatment may be intermittent, and the period between administrations may be about two weeks or more, such as about three weeks or more, about four weeks or more, about once a month or more, about five weeks or more, or about six weeks or more. For example, the treatment may be administered every two to four weeks or every four to eight weeks. Suitable formulations and routes of administration are described above.

According to a preferred embodiment, the specific binding element described in the present application can be used in a method of treating cancer.

Cancer may be characterized by abnormal proliferation of malignant cancer cells. When a specific type of cancer, such as breast cancer, is mentioned, this refers to abnormal proliferation of malignant cells of the corresponding tissue, such as breast tissue. A secondary cancer that is localized in the breast but is the result of abnormal proliferation of malignant cells of another tissue, such as ovarian tissue, is not breast cancer as referred to in this application, but ovarian cancer.

The cancer may be a primary or secondary cancer. Thus, the specific binding element described in the present application can be used in a method of treating cancer in an individual, wherein said cancer is a primary tumor and/or a tumor metastasis.

A cancer tumor to be treated using a specific binding member described in the present application may contain TILs that express CD137, for example, on its cell surface. According to one embodiment, the tumor could be determined to contain TILs that express CD137. Methods for determining antigen expression on a cell surface are known in the art and include, for example, flow cytometry.

For example, a cancer to be treated using a specific binding member described herein may be selected from the group consisting of leukemias such as acute myelogenous leukemia (AML), chronic myelogenous leukemia (CML), acute lymphoblastic leukemia (ALL), and chronic lymphocytic leukemia (CLL); lymphomas such as Hodgkin's lymphoma, non-Hodgkin's lymphoma, and multiple myeloma; and different types of solid cancer such as sarcomas (eg, soft tissue sarcomas), skin cancer (eg, Merkel cell carcinoma), melanoma, bladder cancer (eg, urothelial carcinoma), brain cancer (eg, glioblastoma multiforme), breast cancer, uterine/endometrial cancer, ovarian cancer (eg, ovarian serous cystadenoma), prostate cancer, lung cancer (eg, non-small cell lung carcinoma (NSCLC) and small cell lung cancer (SCLC)), colorectal cancer (eg, colorectal adenocarcinoma), cervical cancer (eg, squamous cell carcinoma of the cervix and adenocarcinoma of the cervix), liver cancer (eg, hepatocellular carcinoma), head and neck cancer (eg, squamous cell carcinoma of the head and neck), esophageal cancer, pancreatic cancer, kidney cancer (eg, renal cell carcinoma), adrenal gland cancer, stomach cancer, testicular cancer, gallbladder and bile duct cancer (eg, cholangiocarcinoma), thyroid cancer, thymus cancer, bone cancer, and brain cancer.

According to a preferred embodiment, the cancer to be treated using the specific binding member described in the present application is a solid cancer. More preferably, the cancer to be treated using the specific binding member described in the present application is a solid cancer selected from the group consisting of: sarcoma, melanoma, bladder cancer, brain cancer, breast cancer, uterine/endometrial cancer, ovarian cancer, prostate cancer, lung cancer, colorectal cancer, cervical cancer, liver cancer, head and neck cancer, pancreatic cancer, renal cancer, and gastric cancer.

In the context of cancer, treatment may include inhibition of cancer growth, including complete remission of cancer, and/or inhibition of cancer metastasis, and inhibition of cancer recurrence. Cancer growth generally refers to any of a number of indicators that indicate a change within the cancer to a more advanced form. Thus, indicators for measuring inhibition of cancer growth include a decrease in cancer cell survival, a decrease in tumor volume or morphology (e.g., as determined using computed tomography (CT), sonography, or other imaging modality), delayed tumor growth, destruction of tumor vasculature, improved delayed-type hypersensitivity skin test results, increased activity of anti-cancer immune cells or other anti-cancer immune responses, and a decrease in levels of tumor-specific antigens. Activation or enhancement of immune responses to cancers in an individual may improve the individual's ability to resist cancer growth, particularly the growth of cancer already present in the subject, and/or reduce the propensity for cancer growth in the individual.

In the context of cancer treatment, a specific binding member described herein may be administered to an individual in combination with another cancer therapy or therapeutic agent, such as an cancer therapy or therapeutic agent that has been shown to be, or is expected to be, suitable for treating the cancer in question. For example, a specific binding member may be administered to an individual in combination with a chemotherapeutic agent, radiation therapy, an immunotherapeutic agent, a tumor vaccine, an oncolytic virus, an adoptive cell transfer (ACT) therapy (such as adoptive NK cell therapy or chimeric antigen receptor (CAR) T cell therapy, autologous tumor infiltrating lymphocytes (TILs), or gamma/delta T cells), or a hormonal therapy agent.

Without being bound by any theory, it is believed that the specific binding element described in the present application can act as an adjuvant in anti-cancer therapy. In particular, it is believed that the administration of the specific binding element to an individual in combination with chemotherapy and/or radiation therapy or in combination with an anti-tumor vaccine, for example, will induce a stronger immune response against cancer than that achieved by using chemotherapy and/or radiation therapy or an anti-tumor vaccine alone.

One or more chemotherapeutic agents to be administered in combination with the specific binding member described herein may be selected from the group consisting of: taxanes, cytotoxic antibiotics, tyrosine kinase inhibitors, PARP inhibitors, B-Raf enzyme inhibitors, MEK inhibitors, c-MET inhibitors, VEGFR inhibitors, PDGFR inhibitors, alkylating agents, platinum analogs, nucleoside analogs, antifolates, thalidomide derivatives, antineoplastic chemotherapeutic agents, and others. Taxanes include docetaxel, paclitaxel, and nab-paclitaxel; cytotoxic antibiotics include actinomycin, bleomycin, and anthracyclines such as doxorubicin, mitoxantrone, and valrubicin; Tyrosine kinase inhibitors include erlotinib, gefitinib, axitinib, PLX3397, imatinib, cobemitinib and trametinib; PARP inhibitors include piraparib; B-Raf enzyme inhibitors include vemurafenib and dabrafenib; alkylating agents include dacarbazine, cyclophosphamide and temozolomide; platinum analogues include carboplatin, cisplatin and oxaliplatin; nucleoside analogues include azacitidine, capecitabine, fludarabine, fluorouracil and gemcitabine; antifolates include methotrexate and pemetrexed. Other chemotherapeutic agents suitable for use in the present invention include defactinib, entinostat, eribulin, irinotecan and vinblastine.

Preferred therapeutic agents for administration with the antibody molecule described herein are doxorubicin, mitoxantrone, cyclophosphamide, cisplatin and oxaliplatin.

The radiation therapy to be administered in combination with the specific binding element described in this application may be external beam radiation therapy or brachytherapy.

An immunotherapeutic agent for administration in combination with a specific binding member described herein may be a therapeutic antibody molecule, a nucleic acid, a cytokine, or a cytokine-based therapy. For example, a therapeutic antibody molecule may bind to an immune regulatory molecule, such as a checkpoint inhibitor molecule or an immune costimulatory molecule, or to a tumor antigen, such as a cell surface tumor antigen or a soluble tumor antigen. Examples of immune regulatory molecules that a therapeutic antibody molecule may bind to include CTLA-4, LAG-3, TIGIT, TIM-3, VISTA, PD-L1, PD-1, CD47, CD73, CSF-1R, KIR, CD40, HVEM, IL-10, and CSF-1. Examples of innate immune receptors that a therapeutic antibody molecule may bind to include TLR1, TLR2, TLR4, TLR5, TLR7, TLR9, RIG-I-like receptors (e.g., RIG-I and MDA-5), and STING. Examples of tumor antigens that a therapeutic antibody molecule may bind to include HER2, EGFR, CD20, and TGF-beta.

The nucleic acid to be administered in combination with the specific binding element described herein may be siRNA.

The cytokine or cytokine-based therapy may be selected from the group consisting of: IL-2, a prodrug of conjugated IL-2, GM-CSF, IL-7, IL-12, IL-9, IL-15, IL-18, IL-21, and interferon type I.

Tumor vaccines for cancer treatment have been introduced into the clinic and are discussed in detail in the scientific literature (e.g. Rosenberg, S. 2000). They mainly focus on strategies to stimulate the immune system response to various cellular markers expressed by autologous or allogeneic cancer cells by using these cells as a vaccination route, either with or without granulocyte-macrophage colony-stimulating factor (GM-CSF). GM-CSF induces a strong response upon antigen presentation and works particularly well when used with these strategies.

The chemotherapeutic agent, radiation therapy, immunotherapeutic agent, tumor vaccine, oncolytic virus, ACT therapy or hormonal therapy agent is preferably a chemotherapeutic agent, radiation therapy, immunotherapeutic agent, tumor vaccine, oncolytic virus, ACT therapy or hormonal therapy agent for the cancer in question, i.e., a chemotherapeutic agent, radiation therapy, immunotherapeutic agent, tumor vaccine, oncolytic virus, ACT therapy or hormonal therapy agent that has been shown to be effective in the treatment of the cancer in question. The selection of an appropriate chemotherapeutic agent, radiation therapy, immunotherapeutic agent, tumor vaccine, oncolytic virus, ACT therapy or hormonal therapy agent that has been shown to be effective for the cancer in question is within the capabilities of a skilled practitioner.

Given the activity of CD137 in enhancing the immune response, CD137 agonist molecules are expected to be useful in the treatment of infectious diseases. Thus, in another preferred embodiment, the antibody molecule described herein can be used in a method for treating an infectious disease, such as an acute or persistent infectious disease.

Without being limited by any theory, it is believed that CD137 agonist molecules may be able to enhance the immune response against an acute infectious disease caused by a pathogen by inducing rapid infiltration and activation of innate immune cells such as neutrophils and monocytes, thereby facilitating the clearance of the pathogen that caused the acute infectious disease. Thus, according to a further embodiment, the antibody molecule described in the present application can be used in a method for treating an acute infectious disease, such as an acute bacterial disease. According to a preferred embodiment, the acute infectious disease is an acute bacterial disease caused by infection with a gram-positive bacterium, such as a bacterium of the genus Listeria, Streptococcus pneumoniae or Staphylococcus aureus.

Infectious diseases are usually cleared by the immune system, but some infections persist for long periods of time, such as months or years, and the immune system is ineffective at fighting them. Such infections are also called persistent or chronic infections.

Preferably, the antibody molecule described herein is used to treat a persistent infectious disease, such as a persistent viral, bacterial, fungal or parasitic infection, preferably a persistent viral or bacterial infection.

According to a preferred embodiment, the persistent viral infection to be treated using the antibody molecule described herein is a persistent infection caused by: human immunodeficiency virus (HIV), Epstein-Barr virus, cytomegalovirus, hepatitis B virus, hepatitis C virus, or varicella zoster virus.

According to a preferred embodiment, the persistent bacterial infection to be treated using the antibody molecule described herein is a persistent infection of: Staphylococcus aureus, Hemophilus influenza, Mycobacterium tuberculosis, Mycobacterium leprae, Salmonella typhi, Helicobacter pylori, Treponema pallidum or Streptococcus pneumoniae.

It has been described that CD137 agonism is beneficial in the treatment of gram-positive bacterial infections. Thus, in a preferred embodiment, the persistent bacterial infection to be treated using the antibody molecule described herein is a persistent infection with a gram-positive bacterium. In a more preferred embodiment, the persistent bacterial infection is a persistent infection with a gram-positive bacterium selected from the group consisting of Staphylococcus aureus, Mycobacterium leprae and Streptococcus pneumoniae.

According to a preferred embodiment, the persistent fungal infection to be treated using the specific binding member described herein is a persistent infection of Candida (e.g. Candida albicans), Cryptococcus (e.g. Cryptococcus gattii or Cryptococcus neoformans), Talaromyces (Penicillium) (e.g. Talaromyces marneffe), Microsporum (e.g. Microsporum audouinii) or Trichophyton tonsurans.

According to a preferred embodiment, the persistent parasitic infection to be treated using the specific binding member described herein is a persistent infection with Plasmodium, such as Plasmodium falciparum, or Leishmania, such as Leishmania donovani.

When applied to the treatment of a persistent infectious disease, treatment may include eliminating the infection, reducing the individual's pathogen load, or preventing recurrence of the infection. For example, treatment may include preventing, ameliorating, delaying, reducing, or stopping one or more symptoms and/or signs of a persistent infection. Alternatively, treatment may include preventing the infectious disease.

In the context of treating infectious diseases, the specific binding member described herein can be administered to an individual in combination with another therapeutic agent for treating an infectious disease, such as a therapeutic agent that has been shown to be suitable, or is expected to be suitable, for treating the infectious disease in question. For example, the specific binding member can be administered to an individual in combination with an immunotherapeutic agent. The immunotherapeutic agent for administration in combination with the antibody molecule described herein can be a therapeutic antibody molecule. For example, the therapeutic antibody molecule can bind to a receptor of the innate immune system. Examples of receptors of the innate immune system that a therapeutic antibody molecule can bind to include TLR1, TLR2, TLR4, TLR5, TLR7, TLR9, RIG-I-like receptors (e.g., RIG-I and MDA-5), and STING.

If the specific binding element is used to prevent an infectious disease, the specific binding element may be administered in combination with a vaccine against the pathogen in question. Without wishing to be bound by any theory, it is believed that the specific binding element described herein may act as an adjuvant in vaccination. In particular, it is believed that administration of the specific binding element to an individual in combination with a vaccine will elicit a stronger immune response against the pathogen than that achieved by using the vaccine alone.

Other aspects and embodiments of the present invention will be apparent to those skilled in the art based on the present disclosure, including the following experimental examples.

All documents referenced in this specification are incorporated into this application by reference in their entirety.

In this application, "and/or" should be understood as a specific disclosure of each of the two stated features or components, with or without the other. For example, "A and/or B" should be understood as a specific disclosure of each of (i) A, (ii) B, and (iii) A and B, as if each of them were presented separately in this application.

Unless the context dictates otherwise, the descriptions and definitions of features provided above are not limited to any particular aspect or embodiment of the present invention and are equally applicable to all aspects and embodiments described.

Other aspects and embodiments of the present invention provide the aspects and embodiments described above with the term "comprising" replaced by the term "consisting of" or "consisting essentially of" unless the context dictates otherwise.

Certain aspects and embodiments of the present invention will now be illustrated by way of example and with reference to the figures described above.

Examples

Example 1: Production, characterization and selection of human, mouse and cynomolgus monkey antigens

1.1 Recombinant antigens

Members of the tumor necrosis factor receptor superfamily (TNFRSF) are known for their propensity to form multimers that cluster when bound to their cognate ligands (Croft, M. 2003). This propensity to aggregate to achieve functionality makes it difficult to produce soluble recombinant proteins that do not aggregate in solution for use in in vitro selections, such as phage and yeast display, and for characterization of selected proteins.

Several commercially available recombinant antigens were tested and most were found to be unsuitable for use in such selections due to the levels of aggregates present. Of those tested, only biotinylated human CD137 secreted protein fused to hFc (BPS Biosciences, Cat#71171), hereafter designated "hCD137-hFc-Avi-BPS", had low enough aggregation to be suitable and was used in selections, but with limited success (see Example 2).

Since most commercially available antigens were deemed unsuitable, the following recombinant dimeric and monomeric CD137 antigens (see Table 1) were produced in-house for use in screening:

Table 1

Type Designation View Soluble or cellular Biotinylated Antigen format Recombinant mCD137-mFc-Avi Mouse Soluble Yes Dimer Recombinant mCD137-Avi-His Mouse Soluble Yes Monomer Recombinant hCD137-mFc-Avi Human Soluble Yes Dimer Recombinant hCD137-Avi-His Human Soluble Yes Monomer Recombinant cCD137-mFc-Avi Javan macaque Soluble Yes Dimer

Monomeric antigens were generated by cloning DNA encoding the extracellular domain of human (SEQ ID NO: 181) or mouse (SEQ ID NO: 185) CD137, together with the Avi sequence and six C-terminal histidine residues, into modified pFUSE vectors (Invivogen, catalog # pfuse-mg2afc2) using EcoRI-HF and BamHI-HF restriction enzymes. Vectors were transfected into HEK293-6E cells (National Research Council Canada) and expressed CD137 was purified using a HisTrap™ excel nickel column (GE LifeSciences 29048586) and size exclusion chromatography (SEC) to ensure that the antigen was a single molecule and did not contain aggregates.

To generate dimeric antigens, DNA constructs encoding the extracellular domain of human, mouse, or cynomolgus monkey CD137 fused to the Fc domain of mIgG2a along with the Avi sequence were cloned into modified pFUSE vectors and transfected into HEK293-6E cells. Recombinant CD137 was purified using a MabSelect SuRe™ Protein A column (GE Healthcare, 11003494) and size exclusion chromatography (SEC) to ensure that the antigen was a single molecule and did not contain aggregates.

Each of the dimeric and monomeric antigens was biotinylated using the BirA Biotin-Protein Ligase Reaction Kit (Avidity LLC, BirA500) to generate monomeric CD137 antigens labeled with one biotin molecule and dimeric CD137 antigens labeled with two biotin molecules, one on each of the two monomers. 3 mg of antigen was mixed with 7.8 μl of the BirA enzyme mixture to a molar ratio of enzyme to substrate of 1:50. Supplements were then added according to the manufacturer's recommendations (142 μl Biomix A, 142 μl Biomix B, 142 μl biotin) and the reaction mixture was incubated for two hours at room temperature. The buffer in the reaction mixture was immediately replaced with DPBS (Life Technologies 14190-169) using 30 μm Amicon filters (Merck Millipore UFC503096).

Proteins were further purified by SEC to ensure removal of the BirA enzyme and to obtain a final high-quality monodisperse protein preparation free of high molecular weight aggregates. In detail, materials from the same production lot were mixed and analyzed for stability and purity by size exclusion high-performance liquid chromatography (SE-HPLC), SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography with multi-angle light scattering (SEC-MALS). Complete biotinylation of proteins was confirmed by streptavidin mobility shift SDS-PAGE. Binding of recombinant human and mouse antigens to positive control anti-CD137 antibodies (20H4.9 (US Patent No. 7,288,638) and Lob12.3 (University of Southampton), respectively) was confirmed in vitro by surface plasmon resonance (SPR), and binding to DO11.10 cells expressing human and mouse CD137 ligand was confirmed by flow cytometry. Cells were incubated with CD137 antigens for 1 h, and then a fluorescently labeled anti-mouse Fc antibody was used to detect cell binding. Binding of recombinant cynomolgus monkey antigen to DO11.10 cells (National Jewish Health) expressing cynomolgus monkey CD137 ligand was confirmed by flow cytometry as described above. To ensure the highest possible purity of materials used in the selection protocols, careful characterization of antigen proteins was performed to ensure that the presence of protein aggregates did not exceed 2%.

1.2 Antigens expressed by cells

DO11.10 cells (National Jewish Health) expressing full-length murine CD137 (SEQ ID NO: 184) or human CD137 (SEQ ID NO: 180), designated “DO11.10.mCD137” and “DO11.10.hCD137,” respectively, were generated to present the antigen in a membrane-bound conformation most similar to its native form for selection and further characterization of the selected Fcabs listed in Table 2.

Lentiviral transduction was used to generate DO11.10 cells overexpressing human or murine CD137 receptors using the Lenti-X HTX Packaging System (Takara, Cat. #631249). The Lenti-X expression vector (pLVX) (Takara, Cat. #631253) containing cDNA encoding human CD137 (SEQ ID NO: 180) or murine CD137 (SEQ ID NO: 184) was co-transfected using the Lenti-X HTX Packaging Mix into the Lenti-X 293T cell line (Takara, Cat. #632180) to produce virus. The DO11.10 cell line was then transduced with these lentiviral vectors.

Expression of human CD137 or mouse CD137 on these cells was confirmed by flow cytometry based on cell binding of CD137 antibodies, 20H4.9 and Lob12.3, used as positive controls, respectively. Cells were incubated with human or mouse positive control antibodies for 1 hour, and then fluorescently labeled anti-human Fc detection antibody (Stratech Scientific Ltd, cat. no. 109-546-098-JIR) was used to detect cell binding.

DO11.10 cells expressing cynomolgus monkey CD137, designated “DO11.10.cCD137”, were also generated using the same lentiviral transduction methodology and used to test the cross-reactivity of Fcab to human CD137 with cynomolgus monkey CD137. Expression of cynomolgus monkey CD137 was confirmed by flow cytometry based on binding to the cells of anti-CD137 antibody (MOR_7480.1, US 2012/0237498 A1) used as a positive control as described previously.

Table 2

Type Designation View Presentation Cell DO11.10.hCD137 Human Expressed by the cell Cell DO11.10.mCD137 Mouse Expressed by the cell Cell DO11.10.cCD137 Javan macaque Expressed by the cell

Example 2: Naive selection of Fcab to human CD137

To discover Fcabs that bind to human CD137 and to maximize the diversity of binders identified, both yeast display and phage display selection campaigns were employed. Because CD137 is expressed at low levels in a number of non-immune cell types, it was decided to select Fcabs to human CD137 that selectively target cells that express CD137 in high abundance, such as activated T cells. Without being bound by theory, it was hypothesized that cells with very low or negligible levels of CD137 expression would be more likely to have CD137 in a monomeric state on their cell surface, in contrast to activated T cells with highly upregulated CD137 expression, in which the majority of the protein would be expected to be present as dimers, trimers, or higher-order multimers on the cell surface.

Cells overexpressing CD137 or recombinant human dimeric CD137 proteins were used in the Fcab selections to target these representative epitopes, which facilitate binding interactions with cells with strong upregulation of multimeric CD137. In addition, the use of dimeric antigens was considered advantageous for the selection of bivalent Fcabs that avidly bound CD137 and remained bound to the target longer, which in turn was hypothesized to facilitate preferential binding to cells with upregulated CD137 expression levels. In this case, monomeric recombinant CD137 was not used to prevent the selection of very high affinity monovalent Fcab binders that might bind CD137 too strongly.

The goal of this selection strategy was to obtain Fcabs that would preferentially bind activated T cells and would not bind well to naive T cells expressing only monomeric CD137 or to other cells that express very low levels of CD137. By selecting CD137 Fcabs that bind bivalently and preferentially to dimeric and multimeric antigen over monomeric antigen, it was believed that potential off-target T cell activation would be reduced, with associated reduced toxicity.

Phage display

Six naive phage libraries displaying the CH3 domain of human IgG1 were used for phage display selections. All six libraries contained randomized AB loops (containing residues at positions 14-18 according to IMGT numbering) and randomized EF loops (containing residues at positions 92-101 according to IMGT numbering). One of the libraries contained clones with an insertion of either two or four amino acids (encoded by two or four NNK codons) at position 101 in the EF loop (the inserted residues are at positions 101.4-101.1 according to IMGT numbering).

A total of 12 selection campaigns were performed to identify binders to human CD137. In-house produced hCD137-mFc-Avi antigen or hCD137-hFc-Avi-BPS antigen from a commercial source and/or DO11.10.hCD137 cells expressing the antigen were used in selections using six phage libraries. Since only one functional Fcab sequence was identified in the initial selections using recombinant hCD137-hFc-Avi-BPS antigen, in-house produced hCD137-mFc-Avi was used in the remaining rounds of selections on recombinant CD137. Initial rounds were performed using 100 nM biotinylated antigen with a deselection step using 500 nM unlabeled recombinant human Fc fragment produced in-house. Phage binding elements were captured using magnetic beads coated with either streptavidin or neutravidin. For selection effluents in which non-specific binding to the Fc fragment was detected by phage ELISA (A450-630 nm greater than 0.4), an additional deselection step was introduced in which a biotinylated Fc fragment (produced in-house) was incubated with the phage effluents. Non-Fc binding elements were separated from Fc binding elements by magnetic capture of biotinylated Fc binding phage.

To detect binders to the membrane-bound native conformation of CD137, cell selections were performed as follows: phage effluents from recombinant antigen selections were incubated with human CD137-deficient DO11.10 cells to remove unwanted binders, such as those that bind non-specifically to the cells. Phage were then incubated with 1× ¹⁰⁷ DO11.10.hCD137 cells, binders were eluted by trypsin digestion, and then expanded for the next round. Round 3 was performed in a similar manner, increasing the selection pressure by decreasing the number of DO11.10.hCD137 cells to 5× ¹⁰⁶ .

Phage ELISA was performed after each round of selection to determine the enrichment of antigen-specific phage. 96-well streptavidin plates were coated overnight with 1 μg/ml biotinylated antigen. After blocking the plates with 4% marvel phosphate-buffered saline (PBS), 50 μl of phage-containing bacterial supernatant were added to each well and incubated for 1 h at room temperature with shaking at 450 rpm. The phage solution was removed by inverting the plate and washing 4 times with PBS containing 0.1% Tween and 4 times with PBS. HRP-conjugated anti-phage M13 antibody was then added to detect phage bound to the immobilized antigen. The phage solution was removed by inverting the plate and washing 4 times with PBS containing 0.1% Tween and 4 times with PBS. TMB peroxidase detection solution in microwells was added to each well and allowed to develop color for up to 30 minutes. The reaction was stopped with 1 M sulfuric acid and the plate was read at OD _450-630 in a microtiter plate reader. Specific candidates (“hits”) were defined as those with a signal intensity for recombinant CD137 that was at least 4-fold greater than background as determined by the negative control, i.e., PBS or a negative non-binding phage such as wild-type CH3, and at least 10-fold greater than the binding strength for biotinylated recombinant Fc fragment.

Phage fluorescence-activated cell sorting (FACS) analyses were performed on the cell selection effluents from rounds 2 and 3. In general, 2 x ¹⁰⁵ cells were transferred to a round-bottomed microtiter plate. Phage supernatant was added to the cells and incubated for 1 h at 4°C. Cells were then washed twice with ice-cold PBS containing 2% BSA, resuspended in 100 μl of anti-M13 and goat anti-mouse IgG F(ab') fragment conjugated to FITC, and incubated with the cells for 1 h on ice. Cells were analyzed in a flow cytometer after 3 washes in PBS. In FACS, phage-specific candidates were defined as those whose geometric mean fluorescence intensity (MFI) for CD137-positive cells was at least 10-fold higher than the binding signal to recombinant CD137 and at least 4-fold higher than the background determined by the negative control (PBS).

3230 phage clones were screened by phage ELISA for binding to dimeric recombinant hCD137 antigen, and recombinant Fc was used as a negative control. 1140 phage clones were screened by phage FACS for specific binding to DO11.10.hCD137 cells. Individual candidates were then sequenced and the resulting 76 unique sequences were assigned the clone identifier Fcab and subcloned into the pTT5 expression vector (National Research Council Canada) containing the HelD1.3 IgG1 heavy chain expression cassette to express the Fcab clones in the MAb ² format (see Example 3.1).

Yeast display

Four naive yeast libraries displaying the CH1 to CH3 domains of human IgG1 were used for yeast display selection. All four libraries contained randomized AB loops (containing residues at positions 14 to 18 according to IMGT numbering) and randomized EF loops (containing residues at positions 92 to 101 according to IMGT numbering) in the CH3 domain. Two of the libraries additionally contained a five-residue insertion at position 16 in the AB loop of the CH3 domain (residues at positions 16.5 to 16.1 according to IMGT numbering).

In-house produced hCD137-mFc-Avi antigen or hCD137-hFc-Avi-BPS antigen from a commercial source were used in selections using four yeast libraries, i.e., similar to the phage selections, antigen from the commercial source was not used after the initial rounds of selection and in-house produced antigen was used instead. For each library, a first round of selection was performed using magnetic cell separation (MACS). Libraries were grown, induced and 1 x 10 ¹⁰ cells were incubated with 250 or 300 nM biotinylated recombinant antigen after a deselection step using 1.25 μM unlabeled human or mouse Fc. Yeast binders were separated by the addition of streptavidin magnetic beads and using MACS LS columns (Miltenyi Biotech 130-042-401). Subsequent rounds of selections were performed by fluorescence-activated cell sorting (FACS) on FACS-Aria II instruments (BD Bioscience) using fluorescently labeled antibodies to detect bound antigen (anti-biotin-APC (Miltenyi Biotech 130-090-856), anti-streptavidin-APC (BD Biosciences 349024), or anti-neutravidin-DyLight-488 (Thermo Fisher 22832)), correctly folded IgG scaffolds (FITC-labeled anti-human IgG CH2 domain antibody (Biorad AbD Serotec (MCA647F)) or aga2-IgG expression construct (anti-Xpress (Life Technologies R91025) and FITC-labeled anti-mouse IgG antibody (Sigma F2653-.5ML)). Whenever possible, the antigen marker was used in combination with one of the structural markers to normalize the binding signal intensity to yeast surface Fcab expression. Unstained and control yeast populations were used to set up sorting gates as follows: yeast cells on the FSC-A and SSC-A plot, FSC-W and FSC-H plots to distinguish singlets from multiplets or budding yeast cells, and FITC-A and APC-A plots to detect Fcab-binding double-positive cells.

The antigen concentration used in each round was determined empirically by performing a quality control of the output using the antigen concentration used in the previous round. If binding enrichment increased compared to the previous round (>5-fold), the antigen concentration was reduced by 1:2 or 1:3, otherwise it was held constant. This method was also followed to determine the number of selection rounds required, as well as whether a selection branch was considered exhausted (if diversity dropped to only a few predominant sequences or antigen binding did not increase after 2 rounds).

The 2784 individual yeast clones identified from the library selections were individually screened as follows: after each round of selection, individual yeast cells were spotted onto SDCAA agar plates using a FACS-Aria II instrument (BD Biosciences) and screened in the following flow cytometric antigen binding assay: colonies of individual clones were grown until colonies reached 2 mm in diameter and then transferred to a deep-well plate containing 600 μl of liquid SDCAA media. Cultures were grown at 30°C with shaking at 1000 rpm overnight. Expression of the aga2-IgG protein scaffold was induced by replacing growth media with SGRCAA induction media at an optical density of 1 (OD600 = 1). Cells were incubated with either biotinylated recombinant human dimeric antigen or mouse Fc fragment to distinguish yeast clones binding to the Fc portion of recombinant hCD137 antigen. Yeast cells that bound either protein were labeled using streptavidin-APC. FITC-labeled anti-CH2 antibody was also used as a structural marker for IgG. To analyze the screening output, binding to the Fc fragment was plotted and any clone that bound biotinylated Fc was discarded. Binding was defined as more than 0.2% of cells that were positive for APC fluorescence, with a gate set using unstained and negative control samples.

Selections were repeated with varying antigen concentrations and conditions such as increasing induction temperature, decreasing selection stringency, or decreasing the number of rounds to increase the number of candidates. Sequencing of the candidates revealed very low diversity of output products, with only 9 Fcab clones having unique sequences identified: FS22-053, FS22-172, FS22-173, FS22-174, FS22-175, FS22-176, FS22-177, FS22-178, and FS22-179.

Example 3: Characterization of human CD137 Fcab from naive selections

3.1 Obtaining Fcab to human CD137 in the format of a “mock” MAT ²

^MAT2 mimic antibodies were generated consisting of IgG1 molecules containing 76 anti-human CD137 Fcab clones isolated from phage selections and 9 clones isolated from yeast selections to allow characterization of Fcab in the ^MAT2 format. ^MAT2 mimics were generated by replacing a portion of the Fcab CH3 domain containing the AB, CD, and EF loops with the corresponding region of the CH3 domain of the HelD1.3 anti-hen egg lysozyme antibody. The production of HelD1.3 is described in Tello et al. 1993. The heavy and light chain sequences of HelD1.3 are shown in SEQ ID NOs: 186 and 173, respectively. ^MAT2 mimic molecules were generated by transient expression in HEK293-6E cells. To assess the amount of protein produced, IgG protein content was quantified by biolayer interferometry using the Octet QKe platform with PALL Protein A quantification biosensors (18-5021). Proteins were purified by Protein A affinity chromatography using MabSelect SuRe™ columns. 53 phage-derived anti-CD137 MAb² proteins showed measurements below the detection limit and were therefore determined to be unsuitable for further analysis. 32 MAbs were purified using MabSelect SuRe™ Protein A columns (GE Healthcare, 11003494): FS22-005, FS22-007, FS22-033, FS22-042, FS22-049, FS22-050, FS22-052, FS22-053, FS22-054, FS22-167, FS22-169, FS22-170, FS22-171, FS22-172, FS22-173, FS22-174, FS22-175, FS22-176, FS22-177, FS22-178, FS22-179, FS22-180, FS22-181, FS22-183, FS22-184, FS22-186, FS22-187, FS22-191, FS22-192, FS22-193, FS22-194, FS22-195.

In order to compare expression levels, some of the earlier Fcabs were also subcloned and expressed as soluble Fcabs (containing the truncated hinge region) in HEK293-6E cells and purified using MabSelect SuRe™ Protein A columns. Interestingly, some of the Fcabs were found to exhibit significantly better biophysical behavior and production yields in the mock ^MAB2 format than as soluble Fcabs. This was the case for the naive clone FS22-053, which was produced in very low yields as soluble Fcab, but showed a 25-fold improvement when expressed in the mock MAB² format, resulting in a larger number of clones available for characterization.

3.2 Binding to recombinant antigen according to BLI

31 of the purified mAb2 mimic molecules (except clone FS22-175) were tested for binding to recombinant human antigen in a single-point binding experiment by biolayer interferometry using the Octet QKe platform. Streptavidin biosensors for BLI (PALL 18-5021) were used to capture biotinylated hCD137-mFc-Avi antigen at 10 μg/mL in kinetic buffer (PALL). The sensors were then immersed in a well containing purified ^mAb2 diluted 1:1 in the same kinetic buffer for 240 sec, followed by a well containing 1× kinetic buffer for 240 sec. Binding candidates were classified by a simple Boolean yes/no criterion based on the definition of a BLI response that was greater than the response of buffer and the control HelD1.3 wild-type IgG1 mAb (G1/HelD1.3): 12 mAbs did not bind ² , and 19 did bind the CD137-coated sensors (FS22-007, FS22-033, FS22-042, FS22-049, FS22-050, FS22-052, FS22-053, FS22-054, FS22-169, FS22-172, FS22-173, FS22-174, FS22-179, FS22-180, FS22-181, FS22-183, FS22-187, FS22-194, FS22-195).

3.3 Activity of selected mimic mAb ² to CD137 in human NF-κB reporter assay

Multimerization and clustering are required for TNFR signaling (Bitra et al, 2017). CD137 clusters and activates NF-κB signaling upon interaction with its cognate ligand CD137L. Agonist molecules mimic the ligand in triggering CD137 clustering and activation, thereby activating NF-κB signaling. Some agonist antibodies are known to intrinsically induce CD137 clustering upon binding, e.g. urelumab, while others require additional cross-linking of the antibody itself to induce CD137 clustering, e.g. utomilumab (Fisher et al, 2012). Fc-gamma receptors on effector cells are known to induce such cross-linking in vivo, although it is inefficient and may occur away from the site of therapeutic interest. Because various dose-limiting toxicities have been associated with treatment with some anti-CD137 antibodies, it was decided to select CD137-binding Fcabs that are intrinsically incapable of agonist activity, but to select only those that required additional cross-linking to induce CD137 clustering. Thus, an assay was developed that could detect activation of the NF-κB signaling pathway in the cell following clustering of cell-surface expressed CD137 by cross-linked antibodies, but which showed little activity when the antibodies were not cross-linked. This assay was then used to test the agonist functional activity of 27 anti-CD137 Fcab clones in the mock mAb ² format and 6 anti-CD137 Fcab clones in the anti-CD20 mAb² format, regardless of whether the Fcabs were detected binding to the recombinant antigen by BLI or not.

Protein L was used as a cross-linking agent to trigger cross-linking of mock mAb ² by their Fab moieties in the assay, and NF-κB activation was measured.

The cDNA encoding human CD137 (SEQ ID 180) was subcloned into the pMSCV-neomycin vector (Takara Clontech, catalog #634401) using the restriction enzymes EcoRI-HF and XhoI. The RetroPack PT67 cell line (Clontech, catalog #631510) was used to produce retroviral particles according to the manufacturer's protocol. This retrovirus was subsequently used to transduce HEK.FRT.luc cells, which had previously been generated by transducing the Flp-In T-REx 293 HEK cell line (Life Technologies, R780-07) with the Qiagen Cignal Lenti NFkB Reporter (luc) lentivirus (Qiagen, catalog #336851) containing an NF-κB-responsive promoter driving luciferase expression. HEK.FRT.luc.hCD137 cells were used to screen a mimic MAb² containing CD137-binding elements identified in the selections.

A 2 μM dilution of each mock mAb ² was prepared in DFBS (Life Technologies, 14190169) and further diluted 1:3 in reporter cell medium (DMEM (Gibco, catalog #61965-026); 10% fetal bovine serum (FBS) (Gibco, catalog #10270-106); 1× penicillin/streptomycin (Gibco, catalog #15140-122); blasticidin 15 μg/mL (Melford Laboratories Ltd. catalog #B1105); puromycin 5 μg/mL (Life technologies, catalog #A11113803); zeocin 100 μg/mL (InvivGen, catalog #11006-33-0); geneticin 500 μg/mL (Life Technologies, Cat. #10131-027). Protein L (Life Technologies, 21189) was used as an artificial cross-linker and was mixed with MAb² molecules at a molar ratio of 1:4. After 24 h incubation, cells were treated with 100 μL of Promega Bio-Glo™ Luciferase Assay Reagent (Promega, Cat. #G7941) according to the manufacturer's instructions and luminescence was measured with an integration time of 0.5 second on a plate reader with Gen5 software, BioTek. Luminescence values are a measure of luciferase produced in response to activation of the NF-κB signaling pathway by CD137 clustering induced by cross-linked Fcabs. Luminescence values were plotted against log Fcab concentration and the resulting curves were fitted using the Log(agonist) response equation in GraphPad Prism.

Candidates were identified as those that had at least a 10-fold increase in luciferase signal upon cross-linking with protein L compared to no cross-linking. These clones were determined to be able to induce CD137 clustering and further activation of downstream signaling pathways. Of all clones tested, two were able to induce a 10-fold increase in luciferase upon cross-linking, FS22-053 and FS22-172, although an _EC50 could not be determined for either. Both were selected for further characterization in the DO11.10 T cell activation assay. Surprisingly, the remaining clones did not show activity under cross-linking conditions despite binding to the CD137 target by BLI, possibly indicating that they were binding to an off-target epitope on CD137 or that the affinity of such clones was insufficient to bind CD137 strongly enough to initiate the NF-κB signaling cascade.

Overall, although more than 30 Fcabs were tested, only two Fcabs (FS22-053 and FS22-172) were identified from naive selections that displayed target function in the NF-κB reporter assay upon cross-linking and had low activity in the absence of cross-linking.

3.4 Activity of selected mimic mAb ² to CD137 in T cell activation assay DO11.10

Clustering of CD137 by agonist molecules on activated T cells induces T cell activation and subsequent signaling, leading to, but not limited to, IL-2 production. Since FS22-053 and FS22-172 were identified as having activity in the NFKB reporter assay, their ability to activate CD137 was tested in a T cell activation assay. The DO11.10 T cell activation assay was developed using DO11.10 T cells engineered to overexpress human CD137, and T cell activation was assessed by measuring IL-2 release.

DO11.10 T cells (National Jewish Health) were transduced with a lentiviral vector designed to overexpress murine or human CD137 as described previously. In addition to FS22-053 and FS22-172, the following clones were tested in this DO11.10 T cell activation assay: FS22-007, FS22-033, FS22-042, FS22-049, FS22-050, FS22-052, FS22-054 (all in the HelD1.3 mAb ² “mock” format). Dilutions of mAb ² or the positive control mAb, 20H4.9, with or without recombinant protein L as a cross-linker (Life Technologies, 21189) were prepared and added to DO11.10.hCD137 cells in a 96-well round-bottom plate that had been coated overnight with 0.1 μg/ml anti-CD3 antibody (clone 17A2, BioLegend, 100208). After 18 h incubation, supernatants were collected and analyzed with a Mouse IL-2 ELISA kit (eBioscience, 88-7024-86) according to the manufacturer's instructions. Plates were read at 450 nm using a plate reader with Gens software, BioTek. Absorbance values at 570 nm were subtracted from the 450 nm values (correction). The standard curve for calculating cytokine concentration was based on a four-parameter logistic curve fit (Gens Software, BioTek). mIL-2 concentration was plotted against the log concentration of mAb ² or reference mAb, and the resulting curves were fitted using the Log(agonist) response equation in GraphPad Prism.

Clones FS22-053 and FS22-172 showed significantly increased activity when cross-linking protein L in this assay. FS22-053 had 126 nM activity in the absence of cross-linking and 21 nM activity when cross-linked (a 6-fold improvement), while FS22-172 had 950 nM activity in the absence of cross-linking and 44 nM activity when cross-linked (a 22-fold improvement). Based on this, both clones were selected for affinity maturation. Additionally, although clone FS22-033 did not show activity in this assay, it was also selected for affinity maturation because it was thought to be likely to bind the recombinant antigen at a different epitope, as it did not compete with clones FS22-053 and FS22-172 for binding in the BLI binding assay (data not shown). It was thought that improving the binding affinity of the clone to CD137 could also lead to improved functional activity (see Example 4.1).

3.5 Obtaining Fcab to human CD137 in the format MAT ² to CD137/CD20

A panel of anti-CD137 Fcabs (FS22-053, FS22-175, FS22-176, FS22-177, FS22-178, FS22-179) were generated to allow characterization of Fcabs in a more biologically relevant mAb ² format. MAb ^2s were generated by replacing the portion of the Fcab CH3 domain containing the AB, CD, and EF loops with the corresponding region of the CH3 domain of anti-CD20 clone 2F2 (from US 8529902 B2) to generate mAb ^2s that bound both CD137 and CD20. These anti-CD137/CD20 mAb ^2s were produced by transient expression in HEK293-6E cells and purified using MabSelect SuRe™ Protein A columns.

3.6 Activity of MAb ² to CD137/CD20 in the T-cell activation assay DO11.10

Example 3.4 describes a T cell activation assay in which mAb2 mimics to CD137 were cross-linked using the L protein. This configuration provided a robust and reproducible means of screening a large number of molecules, although the higher order structures formed by the antibodies and ^mAb2 upon cross-linking are suboptimal and atypical of the physiological environment. A more biologically relevant configuration is one in which the mAb2 molecule is cross-linked via linkage of its Fab arms to a target present in a biological system. This in vitro cell system optimizes the presentation of antibodies and ^mAb2 by linking the Fab to cell membranes, triggering antibody clustering, which in turn increases the affinity of the Fcab arm to CD137 for its target on T cells.

CD20 ⁺ Daudi cells (ATCC CCL-213) were seeded in a 96-well round-bottom plate at a 1:1 ratio with the DO11.10.hCD137 cells used in Example 3.4. The 6 clones obtained in Example 3.5 in the anti-CD137/CD20 MAb² were tested in this DO11.10 T cell activation assay. Dilutions of MAb ² or positive control MAb were prepared with or without recombinant protein L as a cross-linker (Life Technologies, 21189) and added to DO11.10.hCD137 and Daudi cells in a 96-well round-bottom plate that had been coated overnight with 0.1 μg/ml anti-CD3 antibody (clone 17A2, BioLegend, 100208). After 18 h of incubation, supernatants were collected and analyzed using a mouse IL-2 ELISA kit (eBioscience, 88-7024-86) according to the manufacturer's instructions. Plates were read at 450 nm using a plate reader with Gens software, BioTek. Absorbance values at 570 nm were subtracted from the 450 nm values (Correction). The standard curve for calculating cytokine concentration was based on a four-parameter logistic curve fit (Gens Software, BioTek). mIL-2 concentration was plotted against the log concentration of mAb ² or reference mAb, and the resulting curves were fitted using the Log(agonist) response equation in GraphPad Prism. Of all clones tested, FS22-053 was the only one that showed significant IL-2 production in this CD20 cell-based cross-linking configuration with an _EC50 of 0.3 nM (and 126 nM in the absence of CD20 ⁺ cells).

Examples 3.4 and 3.6 show that FS22-053 was able to trigger CD137 clustering and activation on the surface of DO11.10 T cells when cross-linked by either protein L or cell-based cross-linking via Fab binding to another target in the MAb² formats tested (HelD1.3 and CD20).

Example 4: Affinity maturation of Fcab against a human protein

As discussed previously, three clones were selected for affinity maturation based on their functional characteristics in the NF-κB reporter assay, DO11.10 T cell activation assays (FS22-053 and FS22-172), or those thought to bind to a different region of CD137 (FS22-033).

4.1 Affinity Maturation of FS22-033

Two yeast display libraries and two phage display libraries were constructed from clone FS22-033, one by randomizing five residues in the AB loop of the CH3 domain and the other by randomizing five residues in the EF loop of the CH3 domain using ELLA primers. The ELLA primers define the codons used for each amino acid and their relative abundance in the mixture. Only cysteine was excluded from the mixture, and no other amino acids were biased.

For phage libraries FS22-033 AB and FS22-033 EF, two rounds of selections for affinity matured clones were performed using 200 nM hCD137-mFc-Avi in round 1 and 10 nM hCD137-mFc-Avi in round 2. The 96 clones from the output of each selection were screened by phage ELISA. This screen identified 24 unique clones (FS22-033-001 through FS22-033-024), all of which were generated as mock MAb² containing HelD1.3 and then tested to determine improved binding kinetics by BLI as described in Example 3.2. None of these 24 affinity-matured clones performed better than the original clone FS22-033 in this assay and were therefore not used. For the yeast libraries FS22-033 AB and FS22-033 EF, three rounds of selections were performed as follows: rounds 1 and 2 using 300 nM hCD137-mFc-Avi, then a third round using 300 nM hCD137-mFc-Avi, 300 nM cynomolgus CD137-mFc-Avi, or 300 nM hCD137-Avi-his. A total of 1056 clones from rounds 2 and 3 outputs were sequenced, all identified unique clones were screened in a flow cytometric antigen binding assay to determine improved antigen binding compared to the original clone FS22-033 using 30 nM dimeric recombinant human antigen, and clones were ranked by the percentage of APC ⁺ cells, which correlates with binding strength. The five best clones (FS22-033-025, FS22-033-026, FS22-033-027, FS22-033-028, FS22-033-029) were then prepared as HelD1.3 MAb² (as described in Example 3.1) and binding to dimeric recombinant human antigen was tested by BLI as described previously, but none of the clones showed an improved kinetic profile compared to the original clone FS22-033 and therefore these clones were not used further.

4.2 Affinity Maturation of FS22-053 and FS22-172

Four yeast display libraries were constructed from Fcab clones FS22-053 and FS22-172. Seven residues (at positions 15-16.1 according to IMGT) were randomized using ELLA primers with the same trinucleotide distribution as described in Example 4.1 in the AB loop of the CH3 domain of each clone to generate libraries FS22-053 AB and FS22-172 AB. Five residues (at positions 92-94 and 97-98 according to IMGT) were randomized using ELLA primers in the EF loop of the CH3 domain to generate libraries FS22-053 EF and FS22-172 EF.

For libraries FS22-053 AB and FS22-053 EF, and FS22-172 AB and FS22-172 EF, three or four rounds of selection were performed on the yeast libraries to select for affinity matured clones using either the dimeric hCD137-mFc-Avi antigen or the monomeric hCD137-Avi-His antigen. The monomeric antigen was used in alternation with the dimeric antigens to ensure that clones retained affinity for the antigen and were not binding solely by avidity. The use of monomeric or dimeric antigen, as well as the concentration used, was determined empirically during each round by flow cytometry by determining whether enrichment was observed compared to the monomeric or dimeric antigen in the previous round. Whenever possible, a sorting gate over the parental molecules was used to isolate clones with affinity maturation relative to the parental molecule. The selection pressure was increased to 1 nM dimeric antigen. During each round of selection, individual clones were spotted onto agar plates to assess the progress of selection. Each clone was grown and induced individually, and then its binding and structural parameters were determined by flow cytometry using biotinylated dimeric antigen as well as anti-CH2 structural markers as described previously. This screening cascade was followed to ensure that selection success could be determined based on the selection of clones from the selection output, as well as to allow early screening of individual clones that could subsequently be produced as soluble proteins.

A total of 1152 individual yeast clones were screened for binding to biotinylated recombinant antigen in a flow cytometric antigen binding assay as described previously. Selections from the FS22-053 EF library resulted in the enrichment of 138 unique loop sequences. Similarly, 30 unique loop sequences were isolated from the FS22-172 AB library. The FS22-053 AB and FS22-172 EF libraries did not contain any clones that showed any improvement in binding compared to the parental clones. Sequence analysis among the best binding clones from the FS22-053 EF and FS22-172 AB libraries revealed a conserved PPY sequence pattern in the AB loops. Since this sequence was conserved after affinity maturation and was independently selected in two separate Fcab lines, it may be important for binding to the CD137 epitope. Furthermore, the LE or LD sequence pattern is conserved in the EF loops of the CH3 domain of both FS22-053 and FS22-172 clones, suggesting that this amino acid motif in the EF loop is required for improved binding.

To assess the progress of the selections and the need for recombination of mutated AB and EF loops between affinity matured clones, the top five unique clones from the FS22-053 EF library (FS22-053-008, FS22-053-009, FS22-053-010, FS22-053-011, FS22-053-012), ranked by demonstrating specific binding to 10 nM dimeric human antigen (>30% APC-positive cells in flow cytometric binding assay), and the top six unique clones from the FS22-172 AB library (FS22-172-001, FS22-172-002, FS22-172-003, FS22-172-004, FS22-172-005, FS22-172-006, all showing >10% APC-positive cells when screened using 10 nM dimeric human antigen in the same assay) were prepared as mock mAb ² (HelD1.3) and model mAb ² (PD-L1) mAb² to assess the functional and kinetic improvement of randomized loops.

Example 5: Construction, expression and characterization of an affinity-matured anti-human CD137 Fcab in a “mock” mAb² format

5.1 Construction of Fcab to human CD137 in the format of “mock” and model MAT²

16 affinity matured clones derived from the original clone FS22-053 (FS22-053-001 to FS22-053-016) and 6 clones derived from the original clones FS22-172 (FS22-172-001 to FS22-172-006) were generated in the MAb ² format. Clones FS22-053-001 to FS22-053-007 were not used further because they were not expressed in the MAb ² format at a level that would allow subsequent purification for further testing and characterization. The remaining clones were found to have at least 95% sequence identity in their CH3 domain when compared to the CH3 sequence of the original clone from which they were derived. A percent sequence similarity matrix was generated by comparing the position of each amino acid with the position of the reference sequence (parent clones FS22-053 or FS22-172). (Figure 1D shows the percent sequence identity of the CH3 domain of clones FS22-053-008, FS22-053-016, and FS22-053-017 (see Example 10.1) compared to the parent CH3 domain of FS22-053. Figure 1E shows the percent sequence identity of the CH3 domain of clones FS22-172-001 through FS22-172-006 compared to the parent CH3 domain of FS22-053.

Mock antibodies MAb² containing the human CD137 Fcab in HelD1.3 were generated for further characterization of the affinity matured Fcab in MAb² format. These MAb² were generated as described in Example 3.1.

Model mAbs ² containing Fcab to human CD137 as well as the Fab region binding PD-L1 (clone YW243.55.S70 from US 8217149 B2) were also prepared. They were prepared in a similar manner to that described in Example 3.1 by replacing the CH3 portion of the PD-L1-binding antibody containing the AB, CD and EF loops with the corresponding Fcab region. These model mAbs to PD-L1 contain a LALA mutation in the CH2 domain (AA). Introduction of the LALA mutation in the CH2 domain of human IgG1 is known to reduce Fcγ receptor binding (Bruhns, P., et al. (2009) and Hezareh M., et al. (2001)).

CD137/HelD1.3 and CD137-AA/PD-L1 MAbs were generated by transient expression in HEK293-6E cells and purified using MabSelect SuRe™ Protein A columns.

5.2 Activity of human Fcab in the mimic MAb ² format in a human NF-κB reporter cell assay

The functional activity of the affinity-matured anti-CD137 Fcabs in the mock MAb² format (HelD1.3) listed in Table 3 was tested in the same NF-κB luciferase assay described in Example 3.3. Luminescence was measured with an integration time of 0.5 seconds in a plate reader with Gen5 software, BioTek. The results of this assay are shown in Table 3 and Figure 2. As expected, none of the Fcabs showed activity without cross-linking to the L protein (-XL). All affinity-matured anti-CD137 Fcabs showed a significant improvement over the parental anti-CD137 Fcabs, for which, although positive in this assay, calculation of the _EC50 value was not possible (see Example 3.3). FS22-053-008 and FS22-172-003 showed the best activity of each family with the lowest EC ₅₀ in cross-linking to protein L(+XL).

Table 3

Clone Fcab (in MAT ² HelD1.3 format) NF-kB signaling with or without cross-linking of protein L (XL) -XL +XL (EC ₅₀ , nM) FS22-053-008 Not applicable 26.34 FS22-053-009 Not applicable 64.57 FS22-053-010 Not applicable 47,48 FS22-053-011 Not applicable 32.78 FS22-053-012 Not applicable 119.3 FS22-172-002 Not applicable 124.1 FS22-172-003 Not applicable 32.64 FS22-172-004 Not applicable 123.6 FS22-172-005 Not applicable Not applicable FS22-172-006 Not applicable 382.4

N/A - not applicable because the low signal did not allow EC ₅₀ to be determined.

5.3 Activity of Affinity-Matured Human Fcab in the Model MAb ² Format in the Human T-Cell Activation Assay DO11.10

The functional activity of the affinity-matured human Fcabs in the model MAb² format (PD-L1 LALA) listed in Table 4 was tested in the DO11.10 T cell activation assay, which was similar to the assay described in Example 3.6.

HEK.mPD-L1 cells were generated by subcloning the cDNA encoding mouse PD-L1 (SEQ ID NO: 188) into the pcDNA5FRT vector (Life Technologies) using KpnI and NotI restriction sites and transforming the vectors into the Flp-In T-REx 293 cell line (Life Technologies, R780-07) using Lipofectamine 2000 (Life Technologies, 11668-019). Cells were grown in DMEM containing 10% fetal bovine serum (FBS), 100 μg/ml hygromycin B (Melford Laboratories Ltd, Z2475) and 15 μg/ml blasticidin (Melford Laboratories Ltd, B1105) for 3-4 weeks until colonies of stably transformed cells were formed. These colonies were amplified in the presence of 1 μg/ml doxycycline (Sigma Aldrich, D9891), and PD-L1 expression was tested using PE-conjugated mouse PD-L1 antibody (MIH5) (BD Biosciences, 558091).

Cells were detached using cell dissociation buffer, washed once with PBS, and 2× ¹⁰⁵ cells were seeded into wells of a 96-well plate and then incubated with antibody diluted 1:20 in PBS for 1 hour at 4°C. Cells were washed once in PBS and then measured on an Accuri C6 cytometer (BD Biosciences), and data were analyzed by FlowJoX. Mouse PD-L1 expression was again confirmed.

15 clones obtained in Example 5.1 in the CD137/PD-L1 MAb² were tested in this DO11.10 T cell activation assay. Dilutions of mAb ² or positive control mAb were prepared and added to either DO11.10.hCD137 cells (7.5 × ¹⁰³ cells/well) and HEK.mPD-L1 cells (2 × ¹⁰⁴ cells/well) or to DO11.10.hCD137 cells (7.5 × ¹⁰³ cells/well) and HEK cells that had not been transduced to express mPD-L1 (2 × ¹⁰⁴ cells/well) in a 96-well flat-bottom plate that had been coated overnight with 0.1 μg/ml anti-CD3 antibody (clone 17A2, BioLegend, 100208). After 18 h incubation, supernatants were collected and analyzed with a mouse IL-2 ELISA kit (eBioscience, 88-7024-86) according to the manufacturer's instructions. Plates were read at 450 nm using a plate reader with Gens software, BioTek. Absorbance values at 570 nm were subtracted from the 450 nm values (Correction). The standard curve for calculating cytokine concentration was based on a four-parameter logistic curve fit (Gens Software, BioTek). mIL-2 concentration was plotted against the log concentration of mAb ² or reference mAb, and the resulting curves were fitted using the Log(agonist) response equation in GraphPad Prism. T cell activation was detected by measuring IL-2 release.

The results of this assay are shown in Table 4 and Figure 3. No T cell activity was observed without cross-linking by binding to PD-L1 expressing cells (no cell-based XL bar). After cross-linking, all mAbs ² had strong T cell activity as evidenced by the release of high levels of IL-2 and subnanomolar _EC50 values. With increasing concentrations, the positive control anti-human CD137 mAb, G1-AA/20H4.9, showed an increase in mIL-2 release, however, the maximal release was significantly less than that for anti-human CD137 Fcab. All clones except FS22-053-009 and FS22-172-005 had _EC50s less than 0.3 nM, so were as good or better than the positive control. The lowest observed Emax value, which is a measure of maximal T cell activation and may be related to greater antitumor activity of T cells in vivo, was 7758 pg/mL, which was higher than that of the positive control.

Table 4

Clone Fcab (in the format of the model MAT ² to PD-L1) or MAT IL-2 release with or without mPD-L1 HEK cross-linking No XL Cell Based +XL Emax (IL-2 pg/ml) +XL EC ₅₀ (nM) FS22-053 Not applicable 5366 0.28 FS22-053-008 Not applicable 8176 0.28 FS22-053-009 Not applicable 8083 0.52 FS22-053-010 Not applicable 8301 0.15 FS22-053-011 Not applicable 7923 0.28 FS22-053-012 Not applicable 9808 0.20 FS22-053-013 Not applicable 15551 0.12 FS22-053-014 Not applicable 12849 0.09 FS22-053-015 Not applicable 17079 0.18 FS22-053-016 Not applicable 18003 0.10 FS22-172 Not applicable 5952 0.53 FS22-172-001 Not applicable 10006 0.23 FS22-172-002 Not applicable 7001 0.20 FS22-172-003 Not applicable 10958 0.18 FS22-172-004 Not applicable 7758 0.24 FS22-172-005 Not applicable 13021 1.27 FS22-172-006 Not applicable 9089 0.32 G1-AA/20H4.9 Not applicable 3755 0.26

N/A - not applicable because the low signal did not allow EC ₅₀ to be determined.

5.4 Assay for Activation of Primary Human CD8 ⁺ T Cells

The activity of Fcab to activate CD137 on HEK cells overexpressing CD137 is shown in Example 5.3. To test the activity of Fcab on cells that were not engineered to overexpress CD137, a primary human T cell assay was required. Activated cytotoxic CD8 ⁺ T cells are responsible for the direct killing of cancer cells and express CD137 on their cell surface (Ye et al, 2014). Clustering of CD137 is known to be key for the induction of downstream signaling and further activation of CD8 ⁺ T cells. Therefore, a CD8 ⁺ T cell activation assay was used to assess the ability of Fcab (in the MAb ² format, as detailed below) to trigger clustering and downstream signaling of CD137. CD8 ⁺ T cell activation was determined by IL-2 release.

For T cell isolation, peripheral blood mononuclear cells (PBMCs) were isolated from leukocyte depletion cones, a by-product of platelet donation. Briefly, the contents of the leukocyte depletion cones were washed with PBS and overlaid onto a Ficoll gradient (Sigma-Aldrich, 1440-02). PBMCs were isolated by centrifugation and cells that did not cross the Ficoll gradient were recovered. PBMCs were further washed with PBS and the remaining red blood cells were lysed by adding 10 ml of 1× Red Blood Cell Lysis Buffer (eBioscience, 00-4300-54) according to the manufacturer's instructions. CD8 ⁺ T cells were isolated from PBMCs present in the eluate using the CD8 ⁺ T Cell Isolation Kit II (Miltenyi Biotec Ltd, 130-096-495) according to the manufacturer's instructions.

Incubation with anti-CD3 antibody was used as the first signal to trigger initial T cell activation. 96-well flat-bottomed tissue culture plates were coated with 8 μg/ml anti-CD3 antibody (clone UCHT1, R&D Systems, MAB100-SP) in PBS overnight at 4°C. Plates were then washed 3 times with 200 μl PBS.

For cell-based cross-linking of affinity-matured anti-CD137 human Fcab in the model mAb ² anti-PD-L1 format, HEK293 cells overexpressing hPD-L1 (HEK.hPD-L1) were generated essentially as described in Example 5.3, but by subcloning cDNA encoding human PD-L1 (SEQ ID NO: 187) in place of murine PD-L1. HEK.hPD-L1 cells were seeded at 2× ¹⁰⁵ cells/well in anti-CD3 antibody (8 μg/ml) coated 96-well flat-bottom plates in 100 μl T cell culture medium (RPMI medium (Life Technologies, 61870-044) with 10% FBS (Life Technologies), 1× penicillin/streptomycin (Life Technologies, 15140122), 1 mM sodium pyruvate (Gibco, 11360-070), 10 mM Hepes (Sigma-Aldrich, H0887), 2 mM L-glutamine (Sigma-Aldrich, G7513), and 50 μM 2-mercaptoethanol (Gibco, M6250)). Following attachment of HEK.hPD-L1 cells or HEK cells that were not transduced to express hPD-L1, after 4 hours of incubation, all T cell culture medium was removed and replaced with 100 μl of T cell culture medium containing T cells at a concentration of 5.0× ¹⁰⁵ cells/ml, yielding 5.0× ¹⁰⁴ cells/well.

MAb ² was diluted in T cell medium to 2× final concentration starting at 500 nM and titrated 1:3. 100 µl of MAb ² titration was added to cells to give a total assay volume of 200 µl and 1× antibody concentration.

Each of the positive control anti-CD137 antibody (G1-AA/20H4.9) and negative control isotype IgG antibody (G1-AA/HelD1.3) was diluted in T cell medium to 2× final concentration starting at 500 nM containing 500 nM cross-linker (anti-human CH2 antibody, clone MK1A6 (Jefferis et al., 1985 and Jefferis et al., 1992), prepared in-house) and titrated 1:3. 100 μl of the diluted positive control antibody/cross-linker mixture or negative control IgG antibody/cross-linker mixture was added to the cells to give a total assay volume of 200 μl and 1× antibody concentration.

The assay was incubated at 37°C, 5% _CO2 for 72 h. Supernatants were collected and analyzed using the Human IL-2 Ready-SET-Go! ELISA Kit (eBioscience, catalog #88-7025-88) according to the manufacturer's instructions. Plates were read at 450 nm using a plate reader with Gen5 software, BioTek. Absorbance values at 630 nm were subtracted from absorbance values at 450 nm (Correction). The standard curve for calculating cytokine concentration was based on a four-parameter logistic curve fit (Gen5 Software, BioTek). Human IL-2 (hIL-2) concentration was plotted against log antibody concentration and the resulting curves were fitted using the Log(agonist) response equation in GraphPad Prism.

Table 5 shows the _EC50 values and maximum IL-2 release responses observed in the T cell activation assay in the presence of affinity matured Fcab clones in the model mAb ² format to PD-L1 tested with cell-based cross-linking. The positive control anti-human CD137 mAb, 20H4.9, shows increased hIL-2 release with an _EC50 of 0.5 nM when cross-linked with anti-hCH2 antibody. All clones had activity in the assay, with most showing good activity with subnanomolar _EC50s . MAbs ² containing Fcab FS22-053-007, FS22-053-008, FS22-053-010, FS22-053-011, FS22-053-012, FS22-172-003, FS22-172-004, FS22-172-005 induced the highest T cell responses with the lowest EC ₅₀ values ranging from 0.19 to 0.49 nM. A subset of mAbs ² (containing Fcab FS22-053-008, FS22-053-011, FS22-053-014, FS22-173-003 and FS22-172-004) were also tested without cross-linking PD-L1 expressed on HEK cells and, as expected, showed no activity in this assay. This confirms the activity observed in the NF-kB assay and the DO11.10 T cell activation assay. Figure 4 shows typical IL-2 release curves for the T cell activation assay for MAb ² FS22-053-007-AA/PD-L1, FS22-053-008-AA/PD-L1 and FS22-172-002-AA/PD-L1, FS22-172-003-AA/PD-L1, FS22-172-004-AA/PD-L1.

Table 5

Clone Fcab (in the model format MAT ² to PD-L1) or MAT IL-2 release with hPD-L1 cross-linking by HEK Donor 170505B Emax EC ₅₀ (nM) FS22-053 4344 0.55 FS22-053-007 13119 0.22 FS22-053-008 25112 0.42 FS22-053-009 15230 0.69 FS22-053-010 13978 0.37 FS22-053-011 14959 0.19 FS22-053-012 12282 0.20 FS22-172 10729 1.83 FS22-172-001 19135 0.58 FS22-172-002 20615 2.69 FS22-172-003 30311 0.45 FS22-172-004 22611 0.36 FS22-172-005 23626 0.49 FS22-172-006 27871 0.89 20H4.9 14780 0.89 20H4.9 + XL 44940 0.50

5.5 Determination of Fcab specificity to human CD137 using surface plasmon resonance (SPR)

The specificity of Fcab for human CD137 was tested for human CD137 compared to other related members of the TNFSFR family. 8 of the Fcab were tested in a mock MAb ² (HelD1.3) format and measured by SPR in a Biacore T200 (GE Healthcare) by testing binding to other human TNFRSF receptors: CD40, OX40, and GITR. Amine binding (Amine Binding Kit, GE Healthcare, BR-1000-50) was used to coat human CD40, GITR, and OX40 to approximately 1000 RU on Biacore CM5 chips (GE Healthcare, catalog #29149603). Dilutions of Fcab to human CD137 in mock mAb ² format (FS22-053-008/HelD1.3, FS22-053-009/HelD1.3, FS22-053-010/HelD1.3, FS22-053-011/HelD1.3, FS22-053-012/HelD1.3, FS22-053-014/HelD1.3, FS22-172-003/HelD1.3, FS22-172-004/HelD1.3), starting from 1 μM, were prepared in HBS-EP+ buffer (BR100669) and injected for 3 min at 30 μl/min and then allowed to dissociate in buffer for 4 min. The chip was reconstituted by injecting 10 mM glycine pH 2.5 for 12 s at 30 μl/min. Antibodies specific for different TNFRSF members were used as positive controls to check the coverage of the Biacore chip. The data were subtracted from the two standards and analyzed using BIAevaluation 3.2 software. Fcab did not bind to any of the TNFRSF receptors tested, demonstrating their specificity for CD137. Therefore, Fcab are not expected to cause off-target binding.

5.6 Binding affinity of Fcab for human CD137 in the mock mAb ² format for human, cynomolgus monkey and mouse CD137 according to SPR

The affinity of Fcab to human CD137 (FS22-053-008, FS22-053-011, FS22-053-014, FS22-172-004, FS22-172-004) for human, cynomolgus monkey (cyno), and mouse CD137 was measured by SPR to determine whether the Fcab could be used for testing in animal studies. The anti-human Fab capture antibody was immobilized on all four flow cells of a CM5 S-series flow chip (GE Healthcare, #BR-1005-30) to an average surface density of 6000 RU according to the manufacturer's recommendations (GE Healthcare, Human Fab Capture Kit, #28958325). Each mAb² was captured to approximately 150 RU by injecting 3 μg/mL of mAb² solution diluted in HBS-EP+ buffer (GE Healthcare, #BR1006-69) for 60 seconds at 30 μL/min. Then, various concentrations of human, cynomolgus monkey, or mouse CD137 antigen (non-biotinylated human, cynomolgus monkey, or mouse CD137-mFc-Avi or human CD137-Avi-His) in HBS-EP+ buffer were passed over the chip for 3 minutes at 60 μL/min and then allowed to dissociate for 10 minutes. After each antigen concentration, the chip was reconstituted by injecting 10 mM glycine, pH 2.1, at a flow rate of 30 μL/min for 30 seconds. HBS-EP+ buffer was injected before the highest antigen concentration and after the lowest antigen concentration to subtract the standard, and one randomly chosen concentration was repeated twice. Binding kinetics were approximated using a 1:1 Langmuir model to obtain the equilibrium binding constant (K _D ) for each sample. Data analysis was performed using BiaEvaluation software version 3.2. The results are presented in Table 6.

Analysis of the results revealed improved binding of both human and cynomolgus monkey CD137 by all affinity-matured clones compared to the respective parent molecules. Binding affinities for monomeric human CD137 antigens were weaker (at least 100-fold) than for dimeric human and cynomolgus monkey antigens fused to Fc. As discussed in Example 2, Fcabs were selected to bind preferentially to dimeric CD137 over monomeric forms of CD137, and these data confirm that the selection strategy was successful. This kinetic behavior reduces the likelihood of their binding to monomeric CD137 expressed at minimal levels on unstimulated T cells, resulting in a reduced risk of hepatic or systemic toxicity associated with some anti-CD137 mAb therapies.

The data also show that anti-human CD137 Fcabs bound to dimeric cynomolgus monkey CD137 with an affinity comparable to that for dimeric human CD137.

The ability of Fcab to bind to dimeric mouse CD137 was also tested. None of the clones showed strong binding to mouse antigen (as shown in Table 6, where “not applicable” means that the K _D could not be calculated), except for clone FS22-053-014, which was unexpectedly found to have a K _{D of} 24 nM against mouse antigen. This was unexpected since mouse CD137 and human CD137 share less than 57% sequence homology.

Table 6

Fcab K _D of dimeric human CD137 (nM) K _D of dimeric CD137 of cynomolgus monkey (nM) Fold difference in K _D of monomeric human CD137 relative to K _D of dimeric human CD137 K _D of mouse dimeric CD137 (nM) FS22-053 38 34 Not applicable Not applicable FS22-053-008 4.2 0.9 170 times Not applicable FS22-053-011 5.5 1.3 >200 times Not applicable FS22-053-014 3.2 0.9 100 times 24 FS22-172 52 203 Not applicable Not applicable FS22-172-003 1.5 1.3 >200 times Not applicable FS22-172-004 4.3 3.5 >200 times Not applicable

N/A - not applicable because the low signal did not allow K _D to be determined.

5.7 Determination of Fcab binding valency using heterodimeric and homodimeric Fcabs

Fcabs typically contain two homodimeric Fc chains with antigen-binding sites in the CH3 domains. Because these antigen-binding sites in the two CH3 domains are in close proximity, the binding valence of Fcabs was tested to determine whether the CH3 domains could bind CD137 independently of each other. Heterodimeric Fcabs containing a single CH3 antigen-binding domain were constructed by combining one chain of FS22-172-003 with one wild-type Fc chain using knob-into-hole mutations (knob: T22W, hole: T22S L24A Y66V) (Atwell S et al, 1997). Therefore, each heterodimer contained the CH3 from FS22-172-003 (SEQ ID NO: 139) in one chain and the wild-type CH3 (SEQ ID NO: 4) in the other chain. Fcab was received in MAT ² format.

The heterodimeric Fcab was compared to the homodimeric FS22-172-003 molecule (containing two CH3 antigen-binding domains) using an SPR binding assay. For this experiment, monomeric human CD137-mFc-Avi was preferred over dimeric CD137 antigen. To compensate for the weaker binding to monomeric antigen described in Example 5.6, human CD137-mFc-Avi was immobilized on a CM5 chip at a higher density of 450 RU. Hetero- or homodimeric Fcab were injected and passed over the immobilized antigen. The results in Figure 14 show that heterodimeric Fcab containing only one CD137-binding CH3 domain was able to bind antigen even under these suboptimal conditions. The dissociation rate observed for heterodimeric Fcab was significantly higher than that of homodimeric Fcab. These results confirm that Fcab FS22-172-003 was able to bind to CD137 via one or both CH3 domains and confirmed that the selection strategy described in Example 2 was successful because Fcab was capable of bivalent binding to its target.

5.8 Binding of Fcab to CD137 to cells with different levels of CD137 expression

As described in Example 2, Fcabs were selected for binding CD137 such that they preferentially bind cells with elevated levels of CD137 expression. The bivalent and therefore avid binding of FS22-172-003 was confirmed by SPR in Example 5.6: FS22-172-003 exhibited high avidity and stronger binding to dimeric CD137 compared to monomeric CD137.

A series of DO11.10 cells expressing varying levels of CD137 were prepared as described in Example 3.5. To determine the relative expression of CD137 in each cell line, antibody binding capacity (ABC) was determined according to the manufacturer's protocol (Quantum™ Simply Cellular^®#816 Bangs Labs). Each cell line was ranked in order of CD137 expression levels after background subtraction: hCD137 high (ABC: 1206283), hCD137 intermediate (ABC: 404597), hCD137 intermediate/low (ABC: 143065), hCD137 low (ABC: 14208), hCD137 negative (ABC: 0).

Binding of Fcab to human CD137 in mock MAb² format (FS22-172-003-AA/HelD1.3), positive control antibody (G1-AA/20H4.9), or isotype control antibody (G1-AA/HelD1.3) to each of the cells described above was tested as follows: DO11.10 cells were harvested from T175 cell culture flasks, centrifuged at 1200 rpm for 3 min, and resuspended in ice-cold FACS buffer prepared from DPBS (Life Technologies, 14190169) and 1% BSA (Sigma-Aldrich, A7906) at 2× ¹⁰⁶ cells/mL, and 50 μL per well were seeded in a 96-well V-bottom plate (Costar, 3894). All tested antibodies were diluted in FACS buffer in 120 μl. Then, DO11.10 cells were centrifuged, the supernatant was removed, the cells were resuspended in 100 μl of each antibody dilution and incubated at 4°C for 45 min. The cells were washed twice by centrifugation with 150 μl of FACS buffer, resuspended in 100 μl containing R-Phycoerythrin-conjugated goat anti-human IgG F(ab') ₂ fragment (specific for the γ chain) (Sigma, P8047) diluted 1:1000 in FACS buffer and incubated at 4°C for 45 min. Cells were washed once with 150 µl FACS buffer and then with 150 µl DPBS, resuspended in 150 µl DPBS containing DAPI (Biotium, 40043) at 1:10,000, and read on a BDCantoII or iQue (Intellicyt). Data were analyzed using FlowJo v10 to determine the geometric mean of the live cell PE signal in each well.

As shown in Figure 15, FS22-172-003-AA/HelD1.3 bound more strongly to cells with higher CD137 expression (Figure 15D: CD137int and Figure 15E: CD137high) and did not bind to cells that expressed very low levels of CD137 (Figure 15B: CD137low and Figure 15C: CD137int/low), compared to the positive control G1-AA/20H4.9. In comparison, the positive control G1-AA/20H4.9 bound better to both dimeric and monomeric antigen, with a 10-fold difference in K _D for either dimeric or monomeric antigens, in contrast to FS22-172-003-AA/HelD1.3, which had at least 200-fold weaker binding to monomeric CD137 antigen than to dimeric antigen, as described in Example 5.6.

Example 6: Naive selection of Fcab to mouse CD137

To test the activity of Fcabs against CD137 in mouse models in vivo, Fcabs that specifically bound to mouse CD137 were generated and characterized.

Phage display

Six naive phage libraries displaying the CH3 domain of human IgG1, previously used to select Fcabs binding to human CD137, were used to select Fcabs binding to murine CD137 using recombinant dimeric murine CD137 or cells expressing full-length murine CD137 as antigens.

In-house produced mCD137-mFc-Avi antigen and DO11.10 cells expressing mCD137 (DO11.10.mCD137) were used in selections using six phage libraries. A simple selection scheme was followed in which all rounds of selection (three in total) were performed using 100 nM biotinylated antigen, with a deselection step using 500 nM unlabeled recombinant human Fc fragment. Binders were captured using magnetic beads coated with either streptavidin or neutravidin. In addition, the output products from round 1 that bound to the recombinant antigen were also used in selections on DO11.10.mCD137 cells expressing murine CD137. Briefly, phage outputs were incubated with mouse CD137-null DO11.10 cells to remove unwanted binding elements, such as those that bind non-specifically to the cells. Phage were then incubated with 1× ¹⁰⁷ DO11.10.mCD137 cells. Binders were then eluted by trypsin digestion and subsequently expanded for a second round of selection. Round 3 was performed in a similar manner, increasing the selection pressure by reducing the number of DO11.10.mCD137 cells to 5× ¹⁰⁶ .

All outputs from round 3 recombinant antigen selection (576 clones) and all outputs from round 3 cell selection (576 clones) were screened by phage ELISA (as described previously) and for cellular binding to DO11.10.mCD137 cells. In ELISA, the majority of clones showed high signal intensity for antigen binding (OD450>1). Thus, clones that showed less than 10-fold increase in antigen binding compared to mouse Fc binding were removed. For cell binding, a FITC MFI greater than 5× ¹⁰⁵ was considered positive. Three of the phage libraries performed significantly worse, with many clones showing non-specific binding to DO11.10.mCD137 cells and recombinant antigens. 34 Fcab candidate clones were subcloned and generated as MAb² HelD1.3 as described in Example 3.1.

Yeast display

Four naive yeast libraries displaying the CH1 to CH3 domains of human IgG1, previously used to select Fcabs binding to human CD137, were used to select Fcabs binding to mouse CD137.

A total of 53 separate rounds of selections were performed to identify mouse CD137 binders. Recombinant dimeric biotinylated mouse CD137 antigen (mCD137-mFc-Avi), produced in-house, was used to select binders from naive yeast libraries. Briefly, binders from round 1 were selected by incubating naive libraries with 300 nM recombinant antigen and deselected with 2.5 μM unlabeled mouse IgG2a Fc fragment. The output products were separated using MACS and streptavidin magnetic beads. Three rounds of FACS selections were performed using 300 nM recombinant antigen and 1.5 μM mouse Fc, which were used for selections and deselections, respectively, as described previously in Example 2.

Individual clones from each of rounds 2, 3, and 4 were spotted onto agar plates. To determine the diversity of output products, at least 96 clones from the output of each selection were sequenced. 126 unique clones (50 clones from round 3 for one of the libraries, 48 clones from round 3 for another library, and 18 clones from round 4 for the remaining library) were screened for binding to the recombinant antigen by flow cytometry. Clones that showed more than 10% positive cells in the APC fluorescence channel when incubated with the recombinant antigen and less than 0.2% when incubated with the recombinant mFc were considered candidates.

Example 7: Characterization of Fcab to CD137 from mouse naive selections

7.1 Determination of Fcab specificity to mouse CD137 using BLI

The specificity of Fcab to mouse CD137 for mouse CD137 was tested in the HelD1.3 mock mAb ² format and measured by BLI in the Octet QKe system by testing Fcab binding to other mouse TNFRSF receptors (CD40, OX40, GITR). Streptavidin biosensors (PALL ForteBio 18-5021) were coated with 10 ng/µl mouse CD40, GITR, OX40 receptors (all from R&D Systems and biotinylated using the EZ-Link Sulfo-NHS-SS-Biotin kit from Thermoscientific #21328). Fcab to mouse CD137 in the mock mAb ² format was diluted 1:1 in kinetic buffer (PALL 18-1092) to a final concentration of at least 1 µM. Antigen-coated sensors were immersed in MAb² solutions for 180 sec and then in 1× kinetic buffer for 180 sec. Antibodies to each of the TNFRSF receptors were used as positive controls. Fcab clones FS22m-055, FS22m-063, FS22m-066, FS22m-075, FS22m-135, FS22m-055, FS22m-063, FS22m-066 did not bind to any of the TNFRSF receptors tested, demonstrating their specificity for mouse CD137.

7.2 Activity of mouse Fcab in the mock mAb ² format in a mouse NF-κB reporter cell assay

HEK.FRT.luc cells expressing the mouse CD137 sequence (SEQ ID NO: 184) were prepared following the same methodology as described previously in Example 3.3. MAb2 containing the previously selected anti-mouse CD137 Fcabs were screened using the cell line, HEK.FRT.luc.mCD137, according to the method described in Example 3.3. 56 ^MAb2 were tested, 29 of which were positive for NF-kB activity. Lob12.3 containing human IgG1 Fc with the LALA mutation (G1AA/Lob12.3) was used as a positive control anti-mouse CD137 MAb and showed an increase in luminescence, confirming the correctness of the assay. HelD1.3, also containing a human IgG1 Fc with the LALA mutation, was used as a negative human IgG isotype control to exclude interference from mock human IgG Fab in the assay. Where possible, _EC50 were calculated and ^MAb2s whose activity did not reach a plateau were discarded in favor of ^MAb2s that showed classical sigmoidal activity kinetics. ^MAb2s were ranked in order of _EC50 and fold change in activity after cross-linking to protein L. FS22m-063 was selected based on having the best _EC50 after cross-linking (1.44 nM) and the highest fold change in activity after cross-linking (27-fold). Figure 5 shows that the mouse anti-CD137 Fcab FS22m-063 in the HelD1.3 mock mAb ² format triggers CD137 clustering and NF-kB signaling when cross-linked to the L protein in a HEK mCD137 NF-kB reporter assay.

Example 8: Antitumor activity of Fcab FS22m-063 (in mAb ² FS22m-063-AA/PD-L1) in vivo

After Fcab FS22m-063 was shown to be able to trigger CD137 clustering and activation in vitro, it was desirable to test its ability to activate CD137 in vivo.

8.1 Preparation of Fcab FS22m-063 in MAT ² format for in vivo testing in mice

MAb ² containing Fcab to mouse CD137, FS22m-063 and the PD-L1-specific Fab region was generated using a similar methodology as for the model MAb ² generated in Example 7.1 and tested for in vivo antitumor activity in a syngeneic MC38 mouse tumor model.

Controls: G1-AA/Lob12.3, G1-AA/S70, G1-AA/4420.

Control antibodies for in vivo experiments were generated by fusion of the heavy chain variable region of the anti-PD-L1 antibody, S70 (clone YW243.55.S70 from US 8217149 B2), to the human IgG1 constant region (G1m17) containing the LALA mutation, and the light chain variable region from S70 was fused to the human constant region (Lm1) via the human kappa J region. MAb ² was generated by replacing the CH3 domain of the reformatted construct described above, FS22m-063, and designated "FS22m-063-AA/S70".

8.2 Activity of MAb ² FS22m-063-AA/S70 in the syngeneic MC38 tumor model

Syngeneic mouse models are recognized as suitable mouse systems for testing the antitumor effect of inhibiting therapeutic targets and have been widely used to validate the development of human therapeutics. The syngeneic MC38 tumor model was used in this experiment because MC38 tumors are known to be highly immunogenic, respond to anti-CD137 antibody monotherapy (Kocak et al, 2006), and express PD-L1 (Juneja et al, 2017).

Each female C57BL/6 mouse (The Jackson Laboratory), 9–10 weeks old and weighing 17.92–23.89 g, was rested for 1 week before study initiation. All animals were microchipped and assigned a unique identifier. There were 12 mice in each cohort. The MC38 colon carcinoma cell line (National Cancer Institute, USA) was initially expanded, maintained, and then prescreened for pathogens and shown to be pathogen negative. Each animal received 1× ¹⁰⁶ cells injected subcutaneously into the right flank in 100 μl of serum-free culture medium (Dulbecco's modified Eagle's medium). Mice that were tumor-free at this time point were excluded from the study 7 days after tumor cell inoculation.

MAb ² FS22m-063-AA/S70 and control antibodies (G1-AA/Lob12.3 (CD137 positive control), G1-AA/S70 (PD-L1 positive control), G1-AA/4420 (isotype control)) were administered to mice by intraperitoneal injection at a fixed concentration of 20 μg per dose in DFBS + 1 mM arginine + 0.05 Tween 80. Each mouse received MAb ² or control antibody by intraperitoneal (i.p.) injection of 200 μl on days 7, 9, and 11 after tumor inoculation. Accurate tumor measurements were made, any drug dosing scheduled for the day in question was performed, and mice were closely monitored until the end of the study. Tumor volume measurements were performed using calipers to determine the longest axis and the shortest axis of the tumor. The following formula was used to calculate tumor volume:

LX( ^S2 )/2

Where L = longest axis; S = shortest axis

As shown in Figure 6, mAb ² FS22m-063-AA/S70 demonstrated significant inhibition of tumor growth compared to mice receiving either control antibody. Statistical significance was shown pairwise for growth rates over the entire study duration using a mixed model analysis comparing all groups. As shown in Table 7, surprisingly, all mice receiving mAb ² FS22m-063-AA/S70 were tumor-free at the end of the study, compared to only 4 of 12 mice receiving the combination of anti-PD-L1 and anti-CD137 (G1-AA/S70 + G1-AA/Lob12.3) or anti-PD-L1 or anti-CD137 alone.

Table 7: Tumor-free mice per treatment group in the syngeneic MC38 tumor model

Group Tumor-free mice (%) Number of tumor-free mice G1-AA/Lob12.3 16% 2/12 G1-AA/4420 0% 0/12 G1-AA/S70 16% 2/12 G1-AA/S70 + G1-AA/Lob12.3 33% 4/12 FS22m-063-AA/S70 100% 12/12

The study shows that in mice with a fully functioning immune system, CD137 agonism, presumably through cross-linking via PD-L1, results in reduced tumor growth, presumably through increased cytotoxic activity of CD8 ⁺ T cells in the tumor.

Example 9: Antitumor activity of Fcab FS22m-063 (in mAb ² FS22m-063-AA/PD-1) in vivo

After Fcab FS22m-063 was shown to be able to trigger CD137 clustering and activation in vitro, it was desirable to test its ability to activate CD137 via another Fab target in vivo, in this case a target that is found only on immune cells, PD-1.

9.1 Preparation of Fcab FS22m-063 in MAT ² format for in vivo testing in mice

MAb ² containing Fcab to mouse CD137, FS22m-063 and the PD-1 specific Fab region was prepared using the same methodology as for the model MAb ² prepared in Example 7.1 and tested for in vivo antitumor activity in a syngeneic MC38 mouse tumor model.

Control antibodies (G1/Lob12.3, G1-AA/F2, G1-AA/4420) for in vivo experiments were prepared by fusion of the heavy chain variable region of the anti-PD-1 antibody, F2 (clone PD1-F2 from WO 2004/056875 A1), to the human IgG1 constant region (G1m17) containing the LALA mutation, and the light chain variable region of the F2 antibody was fused to the human constant region (Lm1) via the human kappa J region. MAb ² was prepared by replacing the CH3 domain of the reformatted construct described above, FS22m-063, and designated "FS22m-063-AA/F2".

9.2 Activity of MAb ² FS22m-063-AA/F2 in the syngeneic MC38 tumor model

The syngeneic MC38 tumor model was used in this experiment as described in Example 8.2, with the following exceptions:

Female C57BL/6 mice (Charles River) aged 9–11 weeks were microchipped and assigned a unique identifier. There were 12 mice in each cohort. Each animal received 1× ¹⁰⁶ MC38 colon carcinoma cells injected subcutaneously into the right dorsal flank in 100 μl serum-free medium.

MAb ² FS22m-063-AA/F2 and control antibodies (G1-AA/Lob12.3 (CD137 positive control), G1-AA/F2 (PD-1 positive control), G1-AA/4420 (isotype control) were administered to mice by intraperitoneal injection at a fixed concentration of 20 μg per dose in DFBS + 1 mM arginine + 0.05 Tween 80. Each mouse received MAb ² or control antibody by intraperitoneal (i.p.) injection of 200 μl once the tumor volume reached 50-60 ^mm3 (day 0) and on days 2 and 4 after the first dose. Tumors were measured as described in Example 8.2.

As shown in Figure 7 A and B, MAT²FS22m-063-AA/F2 showed highly significant inhibition of tumor growth compared to mice treated with isotype control, PD-1 antibody as a positive control. As shown in Table 8, surprisingly, 10 of 12 mAb-treated mice²FS22m-063-AA/F2 mice were tumor-free at the end of the study, compared with only 7 of 12 mice treated with the combination of anti-PD-1 antibody and anti-CD137 antibody (G1-AA/F2 + G1-AA/Lob12.3).

Table 8

Group Tumor-free mice (%) Number of tumor-free mice G1/Lob12.3 50% 6/12 G1-AA/4420 0% 0/12 G1-AA/F2 16% 2/12 G1-AA/F2 + G1-AA/Lob12.3 50% 6/12 FS22m-063-AA/F2 83% 10/12

The study shows that in mice with a fully functioning immune system, CD137 agonism, presumably through cross-linking via PD-1 binding, with additional PD-1 blockade, results in reduced tumor growth, presumably through increased cytotoxic activity of CD8 ⁺ T cells in the tumor.

It has also been shown that Fcabs to CD137 can be cross-linked in a MAb ² format by binding the Fab arms to the immune cell target, resulting in clustering and activation of CD137.

Example 10: Antitumor activity of mAb ² to mCD137/MSLN in vivo

Following the demonstration of the efficacy of the MAb² containing Fcab FS22m-063 and Fab to PD-L1 (Example 8), it was desired to test the ability of Fcab to CD137 in the MAb² format to activate CD137 by cross-linking MAb ² via binding of its Fab arms to a tumor-associated antigen (TAA), in this case mesothelin (MSLN). This tumor-targeted approach is expected to benefit from localizing T cell activation to the tumor microenvironment, as MAb ² cross-linking and hence CD137 agonism will only occur at sites where MSLN is expressed.

A syngeneic tumor model expressing murine MSLN was constructed in mice. CT26 colon carcinoma cells (ATCC, CRL-2638) expressing full-length murine mesothelin (SEQ ID NO: 189) were generated by lipofection (Lipofectamine 3000, Thermo Fisher Scientific, catalog # L3000008) using the pcDNA3.1(+) vector (Thermo Fisher Scientific, catalog # V79020). According to the manufacturer's protocol, CT26 cells were transfected with pcDNA3.1 vectors containing murine MSLN cDNA. Stable transfection was achieved using geneticin as a selective antibiotic (at 600 μg/ml) in complete media (RPMI, 10% FBS).

Expression of mouse MSLN on CT26 cells was confirmed by flow cytometry using the MOR6626 antibody as a positive control (WO 2009/068204 A1). Specifically, cells were incubated with the positive control antibody for 1 hour, followed by the use of a fluorescently labeled anti-human IgG detection antibody (Stratech Scientific Ltd, Cat# 109-546-098-JIR) to detect cell binding. Clonal populations were expanded and subsequently analyzed to determine relative expression levels using the same flow cytometry procedure, and a single clone was selected and named CT26.G10.

CT26.G10 tumor growth was confirmed in vivo. Female Balb/c mice (Charles River) aged 8–10 weeks were microchipped and assigned a unique identifier. There were 17 mice per cohort and each animal received 1× ¹⁰⁵ cells injected subcutaneously into the left dorsal aspect in 100 μl of serum-free medium. Tumor volume measurements were made three times weekly using calipers as described in Example 8. The study was performed in accordance with UK Home Office guidelines as described in Example 8.

At the end of the study, tissues were collected and membrane-bound mesothelin expression was confirmed by immunohistochemical staining in formalin-fixed paraffin-embedded (FFPE) tumor tissues as follows: 4 μm FFPE tissue sections were deparaffinized and antigen was retrieval using low pH=6.1 at 97°C (Dako PT Link) followed by peroxidase blocking and protein blocking before incubation with anti-mesothelin primary antibody (LifeSpan Biosciences, cat.# LS-C407883) at 1 μg/mL. Anti-mesothelin antibody was detected using labeled polymer-HRP-conjugated anti-rabbit Ig secondary reagent and DAB (3,3'-diaminobenzidine) chromogenic endpoint (Dako EnVision+ System).

To evaluate the efficacy of Fcab FS22m-063, the following molecules or combinations were tested in vivo: anti-MSLN antibody FS28m-228-010 as a human IgG1 isotype with LALA mutations (G1-AA/FS28m-228-010), Fcab in two formats of “mimicking” mAb ² (FS22m-063-AA/HelD1.3 and FS22m-063-AA/4420), a combination of antibody FS28m-228-010 with a mimicking anti-CD137 mAb ² with LALA mutations (G1-AA/FS28m-228-010 + FS22m-063-AA/HelD1.3), an antibody for the human isotype control (G1-AA/HelD1.3) and finally mAb² to CD137/MSLN (FS22m-063-AA/FS28m-228-010) with LALA mutations.

Each female Balb/c mouse (Charles River), 8–10 weeks old and weighing 20–25 g, was acclimated for one week prior to study initiation. All animals were microchipped and assigned a unique identifier. With the exception of FS22m-063-AA/4420 (n=10 mice), each cohort contained 20 mice. The CT26.G10 colon carcinoma cell line was expanded and cell banks were established. Each animal received 1× ¹⁰⁵ cells injected subcutaneously into the left flank in 100 μl of serum-free medium. Any mice that were tumor-free 12 days after tumor cell inoculation were excluded from the study.

A 200 μg dose of each antibody (~10 mg/kg) was prepared and administered to mice via intraperitoneal (i.p.) injection. Additionally, both FS22m-063-AA/HelD1.3 and G1-AA/FS28m-228-010 were each prepared at 200 μg per dose (~10 mg/kg) for the combination group. 200 μL of the dose was administered to mice on days 12, 14, and 16 (q2dx3) post tumor inoculation. Tumor volume measurements were performed three times weekly using calipers and mice were closely monitored. The study endpoint was determined by humane endpoints based on tumor volume and mouse condition.

As shown in Figure 9, mAb ² FS22m-063-AA/FS28m-228-010 significantly inhibited tumor growth compared to the G1-AA/HelD1.3 isotype control. Table 9 shows a pairwise comparison of tumor growth rates for all treatment groups over the course of the study using a mixed model analysis in which all groups were compared to the G1-AA/HelD1.3 isotype control.

Table 9: Results of pairwise comparisons of tumor growth using a mixed model analysis in which all groups were compared with the G1-AA/HelD1.3 isotype control.

Groups P-values
Mixed model analysis G1-AA/HelD1.3 10 mg/kg (negative control) G1-AA/FS28m-228-010 10 mg/kg 0.4367 Unreliable FS22m-063-AA/HelD1.3 10 mg/kg 0,0017 *** FS22m-063-AA/4420 10 mg/kg 0.7067 Unreliable G1-AA/FS28m-228-010 10 mg/kg
+ FS22m-063-AA/HelD1.3 10 mg/kg 0.2093 Unreliable FS22m-063-AA/FS28m-228-010 10 mg/kg 0,0000 ****

Not significant p≥0.05; *p<0.05; **p<0.01; ***p<0.001; ****p <0.0001

All animals bearing tumors measuring 62.5 ^mm3 or less at the end of the study were scored as complete responders (see Table 10). Thirty-five percent of animals receiving anti-CD137/MSLN mAb ² were complete responders at the end of the study, compared with 0% in the isotype controls G1-AA/HelD1.3, FS22m-063-AA/HelD1.3, FS22m-063-AA/4420, and the combination of FS22m-063-AA/HelD1.3 and G1-AA/FS28m-228-010.

Table 10: Number and percentage of tumor-free mice (tumors≤62 ^mm3 ) at the end of the study in the syngeneic CT26.G10 tumor model.

Groups Mice were tumor-free at the end of the study. G1-AA/HelD1.3 10 mg/kg (negative control) 0/20 (0%) G1-AA/FS28m-228-010 10 mg/kg 1/20 (5%) FS22m-063-AA/HelD1.3 10 mg/kg 0/20 (0%) FS22m-063-AA/4420 10 mg/kg 0/10 (0%) G1-AA/FS28m-228-010 10 mg/kg + FS22m-063-AA/HelD1.3 10 mg/kg 0/20 (0%) FS22m-063-AA/FS28m-228-010 10 mg/kg 7/20 (35%)

Survival analysis (Figure 10 and Table 11) showed that mAb ² FS22m-063-AA/FS28m-228-010 induced a significant survival benefit compared to the G1-AA/HelD1.3 antibody, while the components (G1-AA/FS28m-228-010, FS22m-063-AA/HelD1.3), or G1-AA/FS28m-228-010 + FS22m-063-AA/4420 did not show a survival benefit. Furthermore, mAb ² FS22m-063-AA/FS28m-228-010 resulted in an improved median survival of 42.5 days compared with G1-AA/HelD1.3 (29 days), FS22m-063-AA/HelD1.3 (30 days), FS22m-063-AA/4420 (29 days), G1-AA/FS28m-228-010 (30 days), and the combination of FS22m-063-AA/HelD1.3 with G1-AA/FS28m-228-010 (29 days).

Table 11: Median survival times for animals treated with each compound and results of pairwise statistical analyses (log-rank) in the syngeneic CT26.G10 tumor model.

Groups Median survival (days) P-values
log rank G1-AA/HelD1.3 10 mg/kg 29 G1-AA/FS28m-228-010 10 mg/kg 30 0.993 Unreliable FS22m-063-AA/HelD1.3 10 mg/kg 30 0.3952 Unreliable FS22m-063-AA/4420 10 mg/kg 29 0.9645 Unreliable G1-AA/FS28m-228-010 10 mg/kg + FS22m-063-AA/HelD1.3 10 mg/kg 29 0.4706 Unreliable FS22m-063-AA/FS28m-228-010 10 mg/kg 42.5 <0.0001 ****

Not significant p≥0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

These data demonstrate that cross-linking of MAb ² by MSLN is capable of triggering CD137 agonism in the tumor; the bispecific antibody, which is superior to targeting CD137 and/or MSLN alone (or even in combination), results in significantly improved survival of tumor-bearing mice. Fcab in mock MAb² format FS22m-063-AA/HelD1.3 or FS22m-063-AA/4420, without any Fab targeting MSLN, showed no activity in this study.

Example 11: Selections to obtain Fcab capable of binding to mouse and human CD137

Since it was unexpectedly found that some of the Fcabs that bind to human CD137 also bound to mouse CD137 (see Example 5.5 for binding specificity to human, mouse and cynomolgus monkey CD137), it was decided to determine whether these clones could be improved.

11.1: Site-directed modification to eliminate potential sequence vulnerability in clone FS22-053-014

Clone FS22-053-014 was selected as being cross-reactive between mouse and human because it was shown to bind to the dimeric mouse CD137 antigen by SPR (see Example 5.5). However, upon further sequence analysis, it had a potential sequence vulnerability that could result in post-translational isomerization of the aspartate in the EF loop of its CH3 domain by a Q98D mutation together with the wild-type G99 position to yield a DG motif. Other affinity-matured clones in the FS22-053 lineage (see Table 12) instead contained a Q98E modification at the same position in the EF loop. Q98D was mutated to Q98E by site-directed mutagenesis using the QuickChange II Mutagenesis Kit (Agilent, catalog #200523) according to the manufacturer's recommendations to generate clone FS22-053-017. Table 12 below shows the common LE motif in the EF loop of the CH3 domains of clones FS22-053-008, FS22-172-003, and FS22-172-004, and the DG motif in the EF loop of the CH3 domain of clone FS22-053-014.

Table 12

Loop EF (92-101) G1 CH3 D K S R W Q Q G N V FS22-053-008 D Y W R W L E G N V FS22-053-011 D Y W R W T D G N V FS22-053-014 Y H W R W L D G N V FS22-172-003 G A D R W L E G N V FS22-172-004 G A D R W L E G N V FS22-053-017 Y H W R W L E G N V

Example 12: Characterization of mutant clone Fcab FS22-053-017

12.1 Activity of Fcab FS22-053-017 in Mock MAb ² Format in the Activation Assay of Human CD137-Expressing T Cells DO11.10

Fcab clone FS22-053-017 was subcloned and expressed as a “mock” HelD1.3 MAb² and then compared with clone FS22-053-014, also in the HelD1.3 MAb² format, in the DO11.10 human CD137-expressing T cell activation assay described in Example 3.4. G1-AA/20H4.9 was used as a positive control against CD137 and G1-AA/D1.3 was used as an IgG control. MAb² were tested either without cross-linking to protein L or cross-linked to protein L at a ratio of 1:4.

Table 13

Clone Fcab (in MAT² HelD1.3 format) or MAT Release of IL-2 with or without cross-linking of protein L Without XL +XL Emax (IL-2 pg/ml) +XL EC ₅₀ (nM) FS22-053-014 Not applicable 4174 0.73 FS22-053-017 Not applicable 4956 0.82 G1-AA/20H4.9 Didn't measure 5018 0.82

N/A - not applicable because the low signal did not allow EC ₅₀ to be determined.

The results in Table 13 and Figure 12 show that both Fcab FS22-053-17 in Mock MAb ² format and Fcab FS22-053-014 in Mock MAb ² format had comparable cross-linking activity with protein L in the same DO11.10 T cell activation assay. Thus, the mutagenesis performed did not adversely affect the functional activity. Both clones in Mock MAb ² format had no activity without cross-linking.

As expected, the CD137 positive control had activity only upon cross-linking, and the IgG control had no activity regardless of cross-linking.

12.2 Activity of mutant Fcab FS22-053-017 in mock mAb ² format in the CD137-expressing murine T cell activation assay DO11.10

Clone FS22-053-017 (in the HelD1.3 mAb ² mock format) was also compared with the murine CD137 binding Fcab clone FS22m-063 (also in the HelD1.3 mAb ² mock format) as well as with the original clone FS22-053-014 (in the HelD1.3 mAb ² mock format) in the DO11.10 murine CD137-expressing T cell activation assay described in Example 3.4. The mAb2 molecules were cross-linked to protein L at a molar ratio of 4:1 (mAb ² :protein L).

As expected, all of the molecules tested showed activity as measured by IL-2 release when cross-linked to the L protein, but no activity without cross-linking. FS22m-063, which was selected to bind to mouse CD137, had the best activity in the assay with an _EC50 of 0.39 nM when cross-linked. Both FS22-053-14 and FS22-053-017 had activity in the assay, indicating that function was not lost due to mutagenesis, although FS22-053-017 had a slight loss of activity with an _EC50 that was approximately 8-fold worse than the _EC50 of FS22-053-14 when cross-linked with protein L. Figure 12 shows that the affinity-matured, mouse-human cross-reactive anti-CD137 Fcab FS22-053-014 and FS22-053-017, as well as the mouse anti-CD137 Fcab FS22m-063, in a HelD1.3 mAb ² mock format, activate CD137 when cross-linked with protein L, resulting in the release of mIL-2 in T-cell activation assay DO11.10.

Table 14

Clone Fcab (in MAT ² HelD1.3 format) or MAT Release of IL-2 with or without cross-linking of protein L Without XL +XL Emax (IL-2 pg/ml) +XL EK50 (nM) FS22-053-014 Not applicable 28071 2.04 FS22-053-017 Not applicable 41042 16.74 FS22m-063 Not applicable 24175 0.39 G1-AA/Lob12.3 + protein L Not applicable 21332 2.26

N/A - not applicable because the low signal did not allow EC ₅₀ to be determined.

12.3 Kinetics of FS22-053-017 binding

The equilibrium dissociation constant (K _D ) of Fcab clone FS22-053-017 was compared with that of clone FS22-053-014 by SPR on a Biacore T200 system. For the hCD137-mFc-Avi antigen, the following method was used: anti-human Fab molecule was immobilized on a CM5 chip to an area density of 9,000 to 11,000 response units (RU). Antibodies were diluted to 4 μg/mL in HBS-EP buffer and captured on the anti-Fab molecule at a flow rate of 30 μL/s. Eight different concentrations of hCD137-mFc antigen were used: 200 nM; 66.67 nM; 22.22 nM (included twice), 7.41 nM; 2.47 nM; 0.82 nM, 0.27 nM: 0.091 (diluted in HBS-EP+ buffer) were passed over the captured MAb².

To determine the binding kinetics of mCD137-mFc-Avi, a different approach was used. Murine PD-L1-mFc-Avi was immobilized on a CM5 chip at a surface density of 200 RU. MAbs ² containing Fcab FS22-053-014 and FS22-053-017 in combination with Fab to murine PD-L1 (S70) were diluted to 7.5 μg/mL in HBS-EP buffer and captured with immobilized mPD-L1 protein at a flow rate of 60 μL/s. Eight different concentrations of mCD137-mFc antigen were used: 600 nM; 200 nM; 66.67 nM (included twice), 22.2 nM; 7.1 nM; 2.47 nM; 0.82 nM (diluted in HBS-EP+ buffer) was passed over the captured ^MAb2 .

Data analysis was performed using the Biacore T200 evaluation program. The curves were blank subtracted and fitted using a 1:1 Langmuir binding model with mass transfer, setting RI to zero as a constant and Rmax as a local value. The results are summarized in Table 15 and show kinetic profiles very similar to human antigen for both Fcab clones, indicating that the Q98D to Q98E mutation had no negative effect on binding to human antigen. The 2-fold reduction in binding strength to mouse antigen is also consistent with the functional data reported in Example 11.2.

Table 15

Molecule k _a (1/Ms) k _d (1/s) K _D (nM) FS22-053-014/HelD1.3 8,304×10 ⁴ 3,656×10 ^-4 4,403 FS22-053-017/HelD1.3 8,853×10 ⁴ 4,062×10 ^-4 4,588 FS22-053-014/mPD-L1 1,509×10 ⁵ 4,025× ^-3 27 FS22-053-017/mPD-L1 1,064×10 ⁵ 5,186× ^-3 49

Example 13: Involvement of PPY in antigen binding (alanine scanning of the conserved PPY motif)

As described in Example 5, a PPY sequence motif was identified in the AB loop of two independently selected Fcabs, and this motif was conserved in all affinity-matured clones analyzed. It was therefore desirable to understand the involvement of this motif in CD137 binding and the overall structure of the protein. Alanine scanning is a well-established biological technique used to determine the importance of a particular residue in protein-protein interactions. In general, each of the three amino acids was replaced with alanine residues in turn and then collectively. Alanine is considered the most chemically inert, non-bulky residue and therefore is considered unlikely to contribute to binding.

13.1 Generation of mutant clones for alanine scanning

Mutant clones were generated by site-directed mutagenesis on the parent clones (FS22-172-003 and FS22-053-008). The positions of the amino acid substitutions in the AB loop are summarized in Table 16, all other residues are maintained relative to their corresponding parent residue. The MAb² variants were then transiently expressed in HEK293-6E cells and purified using MabSelect SuRe™ Protein A columns as described in Example 3. The expression yield of the mutants was similar to that of the parent clones, thus indicating that the alanine substitutions did not affect protein production.

Table 16

Original clone Initial AB loop sequence Mutated AB loop sequence Clone name FS22-53-008 NPPYLFS
(SEQ ID NO: 19) NAPYLFS FS22-053-008_APY NPAYLFS FS22-053-008_PAY NPPALFS FS22-053-008_PPA NAAALFS FS22-053-008_AAA FS22-172-003 PYIIPPY
(SEQ ID NO: 138) PYIIAPY FS22-172-003_APY PYIIPAY FS22-172-003_PAY PYIIPPA FS22-172-003_PPA PYIIAAA FS22-172-003_AAA

13.2 Binding kinetics of mutant clones

The binding kinetics of the 2 parental clones and 8 mutant clones were compared using the Octet QK ^e system from ForteBio. Dimeric biotinylated hCD137-mFc-Avi was captured onto streptavidin sensors at 10 μg/mL and then analyzed for interaction with each mAb² at a concentration of 200 nM. Binding of MAb² to the dimeric antigen was impaired for all mutant clones of FS22-172-003 compared to the parental clone, with clones FS22-172-003_AAA and FS22-172-003_APY completely losing binding and clones FS22-172-003_PPA and FS22-172-003_PAY retaining some binding, albeit significantly reduced (5.5- and 4.4-fold lower response units) compared to the parental clone. An impact on binding of FS22-053-008 was also observed, with the PAY and AAA variants completely losing binding and the APY and PPA variants showing reduced binding to the antigen and a slower association profile compared to the parental clone FS22-053-008. These data suggest that the PPY motif is important for the binding of both MAbs to the CD137 antigen.

13.3 Homology modeling

Given the involvement of the PPY motif in binding, in silico Fcab modeling was informative to evaluate protein structure predictions in the CH3 domain. Structural homology modeling and subsequent conformational searches were performed using the MOE software package, version 2019.0101 (Chemical Computing Group ULC). The Fc region of the 5JII structure from the Protein Data Bank [PDB] was used as a structural template for the Fcab region of both FS22-172-003 and FS22-053-008. Since there are insertions in the AB and EF loop regions relative to the chosen structural template, a de novo loop search with residue side chain optimization was used to obtain the AB and EF loop structures. The resulting homology models were energy minimized and scored according to geometric criteria including backbone bond lengths, angles, dihedral angles, and chirality. Structural homology models with scores meeting the criteria were used as the basis for conformational sampling using the LowModeMD modeling method implemented in MOE.

The homology models in Figure 13 show that the AB and EF loops of both Fcabs adopt different conformations compared to the 5JII template, with the PPY motif playing some role in the protrusion of the AB loops from the core CH3 domain. The PPY motif also mediates interactions that lead to conformational stabilization of the AB loop through both interactions within the loop and between the loop and the rest of the structure. Conformational searches performed using LowModeMD simulations also indicated that a tyrosine residue in the PPY motif interacts with Q3 (IMGT) and D12 (IMGT) in the CH3 domain of the same chain, in the CH3 domain, possibly stabilizing the AB loop. Taken together with the results shown in Example 13.2, this suggests that the PPY motif is important for the binding of both FS22-172-003 and FS22-053-008 to CD137.

List of sequences

Amino acid sequences of the structural loops of the CH3 domain of WT Fcab

Hinge AB WT Fcab - RDELTKNQ (SEQ ID NO: 1)

CD Loop WT Fcab - SNGQPENNY (SEQ ID NO: 2)

Hinge EF WT Fcab - DKSRWQQGNV (SEQ ID NO: 3)

Amino acid sequence of the CH3 domain of WT Fcab (SEQ ID NO: 4)

Loops AB, CD and EF are emphasized

GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the CH2 domain of Fcab with the LALA-PA mutation (SEQ ID NO: 5)

The LALA-PA mutation is underlined.

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAK

Amino acid sequence of the CH2 domain of Fcab with the LALA mutation (SEQ ID NO: 6)

The LALA mutation is underlined.

APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK

Amino acid sequence of the truncated hinge region of Fcab (SEQ ID NO: 7)

TCPPCP

Amino acid sequence of WT Fcab with LALA mutation (SEQ ID NO: 8)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

Amino acid sequence of WT Fcab without LALA mutation (SEQ ID NO: 9)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the PPY motif (SEQ ID NO: 10)

PPY

Amino acid sequence of Fcab FS22-033 with LALA mutation (SEQ ID NO: 11)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEYFEQEVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVAR HRWQLGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of Fcab FS22-033 without LALA mutation (SEQ ID NO: 12)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEYFEQEVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVAR HRWQLGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-033/HelD1.3 with the LALA mutation (SEQ ID NO: 13)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEYFEQEVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVAR HRWQLGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-033/HelD1.3 without the LALA mutation (SEQ ID NO: 14)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEYFEQEVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVAR HRWQLGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of Fcab FS22-053 with LALA mutation (SEQ ID NO: 15)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYYNRWQDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of Fcab FS22-053 without LALA mutation (SEQ ID NO: 16)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYYNRWQDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053/HelD1.3 with the LALA mutation (SEQ ID NO: 17)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYYNRWQDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053/HelD1.3 without the LALA mutation (SEQ ID NO: 18)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYYNRWQDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-053-008

First sequence FS22-053-008 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-008 – DYWRWLE (SEQ ID NO: 20)

Amino acid sequence of the CH3 domain of Fcab FS22-053-008 (SEQ ID NO: 21)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-008 (SEQ ID NO: 22)

GGACAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTACTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGATTACTGGAGG TGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGT

Amino acid sequence of Fcab FS22-053-008 with LALA mutation (SEQ ID NO: 23)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-008 with LALA mutation (SEQ ID NO: 24)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA TTACTGGAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-008 without LALA mutation (SEQ ID NO: 25)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-008 without LALA mutation (SEQ ID NO: 26)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA TTACTGGAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-008/HelD1.3 with the LALA mutation (SEQ ID NO: 27)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-008/HelD1.3 without the LALA mutation (SEQ ID NO: 28)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-053-009

First sequence FS22-053-009 - NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-009 - EHTRWLD (SEQ ID NO: 29)

Amino acid sequence of the CH3 domain of Fcab FS22-053-009 (SEQ ID NO: 30)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-009 (SEQ ID NO: 31)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGAACATACTAGG TGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-009 with LALA mutation (SEQ ID NO: 32)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-009 with LALA mutation (SEQ ID NO: 33)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA ACATACTAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-009 without LALA mutation (SEQ ID NO: 34)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-009 without LALA mutation (SEQ ID NO: 35)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA ACATACTAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-009/HelD1.3 with the LALA mutation (SEQ ID NO: 36)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-009/HelD1.3 without the LALA mutation (SEQ ID NO: 37)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-010

First sequence FS22-053-010 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-010 – DYMRWLD (SEQ ID NO: 38)

Amino acid sequence of the CH3 domain of Fcab FS22-053-010 (SEQ ID NO: 39)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDYMRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-010 (SEQ ID NO: 40)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGATTACATGAGG TGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-010 with LALA mutation (SEQ ID NO: 41)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYMRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-010 with LALA mutation (SEQ ID NO: 42)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATTACATGAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-010 without LALA mutation (SEQ ID NO: 43)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYMRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-010 without LALA mutation (SEQ ID NO: 44)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATTACATGAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-010/HelD1.3 with the LALA mutation (SEQ ID NO: 45)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYMRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-010/HelD1.3 without the LALA mutation (SEQ ID NO: 46)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYMRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-053-011

First sequence FS22-053-011 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-011 – DYWRWTD (SEQ ID NO: 47)

Amino acid sequence of the CH3 domain of Fcab FS22-053-011 (SEQ ID NO: 48)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-011 (SEQ ID NO: 49)

GGACAGCCTCGAGAGCCTCAAGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGATTACTGGAGG TGGACTGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGA

Amino acid sequence of Fcab FS22-053-011 with LALA mutation (SEQ ID NO: 50)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-011 with LALA mutation (SEQ ID NO: 51)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA TTACTGGAGGTGGACTGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-011 without LALA mutation (SEQ ID NO: 52)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-011 without LALA mutation (SEQ ID NO: 53)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGA TTACTGGAGGTGGACTGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-011/HelD1.3 with the LALA mutation (SEQ ID NO: 54)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-011/HelD1.3 without the LALA mutation (SEQ ID NO: 55)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-053-012

First sequence FS22-053-012 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-012 – DHMRWLE (SEQ ID NO: 56)

Amino acid sequence of the CH3 domain of Fcab FS22-053-012 (SEQ ID NO: 57)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDHMRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-012 (SEQ ID NO: 58)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGATCATATGAGG TGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-012 with LALA mutation (SEQ ID NO: 59)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDHMRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-012 with LALA mutation (SEQ ID NO: 60)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATCATATGAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-012 without LALA mutation (SEQ ID NO: 61)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDHMRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-012 without LALA mutation (SEQ ID NO: 62)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATCATATGAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-012/HelD1.3 with the LALA mutation (SEQ ID NO: 63)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDHMRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-012/HelD1.3 without the LALA mutation (SEQ ID NO: 64)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDHMRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-013

First sequence FS22-053-013 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-013 – GYERWLE (SEQ ID NO: 65)

Amino acid sequence of the CH3 domain of Fcab FS22-053-013 (SEQ ID NO: 66)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGYERWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-013 (SEQ ID NO: 67)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGTTACGAAAGG TGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-013 with LALA mutation (SEQ ID NO: 68)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGYERWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-013 with LALA mutation (SEQ ID NO: 69)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGG TTACGAAAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-013 without LALA mutation (SEQ ID NO: 70)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGYERWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-013 without LALA mutation (SEQ ID NO: 71)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGG TTACGAAAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-013/HelD1.3 with the LALA mutation (SEQ ID NO: 72)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGYERWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-013/HelD1.3 without the LALA mutation (SEQ ID NO: 73)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGYERWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-014

First sequence FS22-053-014 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-014 – YHWRWLD (SEQ ID NO: 74)

Amino acid sequence of the CH3 domain of Fcab FS22-053-014 (SEQ ID NO: 75)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVYHWRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-014 (SEQ ID NO: 76)

GGACAGCCTCGAGAGCCTCAAGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTACCATTGGAGG TGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGA

Amino acid sequence of Fcab FS22-053-014 with LALA mutation (SEQ ID NO: 77)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYHWRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-014 with LALA mutation (SEQ ID NO: 78)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTA CCATTGGAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-014 without LALA mutation (SEQ ID NO: 79)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYHWRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-014 without LALA mutation (SEQ ID NO: 80)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTA CCATTGGAGGTGGCTGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-014/HelD1.3 with the LALA mutation (SEQ ID NO: 81)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYHWRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-014/HelD1.3 without the LALA mutation (SEQ ID NO: 82)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYHWRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-015

First sequence FS22-053-015 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-015 – DHWRWLQ (SEQ ID NO: 83)

Amino acid sequence of the CH3 domain of Fcab FS22-053-015 (SEQ ID NO: 84)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDHWRWLQGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-015 (SEQ ID NO: 85)

Amino acid sequence of Fcab FS22-053-015 with LALA mutation (SEQ ID NO: 86)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDHWRWLQGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-015 with LALA mutation (SEQ ID NO: 87)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGTTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATCATTGGAGGTGGCTGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-015 without LALA mutation (SEQ ID NO: 88)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDHWRWLQGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-015 without LALA mutation (SEQ ID NO: 89)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGTTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATCATTGGAGGTGGCTGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-015/HelD1.3 with the LALA mutation (SEQ ID NO: 90)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDHWRWLQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-015/HelD1.3 without the LALA mutation (SEQ ID NO: 91)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDHWRWLQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-016

First sequence FS22-053-016 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-016 – DYIRWLN (SEQ ID NO: 92)

Amino acid sequence of the CH3 domain of Fcab FS22-053-016 (SEQ ID NO: 93)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDYIRWLNGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-016 (SEQ ID NO: 94)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGATTACATCAGG TGGCTGAACGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-016 with LALA mutation (SEQ ID NO: 95)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYIRWLNGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-016 with LALA mutation (SEQ ID NO: 96)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATTACATCAGGTGGCTGAACGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-053-016 without LALA mutation (SEQ ID NO: 97)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVDYIRWLNGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-016 without LALA mutation (SEQ ID NO: 98)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGG ATTACATCAGGTGGCTGAACGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-016/HelD1.3 with the LALA mutation (SEQ ID NO: 99)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYIRWLNGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-016/HelD1.3 without the LALA mutation (SEQ ID NO: 100)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYIRWLNGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-053-017

First sequence FS22-053-017 – NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053-017 – YHWRWLE (SEQ ID NO: 101)

Amino acid sequence of the CH3 domain of Fcab FS22-053-017 (SEQ ID NO: 102)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVYHWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053-017 (SEQ ID NO: 103)

GGACAGCCTCGAGAACCACAGGTGTACACTCTGCCCCCTTCACGCGACGAACTCAATCCGCCCTACCTGTTCTCCAACCAAGTCTCCCTGACCTGTCTTGTGAAGGGTTTCTACCCATCCGATATCGCCGTGGAGTGGGAGAGCAACGGACAGCCGGAGAACAACTATAAGACTACCCCGCCTGTGCTGGACTCGGACGGCAGCTTCTTCTTGTACTCCAAACTGACCGTGTACCACTGGCGG TGGCTGGAAGGGAACGTGTTTAGCTGCTCCGTCATGCATGAAGCCCTGCACAACCACTACACCCAGAAGTCCCTCTCGCTCTCTCCGGGT

Amino acid sequence of Fcab FS22-053-017 with LALA mutation (SEQ ID NO: 104)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYHWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-017 with LALA mutation (SEQ ID NO: 105)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGACAGCCTCGAGAACCACAGGTGTACACTCTGCCCCCTTCACGCGACGAACTCAATCCGCCCTACCTGTTCTCCAACCAAGTCTCCCTGACCTGTCTTGTGAAGGGTTTCTACCCATCCGATATCGCCGTGGAGTGGGAGAGCAACGGACAGCCGGAGAACAACTATAAGACTACCCCGCCTGTGCTGGACTCGGACGGCAGCTTCTTCTTGTACTCCAAACTGACCGTGTACC ACTGGCGGTGGCTGGAAGGGAACGTGTTTAGCTGCTCCGTCATGCATGAAGCCCTGCACAACCACTACACCCCAGAAGTCCCTCTCGCTCTCTCCGGGT

Amino acid sequence of Fcab FS22-053-017 without LALA mutation (SEQ ID NO: 106)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVYHWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-053-017 without LALA mutation (SEQ ID NO: 107)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGACAGCCTCGAGAACCACAGGTGTACACTCTGCCCCCTTCACGCGACGAACTCAATCCGCCCTACCTGTTCTCCAACCAAGTCTCCCTGACCTGTCTTGTGAAGGGTTTCTACCCATCCGATATCGCCGTGGAGTGGGAGAGCAACGGACAGCCGGAGAACAACTATAAGACTACCCCGCCTGTGCTGGACTCGGACGGCAGCTTCTTCTTGTACTCCAAACTGACCGTGTACC ACTGGCGGTGGCTGGAAGGGAACGTGTTTAGCTGCTCCGTCATGCATGAAGCCCTGCACAACCACTACACCCCAGAAGTCCCTCTCGCTCTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-017/HelD1.3 with the LALA mutation (SEQ ID NO: 108)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYHWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-053-017/HelD1.3 without the LALA mutation (SEQ ID NO: 109)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVYHWRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172

First sequence FS22-172 – RKYYPPY (SEQ ID NO: 110)

Second sequence FS22-172 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172 (SEQ ID NO: 112)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELRKYYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172 (SEQ ID NO: 113)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172 with LALA mutation (SEQ ID NO: 114)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172 with LALA mutation (SEQ ID NO: 115)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172 without LALA mutation (SEQ ID NO: 116)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172 without LALA mutation (SEQ ID NO: 117)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172/HelD1.3 with the LALA mutation (SEQ ID NO: 118)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172/HelD1.3 without the LALA mutation (SEQ ID NO: 119)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loop sequences of the CH3 domain of Fcab FS22-172-001

First sequence FS22-172-001 – PFVMPPY (SEQ ID NO: 120)

Second sequence FS22-172-001 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-001 (SEQ ID NO: 121)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELPFVMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-001 (SEQ ID NO: 122)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCGTTATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-001 with LALA mutation (SEQ ID NO: 123)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFVMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-001 with LALA mutation (SEQ ID NO: 124)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCGTTATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-001 without LALA mutation (SEQ ID NO: 125)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFVMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-001 without LALA mutation (SEQ ID NO: 126)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCGTTATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-001/HelD1.3 with the LALA mutation (SEQ ID NO: 127)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFVMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-001/HelD1.3 without the LALA mutation (SEQ ID NO: 128)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFVMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172-002

First sequence FS22-172-002 – PFQMPPY (SEQ ID NO: 129)

Second sequence FS22-172-002 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-002 (SEQ ID NO: 130)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-002 (SEQ ID NO: 131)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCCAGATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-002 with LALA mutation (SEQ ID NO: 132)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-002 with LALA mutation (SEQ ID NO: 133)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCCAGATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-002 without LALA mutation (SEQ ID NO: 134)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-002 without LALA mutation (SEQ ID NO: 135)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATTCCAGATGCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-002/HelD1.3 with the LALA mutation (SEQ ID NO: 136)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-002/HelD1.3 without the LALA mutation (SEQ ID NO: 137)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172-003

The first sequence FS22-172-003 is PYIIPPY (SEQ ID NO: 138)

Second sequence FS22-172-003 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-003 (SEQ ID NO: 139)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-003 (SEQ ID NO: 140)

GGACAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATACATCATCCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTGGCTG GAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGT

Amino acid sequence of Fcab FS22-172-003 with LALA mutation (SEQ ID NO: 141)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-003 with LALA mutation (SEQ ID NO: 142)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGACAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATACATCATCCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAG ATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGT

Amino acid sequence of Fcab FS22-172-003 without LALA mutation (SEQ ID NO: 143)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-003 without LALA mutation (SEQ ID NO: 144)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGACAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCCATACATCATCCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAG ATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-003/HelD1.3 with the LALA mutation (SEQ ID NO: 145)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-003/HelD1.3 without the LALA mutation (SEQ ID NO: 146)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172-004

First sequence FS22-172-004 – NYIYPPY (SEQ ID NO: 147)

Second sequence FS22-172-004 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-004 (SEQ ID NO: 148)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-004 (SEQ ID NO: 149)

GGACAGCCTCGAGAGCCTCAAGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACTACATCTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCGCTGCACAACCACTACACTCAGAAGAGCTTGTCCCTGTCGCCCGGA

Amino acid sequence of Fcab FS22-172-004 with LALA mutation (SEQ ID NO: 150)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-004 with LALA mutation (SEQ ID NO: 151)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACTACATCTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-004 without LALA mutation (SEQ ID NO: 152)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-004 without LALA mutation (SEQ ID NO: 153)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACTACATCTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-004/HelD1.3 with the LALA mutation (SEQ ID NO: 154)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-004/HelD1.3 without the LALA mutation (SEQ ID NO: 155)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172-005

First sequence FS22-172-005 – QQVYPPY (SEQ ID NO: 156)

Second sequence FS22-172-005 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-005 (SEQ ID NO: 157)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELQQVYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-005 (SEQ ID NO: 158)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCAGCAGGTTTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-005 with LALA mutation (SEQ ID NO: 159)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELQQVYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-005 with LALA mutation (SEQ ID NO: 160)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCAGCAGGTTTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-005 without LALA mutation (SEQ ID NO: 161)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELQQVYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-005 without LALA mutation (SEQ ID NO: 162)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCAGCAGGTTTACCCACCATACAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-005/HelD1.3 with the LALA mutation (SEQ ID NO: 163)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELQQVYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-005/HelD1.3 without the LALA mutation (SEQ ID NO: 164)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELQQVYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-172-006

First sequence FS22-172-006 – RKYYPPY (SEQ ID NO: 110)

Second sequence FS22-172-006 – GADRWLE (SEQ ID NO: 111)

Amino acid sequence of the CH3 domain of Fcab FS22-172-006 (SEQ ID NO: 165)

The first and second sequences are underlined.

GQPREPQVYTLPPSRDELRKYYPPYNQLSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-172-006 (SEQ ID NO: 166)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGCTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGCGCAGATAGGTG GCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-006 with LALA mutation (SEQ ID NO: 167)

Hinge region (underlined), CH2 domain (bold), CH3 domain (italic), LALA mutation (bold and underlined)

TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQLSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-006 with LALA mutation (SEQ ID NO: 168)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGCTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of Fcab FS22-172-006 without LALA mutation (SEQ ID NO: 169)

Hinge region (underlined), CH2 domain (bold), and CH3 domain (italic)

TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQLSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of Fcab FS22-172-006 without LALA mutation (SEQ ID NO: 170)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGCGTAAATACTACCCGCCGTACAACCAGCTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGGC GCAGATAGGTGGCTGGAAGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-006/HelD1.3 with the LALA mutation (SEQ ID NO: 171)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQLSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain of the mimic MAb ² FS22-172-006/HelD1.3 without the LALA mutation (SEQ ID NO: 172)

VH domain (underlined)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELRKYYPPYNQLSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the light chain of the HelD1.3 mimic MAb ² (SEQ ID NO: 173)

VL domain (underlined)

DIQMTQSPASLSSASVGETVTITCRASGNIHNYLAWYQQKQGKSPQLLVYNAKTLADGVPSRFSGSGSGTQYSLKINSLQPEDFGSYYCQHFWSTPRTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC

Amino acid sequences of the structural loops of the CH3 domain of Fcab FS22-053

First sequence FS22-053 - NPPYLFS (SEQ ID NO: 19)

Second sequence FS22-053 - YYNRWQD (SEQ ID NO: 174)

Amino acid sequence of the CH3 domain of Fcab FS22-053 (SEQ ID NO: 175) (first and second sequences are underlined).

GQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVYYNRWQDGNVFSCSVMHEALHNHYTQKSLSLSPG

Nucleic acid sequence of the CH3 domain of Fcab FS22-053 (SEQ ID NO: 176)

GGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTATTATAACAGG TGGCAGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCCGGGT

Nucleic acid sequence of Fcab FS22-053 Fcab with LALA (SEQ ID NO: 177)

ACTTGCCCGCCTTGCCCAGCCCCGGAAGCTGCCGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGG TGTCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTA TTATAACAGGTGGCAGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Nucleic acid sequence of Fcab FS22-053 Fcab without LALA (SEQ ID NO: 178)

ACTTGCCCGCCTTGCCCAGCCCCGGAACTGCTGGGTGGTCCTTCGGTGTTCCTCTTCCCGCCCAAGCCGAAGGATACCCTGATGATCTCACGGACCCCCGAAGTGACCTGTGTGGTGGTGGACGTGTCCCACGAGGACCCGGAAGTGAAATTCAATTGGTACGTGGATGGAGTGGAAGTGCACAACGCCAAGACCAAGCCACGGGAAGAACAGTACAACTCTACCTACCGCGTGGTG TCCGTGCTCACTGTGCTGCACCAAGACTGGCTGAACGGGAAGGAGTACAAGTGCAAAGTGTCCAACAAGGCGCTGCCTGCCCCAATTGAGAAAACTATCTCGA AAGCCAAGGGCCAGCCTCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGAACCCGCCGTACCTGTTCTCTAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGTA TTATAACAGGTGGCAGGATGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAGAGCCTCTCCCTGTCTCCGGGT

Amino acid sequences of the full-length immunoglobulin hinge region (SEQ ID NO: 179)

EPKSCDKTHTCPPCP

Amino acid sequence of human CD137 (SEQ ID NO: 180)

Extracellular domain (italic); transmembrane and intracellular domains (bold)

lqdpcsncpagtfcdnnrnqicspcppnsfssaggqrtcdicrqckgvfrtrkecsstsnaecdctpgfhclgagcsmceqdckqgqeltkkgckdccfgtfndqkrgicrpwtncsldgksvlvngtkerdvvcgpspadlspgassvtppaparepghspqIISFFLALTSTALLFLLFFL TLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL

Amino acid sequence of the extracellular domain of human CD137 (SEQ ID NO: 181)

LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQ

Amino acid sequence of CD137 from cynomolgus monkey (SEQ ID NO: 182)

Extracellular domain (italic); transmembrane and intracellular domains (bold)

LQDLCSNCPAGTFCDNNRSQICSPCPPNSFSSAGGQRTCDICRQCKGVFKTRKECSSTSNAECDCISGYHCLGAECSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSATPPAPAREPGHSPQIIFFLALTSTVVLFLLFFLVLRFSVVKRSRKKLLYIFKQ PFMRPVQTTQEEDGCSCRFPEEEEGGCEL

Amino acid sequence of the extracellular domain of CD137 from cynomolgus monkey (SEQ ID NO: 183)

LQDLCSNCPAGTFCDNNRSQICSPCPPNSFSSAGGQRTCDICRQCKGVFKTRKECSSTSNAECDCISGYHCLGAECSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSATPPAPAREPGHSPQ

Amino acid sequence of mouse CD137 (SEQ ID NO: 184)

Extracellular domain (italic); transmembrane and intracellular domains (bold)

vqnscdncqpgtfcrkynpvckscppstfssiggqpncnicrvcagyfrfkkfcssthnaececiegfhclgpqctrcekdcrpgqeltkqgcktcslgtfndqngtgvcrpwtncsldgrsvlktgttekdvvcgppvvsfspsttisvtpeggpgghslqvlTLFLALTS ALLLALIFITLLFSVLKWIRKKFPHIFKQPFKKTTGAAQEEDACSCRCPQEEEGGGGGYEL

Amino acid sequence of the extracellular domain of mouse CD137 (SEQ ID NO: 185)

VQNSCDNCQPGTFCRKYNPVCKSCPPSTFSSIGGQPNCNICRVCAGYFRFKKFCSSTHNAECECIEGFHCLGPQCTRCEKDCRPGQELTKQGCKTCSLGTFNDQNGTGVCRPWTNCSLDGRSVLKTGTTEKDVVCGPPVVSFSPSTTISVTPEGGPGGHSLQVL

Amino acid sequence of the heavy chain of MAT G1/HelD1.3 (SEQ ID NO: 186)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of human PD-L1 (SEQ ID NO: 187)

MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEK LFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET

Amino acid sequence of mouse PD-L1 (SEQ ID NO: 188)

MRIFAGIIFTACCHLLRAFTITAPKDLYVVEYGSNVTMECRFPVERELDLLALVVYWEKEDEQVIQFVAGEEDLKPQHSNFRGRASLPKDQLLKGNAALQITDVKLQDAGVYCCIISYGGADYKRITLKVNAPYRKINQRISVDPATSEHELICQAEGYPEAEVIWTNSDHQPVSGKRSVTTSRTEGMLLNVTSSLR VNATANDVFYCTFWRSQPGQNHTAELIIPELPATHPPQNRTHWVLLGSILLFLIVVSTVLLFLRKQVRMLDVEKCGVEDTSSKNRNDTQFEET

Amino acid sequence of mouse mesothelin (SEQ ID NO: 189)

DAEQKACPPGKEPYKVDEDLIFYQNWELEACVDGTMLARQMDLVNEIPFTYEQLSIFKHKLDKTYPQGYPESLIQQLGHFFRYVSPEDIHQWNVTSPDTVKTLLKVSKGQKMNAQAIALVACYLRGGGQLDEDMVKALGDIPLSYLCDFSPQDLHSVPSSVMWLVGPQDLDKCSQRHLGLLYQKA CSAFQNVSGLEYFEKIKTFLGGASVKDLRALSQHNVSMDIATFKRLQVDSLVGLSVAEVQKLLGPNIVDLKTEEDKSPVRDWLFRQHQKDLDRLGLGLQGGIPNGYLVLDFNVREAFS

Amino acid sequence of the heavy chain of MAT ² FS22m-063-AA/FS28m-228 (SEQ ID NO: 190)

EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYFMVWVRQAPGKGLEWVSMISPKSSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARWFTPARFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEPYWSYVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VMNYRWELGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the light chain of MAT ² FS22m-063-AA/FS28m-228 (SEQ ID NO: 191)

EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQPFPFSFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE VTHQGLSSPVTKSFNRGEC

Amino acid sequence of the heavy chain of MAT G1AA/HelD1.3 (with LALA) (SEQ ID NO: 192)

QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVRQPPGRGLEWIGMIWGDGNTDYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain FS22-172-004-AA/S70 (with LALA) (SEQ ID NO: 193)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy chain amino acid sequence of FS22-172-004-AA/S70 (without LALA) (SEQ ID NO: 194)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNYIYPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Light chain amino acid sequence FS22-172-004-AA/S70 (SEQ ID NO: 194)

DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC

Amino acid sequence of the heavy chain FS22-172-003-AA/S70 (with LALA) (SEQ ID NO: 195)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy chain amino acid sequence of FS22-172-003-AA/S70 (without LALA) (SEQ ID NO: 196)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPYIIPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Amino acid sequence of the heavy chain FS22-172-002-AA/S70 (with LALA) (SEQ ID NO: 197)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy chain amino acid sequence FS22-172-002-AA/S70 (without LALA) (SEQ ID NO: 198)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-172-002-AA/S70 (without LALA) (SEQ ID NO: 199)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-172-002-AA/S70 (with LALA) (SEQ ID NO 200)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELPFQMPPYNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLY SKLTVGADRWLEGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-053-011-AA/S70 (without LALA) (SEQ ID NO: 201)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-053-011-AA/S70 (with LALA) (SEQ ID NO: 202)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVDYWRWTDGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-053-008-AA/S70 (with LALA) (SEQ ID NO: 203)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22-053-008-AA/S70 (without LALA) (SEQ ID NO: 204)

EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHT CPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELNPPYLFSNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS KLTVEHTRWLDGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22m-063-AA/F2 (without LALA) (SEQ ID NO: 205)

EVQLVQSGGGVVQPGRSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKEENWGSYFDLWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEPYWSYVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VMNYRWELGNVFSCSVMHEALHNHYTQKSLSLSPG

Heavy Chain FS22m-063-AA/F2 (without LALA) (SEQ ID NO: 206)

EVQLVQSGGGVVQPGRSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKEENWGSYFDLWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSCDK THTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEPYWSYVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VMNYRWELGNVFSCSVMHEALHNHYTQKSLSLSPG

Light chain FS22m-063-AA/F2 (SEQ ID NO: 207)

DIVMTQSPSTLSASVGDRVTITCRASQGISSWLAWYQQKPGRAPKVLIYKASTLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC

Heavy Chain FS22m-063/FS28m-228 (without LALA) (SEQ ID NO: 208)

EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYFMVWVRQAPGKGLEWVSMISPKSSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARWFTPARFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDEPYWSYVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VMNYRWELGNVFSCSVMHEALHNHYTQKSLSLSPG

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Wen T, Bukcyznski J, Watts TH., 2002. 4-1BB Ligand-Mediated Costimulation of Human T Cells Induces CD4 and CD8 T Cell Expansion, Cytokine Production, and the Development of Cytolytic Effector Function. The Journal of Immunology, 168(10):4897–906.

Wesche-Soldato DE, Chung CS, Gregory SH, Salazar-Mather TP, Ayala CA, Ayala A. CD8+ T cells promote inflammation and apoptosis in the liver after sepsis: role of Fas-FasL. Am. J. Pathol. 171(1), 87-96 (2007).

Westwood, J.A. et al., 2014. Combination anti-CD137 and anti-CD40 antibody therapy in murine myc-driven hematological cancers. Leukemia Research, 38(8), pp.948–954.

Won, E.-Y. et al., 2009. The structure of the trimer of human 4-1BB ligand is unique among members of the tumor necrosis factor superfamily. The Journal of biological chemistry, 285(12), pp. 9202–9210.

Wozniak-Knopp G, Bartl S, Bauer A, Mostageer M, Woisetschläger M, Antes B, Ettl K, Kainer M, Weberhofer G, Wiederkum S, Himmler G, Mudde GC, Rüker F. Introducing antigen-binding sites in structural loops of immunoglobulin constant domains: Fc fragments with engineered HER2/neu-binding sites and antibody properties. Protein Eng. Des. Sel. 23(4), 289-97 (2010).

Ye Q, Song D, Poussin M, Yamamoto T, Best A, Li C, Coukos G, Powell Jr D. 2014. CD137 accurately identifies and enriches for naturally occurring tumor-reactive T cells in tumor. Clin Cancer Res 20(1):44–55.

--->

List of sequences

<110> F-STAR BETA LIMITED

<120> FC-binding fragments containing an antigen-binding site

for CD137

<130> 007508138

<140> PCT/EP2019/068803

<141> 2019-07-12

<150>GB 1811408.2

<151> 2018-07-12

<160> 240

<170> PatentIn version 3.5

<210> 1

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<223> Loop AB WT Fcab

<400> 1

Arg Asp Glu Leu Thr Lys Asn Gln

1 5

<210> 2

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> CD WT Fcab Loop

<400> 2

Ser Asn Gly Gln Pro Glu Asn Asn Tyr

1 5

<210> 3

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<223> EF WT Fcab Loop

<400> 3

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

1 5 10

<210> 4

<211> 106

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 WT Fcab Domain

<400> 4

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

20 25 30

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

35 40 45

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

50 55 60

Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly

65 70 75 80

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

85 90 95

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105

<210> 5

<211> 110

<212> Protein

<213> Artificial sequence

<220>

<223> CH2 Fcab domain with LALA-PA mutation

<400> 5

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

Ala Leu Ala Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

100 105 110

<210> 6

<211> 110

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab CH2 domain with LALA mutation

<400> 6

Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

1 5 10 15

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

20 25 30

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

35 40 45

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

50 55 60

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

65 70 75 80

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

85 90 95

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys

100 105 110

<210> 7

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<223> Truncated hinge region Fcab

<400> 7

Thr Cys Pro Pro Cys Pro

1 5

<210> 8

<211> 223

<212> Protein

<213> Artificial sequence

<220>

<223> WT Fcab with LALA mutation

<400> 8

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

130 135 140

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

145 150 155 160

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

165 170 175

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

180 185 190

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

195 200 205

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

210 215 220

<210> 9

<211> 222

<212> Protein

<213> Artificial sequence

<220>

<223> WT Fcab without LALA mutation

<400> 9

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

130 135 140

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

145 150 155 160

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

165 170 175

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg

180 185 190

Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

195 200 205

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

210 215 220

<210> 10

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> PPY Motive

<400> 10

Pro Pro Tyr

1

<210> 11

<211> 222

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-033 with LALA mutation

<400> 11

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Tyr Phe Glu Gln Glu Val Ser Leu Thr Cys Leu

130 135 140

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

145 150 155 160

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

165 170 175

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ala Arg His Arg

180 185 190

Trp Gln Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

195 200 205

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

210 215 220

<210> 12

<211> 222

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-033 without LALA mutation

<400> 12

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Tyr Phe Glu Gln Glu Val Ser Leu Thr Cys Leu

130 135 140

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

145 150 155 160

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

165 170 175

Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ala Arg His Arg

180 185 190

Trp Gln Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

195 200 205

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

210 215 220

<210> 13

<211> 445

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-033/HelD1.3 with LALA mutation

<400> 13

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Tyr Phe Glu Gln Glu Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ala Arg His Arg Trp

405 410 415

Gln Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 14

<211> 445

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-033/HelD1.3 without LALA mutation

<400> 14

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Tyr Phe Glu Gln Glu Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Ala Arg His Arg Trp

405 410 415

Gln Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 15

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053 with LALA mutation

<400> 15

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr Tyr Asn Arg Trp Gln Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 16

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053 without LALA mutation

<400> 16

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr Tyr Asn Arg Trp Gln Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 17

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053/HelD1.3 with LALA mutation

<400> 17

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr Tyr Asn Arg Trp Gln Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 18

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053/HelD1.3 without LALA mutation

<400> 18

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr Tyr Asn Arg Trp Gln Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 19

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-008

<400> 19

Asn Pro Pro Tyr Leu Phe Ser

1 5

<210> 20

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-008

<400> 20

Asp Tyr Trp Arg Trp Leu Glu

1 5

<210> 21

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-008

<400> 21

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Tyr Trp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 22

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-008

<400> 22

ggacagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtactgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggattactgg 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggcgct gcacaaccac 300

tacactcaga agagcttgtc cctgtcgccc ggt 333

<210> 23

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-008 with LALA mutation

<400> 23

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 24

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-008 with LALA mutation

<400> 24

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attactggag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 25

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-008 without LALA mutation

<400> 25

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 26

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-008 without LALA mutation

<400> 26

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attactggag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 27

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-008/HelD1.3 with LALA mutation

<400> 27

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 28

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-008/HelD1.3 without LALA mutation

<400> 28

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 29

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-009

<400> 29

Glu His Thr Arg Trp Leu Asp

1 5

<210> 30

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-009

<400> 30

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Glu His Thr

65 70 75 80

Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 31

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-009

<400> 31

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggaacatact 240

aggtggctgg atgggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 32

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-009 with LALA mutation

<400> 32

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 33

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-009 with LALA mutation

<400> 33

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg aacatactag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 34

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-009 without LALA mutation

<400> 34

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 35

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-009 without LALA mutation

<400> 35

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg aacatactag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 36

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-009/HelD1.3 with LALA mutation

<400> 36

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 37

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-009/HelD1.3 without LALA mutation

<400> 37

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 38

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence Fcab FS22-053-010

<400> 38

Asp Tyr Met Arg Trp Leu Asp

1 5

<210> 39

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-010

<400> 39

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Tyr Met

65 70 75 80

Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 40

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-010

<400> 40

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggattacatg 240

aggtggctgg atgggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 41

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-010 with LALA mutation

<400> 41

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Met Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 42

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-010 with LALA mutation

<400> 42

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attacatgag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 43

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-010 without LALA mutation

<400> 43

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Met Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 44

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-010 without LALA mutation

<400> 44

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attacatgag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 45

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-010/HelD1.3 with LALA mutation

<400> 45

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Met Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 46

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-010/HelD1.3 without LALA mutation

<400> 46

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Met Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 47

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-011

<400> 47

Asp Tyr Trp Arg Trp Thr Asp

1 5

<210> 48

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-011

<400> 48

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Tyr Trp

65 70 75 80

Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 49

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-011

<400> 49

ggacagcctc gagagcctca agtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggattactgg 240

aggtggactg atgggaacgt cttctcatgc tccgtgatgc atgaggcgct gcacaaccac 300

tacactcaga agagcttgtc cctgtcgccc gga 333

<210> 50

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-011 with LALA mutation

<400> 50

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 51

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-011 with LALA mutation

<400> 51

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attactggag gtggactgat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 52

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-011 without LALA mutation

<400> 52

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 53

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-011 without LALA mutation

<400> 53

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attactggag gtggactgat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 54

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-011/HelD1.3 with LALA mutation

<400> 54

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 55

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-011/HelD1.3 without LALA mutation

<400> 55

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 56

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-012

<400> 56

Asp His Met Arg Trp Leu Glu

1 5

<210> 57

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-012

<400> 57

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp His Met

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 58

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-012

<400> 58

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggatcatatg 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 59

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-012 with LALA mutation

<400> 59

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp His Met Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 60

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-012 with LALA mutation

<400> 60

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg atcatatgag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 61

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-012 without LALA mutation

<400> 61

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp His Met Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 62

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-012 without LALA mutation

<400> 62

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg atcatatgag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 63

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-012/HelD1.3 with LALA mutation

<400> 63

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp His Met Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 64

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-012/HelD1.3 without LALA mutation

<400> 64

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp His Met Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 65

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-013

<400> 65

Gly Tyr Glu Arg Trp Leu Glu

1 5

<210> 66

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-013

<400> 66

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Tyr Glu

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 67

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-013

<400> 67

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggttacgaa 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 68

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-013 with LALA mutation

<400> 68

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Tyr Glu Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 69

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-013 with LALA mutation

<400> 69

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gttacgaaag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 70

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-013 without LALA mutation

<400> 70

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Tyr Glu Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 71

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-013 without LALA mutation

<400> 71

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gttacgaaag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 72

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-013/HelD1.3 with LALA mutation

<400> 72

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Tyr Glu Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 73

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-013/HelD1.3 without LALA mutation

<400> 73

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Tyr Glu Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 74

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-014

<400> 74

Tyr His Trp Arg Trp Leu Asp

1 5

<210> 75

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-014

<400> 75

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Tyr His Trp

65 70 75 80

Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 76

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-014

<400> 76

ggacagcctc gagagcctca agtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gtaccattgg 240

aggtggctgg atgggaacgt cttctcatgc tccgtgatgc atgaggcgct gcacaaccac 300

tacactcaga agagcttgtc cctgtcgccc gga 333

<210> 77

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-014 with LALA mutation

<400> 77

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr His Trp Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 78

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-014 with LALA mutation

<400> 78

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgt accattggag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 79

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-014 without LALA mutation

<400> 79

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr His Trp Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 80

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-014 without LALA mutation

<400> 80

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgt accattggag gtggctggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 81

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-014/HelD1.3 with LALA mutation

<400> 81

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr His Trp Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 82

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-014/HelD1.3 without LALA mutation

<400> 82

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr His Trp Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 83

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-015

<400> 83

Asp His Trp Arg Trp Leu Gln

1 5

<210> 84

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-015

<400> 84

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp His Trp

65 70 75 80

Arg Trp Leu Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 85

<400> 85

000

<210> 86

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-015 with LALA mutation

<400> 86

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp His Trp Arg Trp Leu Gln Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 87

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-015 with LALA mutation

<400> 87

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgttggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg atcattggag gtggctgcag 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 88

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-015 without LALA mutation

<400> 88

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp His Trp Arg Trp Leu Gln Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 89

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-015 without LALA mutation

<400> 89

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgttggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg atcattggag gtggctgcag 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 90

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-015/HelD1.3 with LALA mutation

<400> 90

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp His Trp Arg Trp Leu Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 91

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-015/HelD1.3 without LALA mutation

<400> 91

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp His Trp Arg Trp Leu Gln Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 92

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-016

<400> 92

Asp Tyr Ile Arg Trp Leu Asn

1 5

<210> 93

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-016

<400> 93

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Tyr Ile

65 70 75 80

Arg Trp Leu Asn Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 94

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-016

<400> 94

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggattacatc 240

aggtggctga acgggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 95

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-016 with LALA mutation

<400> 95

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Ile Arg Trp Leu Asn Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 96

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-016 with LALA mutation

<400> 96

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attacatcag gtggctgaac 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 97

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-016 without LALA mutation

<400> 97

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Asp Tyr Ile Arg Trp Leu Asn Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 98

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-016 without LALA mutation

<400> 98

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg attacatcag gtggctgaac 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 99

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-016/HelD1.3 with LALA mutation

<400> 99

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Ile Arg Trp Leu Asn Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 100

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-016/HelD1.3 without LALA mutation

<400> 100

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Asp Tyr Ile Arg Trp Leu Asn Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 101

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053-017

<400> 101

Tyr His Trp Arg Trp Leu Glu

1 5

<210> 102

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-017

<400> 102

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Tyr His Trp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 103

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053-017

<400> 103

ggacagcctc gagaaccaca ggtgtacact ctgccccctt cacgcgacga actcaatccg 60

ccctacctgt tctccaacca agtctccctg acctgtcttg tgaagggttt ctacccatcc 120

gatatcgccg tggagtggga gagcaacgga cagccggaga acaactataa gactaccccg 180

cctgtgctgg actcggacgg cagcttcttc ttgtactcca aactgaccgt gtaccactgg 240

cggtggctgg aagggaacgt gtttagctgc tccgtcatgc atgaagccct gcacaaccac 300

tacacccaga agtccctctc gctctctccg ggt 333

<210> 104

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-017 with LALA mutation

<400> 104

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr His Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 105

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-017 with LALA mutation

<400> 105

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg acagcctcga 360

gaaccacagg tgtacactct gcccccttca cgcgacgaac tcaatccgcc ctacctgttc 420

tccaaccaag tctccctgac ctgtcttgtg aagggtttct acccatccga tatcgccgtg 480

gagtgggaga gcaacggaca gccggagaac aactataaga ctaccccgcc tgtgctggac 540

tcggacggca gcttcttctt gtactccaaa ctgaccgtgt accactggcg gtggctggaa 600

gggaacgtgt ttagctgctc cgtcatgcat gaagccctgc acaaccacta cacccagaag 660

tccctctcgc tctctccggg t 681

<210> 106

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-053-017 without LALA mutation

<400> 106

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Tyr His Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 107

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053-017 without LALA mutation

<400> 107

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg acagcctcga 360

gaaccacagg tgtacactct gcccccttca cgcgacgaac tcaatccgcc ctacctgttc 420

tccaaccaag tctccctgac ctgtcttgtg aagggtttct acccatccga tatcgccgtg 480

gagtgggaga gcaacggaca gccggagaac aactataaga ctaccccgcc tgtgctggac 540

tcggacggca gcttcttctt gtactccaaa ctgaccgtgt accactggcg gtggctggaa 600

gggaacgtgt ttagctgctc cgtcatgcat gaagccctgc acaaccacta cacccagaag 660

tccctctcgc tctctccggg t 681

<210> 108

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-017/HelD1.3 with LALA mutation

<400> 108

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr His Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 109

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-053-017/HelD1.3 without LALA mutation

<400> 109

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Tyr His Trp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 110

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> Structural loop sequence of the CH3 domain of Fcab FS22-172

<400> 110

Arg Lys Tyr Tyr Pro Pro Tyr

1 5

<210> 111

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> Structural loop sequence of the CH3 domain of Fcab FS22-172

<400> 111

Gly Ala Asp Arg Trp Leu Glu

1 5

<210> 112

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172

<400> 112

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 113

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172

<400> 113

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgcgtaaa 60

tactacccgc cgtacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 114

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172 with LALA mutation

<400> 114

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 115

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172 with LALA mutation

<400> 115

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcgtaaata ctacccgccg 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 116

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172 without LALA mutation

<400> 116

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 117

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172 without LALA mutation

<400> 117

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcgtaaata ctacccgccg 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 118

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172/HelD1.3 with LALA mutation

<400> 118

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 119

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172/HelD1.3 without LALA mutation

<400> 119

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 120

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-172-001

<400> 120

Pro Phe Val Met Pro Pro Tyr

1 5

<210> 121

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-001

<400> 121

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Pro Phe Val Met Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 122

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-001

<400> 122

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgccattc 60

gttatgccac catacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 123

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-001 with LALA mutation

<400> 123

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Phe Val Met Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 124

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-001 with LALA mutation

<400> 124

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccattcgt tatgccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 125

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-001 without LALA mutation

<400> 125

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Phe Val Met Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 126

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-001 without LALA mutation

<400> 126

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccattcgt tatgccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 127

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-001/HelD1.3 with LALA mutation

<400> 127

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Phe Val Met Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 128

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-001/HelD1.3 without LALA mutation

<400> 128

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Phe Val Met Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 129

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-172-002

<400> 129

Pro Phe Gln Met Pro Pro Tyr

1 5

<210> 130

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-002

<400> 130

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 131

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-002

<400> 131

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgccattc 60

cagatgccac catacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 132

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-002 with LALA mutation

<400> 132

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 133

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-002 with LALA mutation

<400> 133

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccattcca gatgccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 134

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-002 without LALA mutation

<400> 134

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 135

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-002 without LALA mutation

<400> 135

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccattcca gatgccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 136

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-002/HelD1.3 with LALA mutation

<400> 136

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 137

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-002/HelD1.3 without LALA mutation

<400> 137

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 138

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-172-003

<400> 138

Pro Tyr Ile Ile Pro Pro Tyr

1 5

<210> 139

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-003

<400> 139

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 140

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-003

<400> 140

ggacagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgccatac 60

atcatcccac catacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggcgct gcacaaccac 300

tacactcaga agagcttgtc cctgtcgccc ggt 333

<210> 141

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-003 with LALA mutation

<400> 141

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 142

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-003 with LALA mutation

<400> 142

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg acagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccatacat catcccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggcgctgc acaaccacta cactcagaag 660

agcttgtccc tgtcgcccgg t 681

<210> 143

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-003 without LALA mutation

<400> 143

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 144

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-003 without LALA mutation

<400> 144

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg acagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgccatacat catcccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggcgctgc acaaccacta cactcagaag 660

agcttgtccc tgtcgcccgg t 681

<210> 145

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-003/HelD1.3 with LALA mutation

<400> 145

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 146

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-003/HelD1.3 without LALA mutation

<400> 146

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 147

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-172-004

<400> 147

Asn Tyr Ile Tyr Pro Pro Tyr

1 5

<210> 148

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-004

<400> 148

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 149

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-004

<400> 149

ggacagcctc gagagcctca agtgtacacc ctgcccccat cccgggatga gctgaactac 60

atctacccac catacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggcgct gcacaaccac 300

tacactcaga agagcttgtc cctgtcgccc gga 333

<210> 150

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-004 with LALA mutation

<400> 150

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 151

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-004 with LALA mutation

<400> 151

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaactacat ctacccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 152

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-004 without LALA mutation

<400> 152

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 153

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-004 without LALA mutation

<400> 153

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaactacat ctacccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 154

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-004/HelD1.3 with LALA mutation

<400> 154

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 155

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-004/HelD1.3 without LALA mutation

<400> 155

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 156

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-172-005

<400> 156

Gln Gln Val Tyr Pro Pro Tyr

1 5

<210> 157

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-005

<400> 157

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Gln Gln Val Tyr Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 158

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-005

<400> 158

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgcagcag 60

gtttacccac catacaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 159

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-005 with LALA mutation

<400> 159

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Gln Gln Val Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 160

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-005 with LALA mutation

<400> 160

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcagcaggt ttacccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 161

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-005 without LALA mutation

<400> 161

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Gln Gln Val Tyr Pro Pro Tyr Asn Gln Val

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 162

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-005 without LALA mutation

<400> 162

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcagcaggt ttacccacca 420

tacaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 163

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-005/HelD1.3 with LALA mutation

<400> 163

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Gln Gln Val Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 164

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-005/HelD1.3 without LALA mutation

<400> 164

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Gln Gln Val Tyr Pro Pro Tyr Asn Gln Val Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 165

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-006

<400> 165

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Leu Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp

65 70 75 80

Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 166

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-172-006

<400> 166

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgcgtaaa 60

tactacccgc cgtacaacca gctcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gggcgcagat 240

aggtggctgg aagggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 167

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-006 with LALA mutation

<400> 167

Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Leu

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 168

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-006 with LALA mutation

<400> 168

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcgtaaata ctacccgccg 420

tacaaccagc tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 169

<211> 227

<212> Protein

<213> Artificial sequence

<220>

<223> Fcab FS22-172-006 without LALA mutation

<400> 169

Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val

1 5 10 15

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

20 25 30

Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu

35 40 45

Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

50 55 60

Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser

65 70 75 80

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

85 90 95

Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile

100 105 110

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

115 120 125

Pro Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Leu

130 135 140

Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val

145 150 155 160

Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro

165 170 175

Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr

180 185 190

Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val

195 200 205

Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu

210 215 220

Ser Pro Gly

225

<210> 170

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-172-006 without LALA mutation

<400> 170

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgcgtaaata ctacccgccg 420

tacaaccagc tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgg gcgcagatag gtggctggaa 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 171

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-006/HelD1.3 with LALA mutation

<400> 171

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Leu Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 172

<211> 450

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain of mimic MAT2 FS22-172-006/HelD1.3 without LALA mutation

<400> 172

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Arg Lys Tyr Tyr Pro Pro Tyr Asn Gln Leu Ser

355 360 365

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

370 375 380

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

385 390 395 400

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

405 410 415

Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser Val Met

420 425 430

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

435 440 445

Pro Gly

450

<210> 173

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain of the imitation MAT2 HelD1.3

<400> 173

Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Gly Asn Ile His Asn Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val

35 40 45

Tyr Asn Ala Lys Thr Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Gly Ser Tyr Tyr Cys Gln His Phe Trp Ser Thr Pro Arg

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 174

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 domain structural loop sequence of Fcab FS22-053

<400> 174

Tyr Tyr Asn Arg Trp Gln Asp

1 5

<210> 175

<211> 111

<212> Protein

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053

<400> 175

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp

1 5 10 15

Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln Val Ser Leu Thr Cys

20 25 30

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

35 40 45

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

50 55 60

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Tyr Tyr Asn

65 70 75 80

Arg Trp Gln Asp Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

85 90 95

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105 110

<210> 176

<211> 333

<212> DNA

<213> Artificial sequence

<220>

<223> CH3 Domain Fcab FS22-053

<400> 176

ggccagcctc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaacccg 60

ccgtacctgt tctctaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 120

gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 180

cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt gtattataac 240

aggtggcagg atgggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 300

tacacacaga agagcctctc cctgtctccg ggt 333

<210> 177

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053 Fcab with LALA

<400> 177

acttgcccgc cttgcccagc cccggaagct gccggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgt attataacag gtggcaggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 178

<211> 681

<212> DNA

<213> Artificial sequence

<220>

<223> Fcab FS22-053 Fcab without LALA mutation

<400> 178

acttgcccgc cttgcccagc cccggaactg ctgggtggtc cttcggtgtt cctcttcccg 60

cccaagccga aggataccct gatgatctca cggacccccg aagtgacctg tgtggtggtg 120

gacgtgtccc acgaggaccc ggaagtgaaa ttcaattggt acgtggatgg agtggaagtg 180

cacaacgcca agaccaagcc acgggaagaa cagtacaact ctacctaccg cgtggtgtcc 240

gtgctcactg tgctgcacca agactggctg aacgggaagg agtacaagtg caaagtgtcc 300

aacaaggcgc tgcctgcccc aattgagaaa actatctcga aagccaaggg ccagcctcga 360

gaaccacagg tgtacaccct gcccccatcc cgggatgagc tgaacccgcc gtacctgttc 420

tctaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 480

gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 540

tccgacggct ccttcttcct ctacagcaag ctcaccgtgt attataacag gtggcaggat 600

gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacacagaag 660

agcctctccc tgtctccggg t 681

<210> 179

<211> 15

<212> Protein

<213> Artificial sequence

<220>

<223> Full-length hinge region of immunoglobulin

<400> 179

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

1 5 10 15

<210> 180

<211> 232

<212> Protein

<213> CD137 Homo sapiens

<400> 180

Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn

1 5 10 15

Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser

20 25 30

Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val

35 40 45

Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp

50 55 60

Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu

65 70 75 80

Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp

85 90 95

Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr

115 120 125

Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro

130 135 140

Gly Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His

145 150 155 160

Ser Pro Gln Ile Ile Ser Phe Phe Leu Ala Leu Thr Ser Thr Ala Leu

165 170 175

Leu Phe Leu Leu Phe Phe Leu Thr Leu Arg Phe Ser Val Val Lys Arg

180 185 190

Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro

195 200 205

Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu

210 215 220

Glu Glu Glu Gly Gly Cys Glu Leu

225 230

<210> 181

<211> 163

<212> Protein

<213> Homo sapiens CD137 extracellular domain

<400> 181

Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn

1 5 10 15

Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser

20 25 30

Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val

35 40 45

Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp

50 55 60

Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu

65 70 75 80

Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp

85 90 95

Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr

115 120 125

Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro

130 135 140

Gly Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His

145 150 155 160

Ser Pro Gln

<210> 182

<211> 231

<212> Protein

<213> CD137 cynomolgus macaque

<400> 182

Leu Gln Asp Leu Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn

1 5 10 15

Asn Arg Ser Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser

20 25 30

Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val

35 40 45

Phe Lys Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp

50 55 60

Cys Ile Ser Gly Tyr His Cys Leu Gly Ala Glu Cys Ser Met Cys Glu

65 70 75 80

Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp

85 90 95

Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr

115 120 125

Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro

130 135 140

Gly Ala Ser Ser Ala Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His

145 150 155 160

Ser Pro Gln Ile Ile Phe Phe Leu Ala Leu Thr Ser Thr Val Val Leu

165 170 175

Phe Leu Leu Phe Phe Leu Val Leu Arg Phe Ser Val Val Lys Arg Ser

180 185 190

Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val

195 200 205

Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu

210 215 220

Glu Glu Gly Gly Cys Glu Leu

225 230

<210> 183

<211> 163

<212> Protein

<213> Extracellular domain of CD137 from cynomolgus monkey

<400> 183

Leu Gln Asp Leu Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn

1 5 10 15

Asn Arg Ser Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser

20 25 30

Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val

35 40 45

Phe Lys Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp

50 55 60

Cys Ile Ser Gly Tyr His Cys Leu Gly Ala Glu Cys Ser Met Cys Glu

65 70 75 80

Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp

85 90 95

Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr

115 120 125

Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro

130 135 140

Gly Ala Ser Ser Ala Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His

145 150 155 160

Ser Pro Gln

<210> 184

<211> 233

<212> Protein

<213> CD137 mouse

<400> 184

Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys

1 5 10 15

Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile

20 25 30

Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe

35 40 45

Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys

50 55 60

Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys

65 70 75 80

Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys Thr Cys

85 90 95

Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr Gly Thr

115 120 125

Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe Ser Pro

130 135 140

Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly His Ser

145 150 155 160

Leu Gln Val Leu Thr Leu Phe Leu Ala Leu Thr Ser Ala Leu Leu Leu

165 170 175

Ala Leu Ile Phe Ile Thr Leu Leu Phe Ser Val Leu Lys Trp Ile Arg

180 185 190

Lys Lys Phe Pro His Ile Phe Lys Gln Pro Phe Lys Lys Thr Thr Gly

195 200 205

Ala Ala Gln Glu Glu Asp Ala Cys Ser Cys Arg Cys Pro Gln Glu Glu

210 215 220

Glu Gly Gly Gly Gly Gly Tyr Glu Leu

225 230

<210> 185

<211> 164

<212> Protein

<213> Mouse CD137 extracellular domain

<400> 185

Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys

1 5 10 15

Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile

20 25 30

Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe

35 40 45

Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys

50 55 60

Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys

65 70 75 80

Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys Thr Cys

85 90 95

Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys Arg Pro

100 105 110

Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr Gly Thr

115 120 125

Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe Ser Pro

130 135 140

Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly His Ser

145 150 155 160

Leu Gln Val Leu

<210> 186

<211> 445

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain MAT G1/HelD1.3

<400> 186

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 187

<211> 290

<212> Protein

<213> PD-L1 Homo sapiens

<400> 187

Met Arg Ile Phe Ala Val Phe Ile Phe Met Thr Tyr Trp His Leu Leu

1 5 10 15

Asn Ala Phe Thr Val Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr

20 25 30

Gly Ser Asn Met Thr Ile Glu Cys Lys Phe Pro Val Glu Lys Gln Leu

35 40 45

Asp Leu Ala Ala Leu Ile Val Tyr Trp Glu Met Glu Asp Lys Asn Ile

50 55 60

Ile Gln Phe Val His Gly Glu Glu Asp Leu Lys Val Gln His Ser Ser

65 70 75 80

Tyr Arg Gln Arg Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn

85 90 95

Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr

100 105 110

Arg Cys Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Val

115 120 125

Lys Val Asn Ala Pro Tyr Asn Lys Ile Asn Gln Arg Ile Leu Val Val

130 135 140

Asp Pro Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr

145 150 155 160

Pro Lys Ala Glu Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser

165 170 175

Gly Lys Thr Thr Thr Asn Ser Lys Arg Glu Glu Lys Leu Phe Asn

180 185 190

Val Thr Ser Thr Leu Arg Ile Asn Thr Thr Thr Asn Glu Ile Phe Tyr

195 200 205

Cys Thr Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu Leu

210 215 220

Val Ile Pro Glu Leu Pro Leu Ala His Pro Pro Asn Glu Arg Thr His

225 230 235 240

Leu Val Ile Leu Gly Ala Ile Leu Leu Cys Leu Gly Val Ala Leu Thr

245 250 255

Phe Ile Phe Arg Leu Arg Lys Gly Arg Met Met Asp Val Lys Lys Cys

260 265 270

Gly Ile Gln Asp Thr Asn Ser Lys Lys Gln Ser Asp Thr His Leu Glu

275 280 285

GluThr

290

<210> 188

<211> 290

<212> Protein

<213> PD-L1 mice

<400> 188

Met Arg Ile Phe Ala Gly Ile Ile Phe Thr Ala Cys Cys His Leu Leu

1 5 10 15

Arg Ala Phe Thr Ile Thr Ala Pro Lys Asp Leu Tyr Val Val Glu Tyr

20 25 30

Gly Ser Asn Val Thr Met Glu Cys Arg Phe Pro Val Glu Arg Glu Leu

35 40 45

Asp Leu Leu Ala Leu Val Val Tyr Trp Glu Lys Glu Asp Glu Gln Val

50 55 60

Ile Gln Phe Val Ala Gly Glu Glu Asp Leu Lys Pro Gln His Ser Asn

65 70 75 80

Phe Arg Gly Arg Ala Ser Leu Pro Lys Asp Gln Leu Leu Lys Gly Asn

85 90 95

Ala Ala Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr

100 105 110

Cys Cys Ile Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Leu

115 120 125

Lys Val Asn Ala Pro Tyr Arg Lys Ile Asn Gln Arg Ile Ser Val Asp

130 135 140

Pro Ala Thr Ser Glu His Glu Leu Ile Cys Gln Ala Glu Gly Tyr Pro

145 150 155 160

Glu Ala Glu Val Ile Trp Thr Asn Ser Asp His Gln Pro Val Ser Gly

165 170 175

Lys Arg Ser Val Thr Thr Ser Arg Thr Glu Gly Met Leu Leu Asn Val

180 185 190

Thr Ser Ser Leu Arg Val Asn Ala Thr Ala Asn Asp Val Phe Tyr Cys

195 200 205

Thr Phe Trp Arg Ser Gln Pro Gly Gln Asn His Thr Ala Glu Leu Ile

210 215 220

Ile Pro Glu Leu Pro Ala Thr His Pro Pro Gln Asn Arg Thr His Trp

225 230 235 240

Val Leu Leu Gly Ser Ile Leu Leu Phe Leu Ile Val Val Ser Thr Val

245 250 255

Leu Leu Phe Leu Arg Lys Gln Val Arg Met Leu Asp Val Glu Lys Cys

260 265 270

Gly Val Glu Asp Thr Ser Ser Lys Asn Arg Asn Asp Thr Gln Phe Glu

275 280 285

GluThr

290

<210> 189

<211> 303

<212> Protein

<213> Mouse mesothelin

<400> 189

Asp Ala Glu Gln Lys Ala Cys Pro Pro Gly Lys Glu Pro Tyr Lys Val

1 5 10 15

Asp Glu Asp Leu Ile Phe Tyr Gln Asn Trp Glu Leu Glu Ala Cys Val

20 25 30

Asp Gly Thr Met Leu Ala Arg Gln Met Asp Leu Val Asn Glu Ile Pro

35 40 45

Phe Thr Tyr Glu Gln Leu Ser Ile Phe Lys His Lys Leu Asp Lys Thr

50 55 60

Tyr Pro Gln Gly Tyr Pro Glu Ser Leu Ile Gln Gln Leu Gly His Phe

65 70 75 80

Phe Arg Tyr Val Ser Pro Glu Asp Ile His Gln Trp Asn Val Thr Ser

85 90 95

Pro Asp Thr Val Lys Thr Leu Leu Lys Val Ser Lys Gly Gln Lys Met

100 105 110

Asn Ala Gln Ala Ile Ala Leu Val Ala Cys Tyr Leu Arg Gly Gly Gly

115 120 125

Gln Leu Asp Glu Asp Met Val Lys Ala Leu Gly Asp Ile Pro Leu Ser

130 135 140

Tyr Leu Cys Asp Phe Ser Pro Gln Asp Leu His Ser Val Pro Ser Ser

145 150 155 160

Val Met Trp Leu Val Gly Pro Gln Asp Leu Asp Lys Cys Ser Gln Arg

165 170 175

His Leu Gly Leu Leu Tyr Gln Lys Ala Cys Ser Ala Phe Gln Asn Val

180 185 190

Ser Gly Leu Glu Tyr Phe Glu Lys Ile Lys Thr Phe Leu Gly Gly Ala

195 200 205

Ser Val Lys Asp Leu Arg Ala Leu Ser Gln His Asn Val Ser Met Asp

210 215 220

Ile Ala Thr Phe Lys Arg Leu Gln Val Asp Ser Leu Val Gly Leu Ser

225 230 235 240

Val Ala Glu Val Gln Lys Leu Leu Gly Pro Asn Ile Val Asp Leu Lys

245 250 255

Thr Glu Glu Asp Lys Ser Pro Val Arg Asp Trp Leu Phe Arg Gln His

260 265 270

Gln Lys Asp Leu Asp Arg Leu Gly Leu Gly Leu Gln Gly Gly Ile Pro

275 280 285

Asn Gly Tyr Leu Val Leu Asp Phe Asn Val Arg Glu Ala Phe Ser

290 295 300

<210> 190

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain MAT2 FS22m-063-AA/FS28m-228

<400> 190

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Phe Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Met Ile Ser Pro Lys Ser Ser Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Trp Phe Thr Pro Ala Arg Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Pro Tyr Trp Ser Tyr Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Met Asn Tyr

405 410 415

Arg Trp Glu Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 191

<211> 215

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain MAT2 FS22m-063-AA/FS28m-228

<400> 191

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Pro Phe Pro Phe Ser

85 90 95

Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala

100 105 110

Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser

115 120 125

Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu

130 135 140

Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser

145 150 155 160

Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu

165 170 175

Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val

180 185 190

Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys

195 200 205

Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 192

<211> 445

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain MAT G1AA/HelD1.3 (with LALA)

<400> 192

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ser Thr Phe Ser Gly Tyr

20 25 30

Gly Val Asn Trp Val Arg Gln Pro Pro Gly Arg Gly Leu Glu Trp Ile

35 40 45

Gly Met Ile Trp Gly Asp Gly Asn Thr Asp Tyr Asn Ser Ala Leu Lys

50 55 60

Ser Arg Val Thr Met Leu Val Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Arg Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Arg Glu Arg Asp Tyr Arg Leu Asp Tyr Trp Gly Gln Gly Ser Leu Val

100 105 110

Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala

115 120 125

Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu

130 135 140

Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly

145 150 155 160

Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser

165 170 175

Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu

180 185 190

Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr

195 200 205

Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val

260 265 270

Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 193

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-004-AA/S70 (with LALA)

<400> 193

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 194

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22-172-004-AA/S70 (no LALA mutation)

<400> 194

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Tyr Ile Tyr Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 195

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-003-AA/S70 (with LALA)

<400> 195

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 196

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22-172-003-AA/S70 (no LALA mutation)

<400> 196

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Tyr Ile Ile Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 197

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-002-AA/S70 (with LALA)

<400> 197

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 198

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22-172-002-AA/S70 (no LALA mutation)

<400> 198

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 199

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-002-AA/S70

<400> 199

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 200

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-002-AA/S70 (with LALA)

<400> 200

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Pro Phe Gln Met Pro Pro Tyr Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 201

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22-053-011-AA/S70 (no LALA mutation)

<400> 201

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 202

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-053-011-AA/S70 (with LALA)

<400> 202

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Asp Tyr Trp Arg Trp Thr Asp Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 203

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-053-008-AA/S70 (with LALA)

<400> 203

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 204

<211> 452

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22-053-008-AA/S70 (no LALA mutation)

<400> 204

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Trp Ile Ser Pro Tyr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Ala Asp Thr Ser Lys Asn Thr Ala Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg His Trp Pro Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr Leu Phe Ser Asn Gln

355 360 365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375 380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395 400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410 415

Thr Val Glu His Thr Arg Trp Leu Asp Gly Asn Val Phe Ser Cys Ser

420 425 430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440 445

Leu Ser Pro Gly

450

<210> 205

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22m-063-AA/F2 (no LALA mutation)

<400> 205

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Glu Asn Trp Gly Ser Tyr Phe Asp Leu Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Pro Tyr Trp Ser Tyr Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Met Asn Tyr

405 410 415

Arg Trp Glu Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 206

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22m-063-AA/F2 (no LALA mutation)

<400> 206

Glu Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Lys Glu Asn Trp Gly Ser Tyr Phe Asp Leu Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Pro Tyr Trp Ser Tyr Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Met Asn Tyr

405 410 415

Arg Trp Glu Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 207

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain FS22m-063-AA/F2

<400> 207

Asp Ile Val Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Arg Ala Pro Lys Val Leu Ile

35 40 45

Tyr Lys Ala Ser Thr Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 208

<211> 447

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy chain FS22m-063/FS28m-228 (no LALA mutation)

<400> 208

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr

20 25 30

Phe Met Val Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Met Ile Ser Pro Lys Ser Ser Asn Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Trp Phe Thr Pro Ala Arg Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Asp Glu Pro Tyr Trp Ser Tyr Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Met Asn Tyr

405 410 415

Arg Trp Glu Leu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

<210> 209

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain FS22-172-004-AA/S70

<400> 209

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Val Ser Thr Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ser Ala Ser Phe Leu Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Tyr His Pro Ala

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 210

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR1 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 210

Gly Tyr Pro Phe Thr Ser Tyr Gly

1 5

<210> 211

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR1 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 211

Ser Tyr Gly Ile Ser

1 5

<210> 212

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR2 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 212

Ile Ser Ala Tyr Ser Gly Gly Thr

1 5

<210> 213

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR2 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 213

Trp Ile Ser Ala Tyr Ser Gly Gly Thr Asn Tyr Ala Gln Lys Leu Gln

1 5 10 15

Gly

<210> 214

<211> 16

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR3 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 214

Ala Arg Asp Leu Phe Pro Thr Ile Phe Gly Val Ser Tyr Tyr Tyr Tyr

1 5 10 15

<210> 215

<211> 14

<212> Protein

<213> Artificial sequence

<220>

<223> HCDR3 VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 215

Asp Leu Phe Pro Thr Ile Phe Gly Val Ser Tyr Tyr Tyr Tyr

1 5 10

<210> 216

<211> 123

<212> Protein

<213> Artificial sequence

<220>

<223> VH domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

<400> 216

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Pro Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Ser Gly Gly Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Leu Phe Pro Thr Ile Phe Gly Val Ser Tyr Tyr Tyr Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 217

<211> 6

<212> Protein

<213> Artificial sequence

<220>

<223> LCDRR1 VL domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 217

Gln Ser Ile Gly Asn Arg

1 5

<210> 218

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR1 VL domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 218

Arg Ala Ser Gln Ser Ile Gly Asn Arg Leu Ala

1 5 10

<210> 219

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR2 domain VL FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 219

Glu Ala Ser

1

<210> 220

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR2 domain VL FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 220

Glu Ala Ser Thr Ser Glu Thr

1 5

<210> 221

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR3 domain VL FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (IMGT)

<400> 221

Gln Gln Ser Tyr Ser Thr Pro Tyr Thr

1 5

<210> 222

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR3 domain VL FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2 (Kabat)

<400> 222

Gln Gln Ser Tyr Ser Thr Pro Tyr Thr

1 5

<210> 223

<211> 107

<212> Protein

<213> Artificial sequence

<220>

<223> VL domain FS22-172-003-AA/E12v2 and FS22-053-008-AA/E12v2

<400> 223

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Arg

1 5 10 15

Asp Arg Val Ile Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly Asn Arg

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Glu Ala Ser Thr Ser Glu Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 224

<211> 457

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-172-003-AA/E12v2

<400> 224

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Pro Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Ser Gly Gly Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Leu Phe Pro Thr Ile Phe Gly Val Ser Tyr Tyr Tyr Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

115 120 125

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

130 135 140

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

145 150 155 160

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

165 170 175

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

180 185 190

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

195 200 205

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys

210 215 220

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

225 230 235 240

Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

325 330 335

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Pro Tyr Ile Ile

355 360 365

Pro Pro Tyr Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

370 375 380

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

385 390 395 400

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

405 410 415

Leu Tyr Ser Lys Leu Thr Val Gly Ala Asp Arg Trp Leu Glu Gly Asn

420 425 430

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

435 440 445

Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455

<210> 225

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain FS22-172-003-AA/E12v2 and light chain FS22-053-008-AA/E12v2

<400> 225

Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Arg

1 5 10 15

Asp Arg Val Ile Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly Asn Arg

20 25 30

Leu Ala Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Glu Ala Ser Thr Ser Glu Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 226

<211> 457

<212> Protein

<213> Artificial sequence

<220>

<223> Heavy Chain FS22-053-008-AA/E12v2

<400> 226

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Pro Phe Thr Ser Tyr

20 25 30

Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Ser Ala Tyr Ser Gly Gly Thr Asn Tyr Ala Gln Lys Leu

50 55 60

Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Leu Phe Pro Thr Ile Phe Gly Val Ser Tyr Tyr Tyr Tyr

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

115 120 125

Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

130 135 140

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

145 150 155 160

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

165 170 175

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

180 185 190

Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

195 200 205

Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys

210 215 220

Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

225 230 235 240

Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

245 250 255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265 270

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

275 280 285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

290 295 300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315 320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

325 330 335

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345 350

Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Asn Pro Pro Tyr

355 360 365

Leu Phe Ser Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr

370 375 380

Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

385 390 395 400

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

405 410 415

Leu Tyr Ser Lys Leu Thr Val Asp Tyr Trp Arg Trp Leu Glu Gly Asn

420 425 430

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

435 440 445

Gln Lys Ser Leu Ser Leu Ser Pro Gly

450 455

<210> 227

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<223> VH domain CDR1 FS28-256-271 (IMGT)

<400> 227

Gly Phe Thr Phe Thr His Thr Tyr

1 5

<210> 228

<211> 5

<212> Protein

<213> Artificial sequence

<220>

<223> CDR1 of VH domain FS28-256-271 (Kabat)

<400> 228

His Thr Tyr Met Ser

1 5

<210> 229

<211> 8

<212> Protein

<213> Artificial sequence

<220>

<223> VH domain CDR2 FS28-256-271 (IMGT)

<400> 229

Ile Ser Pro Thr Tyr Ser Thr Thr

1 5

<210> 230

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<223> CDR2 of VH domain FS28-256-271 (Kabat)

<400> 230

Ala Ile Ser Pro Thr Tyr Ser Thr Thr Asn Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 231

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<223> CDR3 of VH domain FS28-256-271 (IMGT)

<400> 231

Ala Arg Tyr Asn Ala Tyr His Ala Ala Leu Asp Tyr

1 5 10

<210> 232

<211> 10

<212> Protein

<213> Artificial sequence

<220>

<223> CDR3 of VH domain FS28-256-271 (Kabat)

<400> 232

Tyr Asn Ala Tyr His Ala Ala Leu Asp Tyr

1 5 10

<210> 233

<211> 119

<212> Protein

<213> Artificial sequence

<220>

<223> Domain VH FS28-256-271

<400> 233

Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr His Thr

20 25 30

Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Asn Ile Ser Pro Thr Tyr Ser Thr Thr Asn Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Asn Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Tyr Asn Ala Tyr His Ala Ala Leu Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 234

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR1 domain VL FS28-256-271 (IMGT)

<400> 234

Gln Ser Val Ser Ser Ser Tyr

1 5

<210> 235

<211> 12

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR1 domain VL FS28-256-271 (Kabat)

<400> 235

Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala

1 5 10

<210> 236

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR2 domain VL FS28-256-271 (IMGT)

<400> 236

Gly Ala Ser

1

<210> 237

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR2 domain VL FS28-256-271 (Kabat)

<400> 237

Gly Ala Ser Ser Arg Ala Thr

1 5

<210> 238

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR3 domain VL FS28-256-271 (IMGT)

<400> 238

Gln Gln Thr Val Pro Tyr Pro Tyr Thr

1 5

<210> 239

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> LCDR3 domain VL FS28-256-271 (Kabat)

<400> 239

Gln Gln Thr Val Pro Tyr Pro Tyr Thr

1 5

<210> 240

<211> 108

<212> Protein

<213> Artificial sequence

<220>

<223> Domain VL FS28-256-271

<400> 240

Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser

20 25 30

Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu

35 40 45

Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser

50 55 60

Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu

65 70 75 80

Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Thr Val Pro Tyr Pro

85 90 95

Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<---

Claims (57)

1. A specific binding element that binds CD137 and comprises an antigen-binding site for CD137 located in the CH3 domain of said specific binding element, wherein said antigen-binding site for CD137 comprises a first sequence located in the structural loop AB of said CH3 domain, wherein said first sequence comprises the sequence PPY (SEQ ID NO: 10). 2. A specific binding element according to claim 1, characterized in that said specific binding element contains an insert in said structural loop AB. 3. The specific binding element according to claim 2, characterized in that said insert is 5 amino acids in length. 4. A specific binding element according to any one of paragraphs 1-3, characterized in that said first sequence is the first sequence of a specific binding element: (i) FS22-172-003, represented by SEQ ID NO: 138; (ii) FS22-172-002, represented by SEQ ID NO: 129; (iii) FS22-172-004, represented by SEQ ID NO: 147; (iv) FS22-172-001, represented by SEQ ID NO: 120; (v) FS22-172-005, represented by SEQ ID NO: 156; (vi) FS22-172-006, as shown in SEQ ID NO: 110; or (vii) FS22-172, represented by SEQ ID NO: 110. 5. A specific binding element according to any one of claims 1 to 4, characterized in that said specific binding element further comprises a second sequence located in the EF structural loop of said CH3 domain, wherein said second sequence is the second sequence of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, presented in SEQ ID NO: 111. 6. A specific binding element according to any one of paragraphs 1-5, characterized in that said specific binding element contains the sequence of the CH3 domain of a specific binding element: (i) FS22-172-003, represented by SEQ ID NO: 139; (ii) FS22-172-002, represented by SEQ ID NO: 130; (iii) FS22-172-004, represented by SEQ ID NO: 148; (iv) FS22-172-001, represented by SEQ ID NO: 121; (v) FS22-172-005, represented by SEQ ID NO: 157; (vi) FS22-172-006, as represented by SEQ ID NO: 165; or (vii) FS22-172, represented by SEQ ID NO: 112. 7. The specific binding element according to any one of claims 1 to 6, characterized in that said specific binding element comprises the sequence of the specific binding element FS22-172-003, FS22-172-002, FS22-172-004, FS22-172-001, FS22-172-005, FS22-172-006 or FS22-172, presented in SEQ ID NO: 141, 132, 150, 123, 159, 167 and 114, respectively. 8. The specific binding element according to any one of claims 1 to 3, characterized in that said first sequence is the first sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 and FS22-053, presented in SEQ ID NO: 19. 9. A specific binding element according to any one of claims 1-3 or 8, characterized in that said specific binding element additionally comprises a second sequence located in the EF structural loop of said CH3 domain, and wherein said second sequence is the second sequence of the specific binding element: (i) FS22-053-008, represented by SEQ ID NO: 20; (ii) FS22-053-009, represented by SEQ ID NO: 29; (iii) FS22-053-011, represented by SEQ ID NO: 47; (iv) FS22-053-017, represented by SEQ ID NO: 101; (v) FS22-053-014, represented by SEQ ID NO: 74; (vi) FS22-053-010, represented by SEQ ID NO: 38; (vii) FS22-053-012, represented by SEQ ID NO: 56; (viii) FS22-053-013, represented by SEQ ID NO: 65; (ix) FS22-053-015, represented by SEQ ID NO: 83; (x) FS22-053-016, as represented in SEQ ID NO: 92; or (xi) FS22-053, represented by SEQ ID NO: 174. 10. A specific binding element according to any one of paragraphs 1-3 and 8-9, characterized in that said specific binding element contains the sequence of the CH3 domain of a specific binding element: (i) FS22-053-008, represented by SEQ ID NO: 21; (ii) FS22-053-009, represented by SEQ ID NO: 30; (iii) FS22-053-011, represented by SEQ ID NO: 48; (iv) FS22-053-017, represented by SEQ ID NO: 102; (v) FS22-053-014, represented by SEQ ID NO: 75; (vi) FS22-053-010, represented by SEQ ID NO: 39; (vii) FS22-053-012, represented by SEQ ID NO: 57; (viii) FS22-053-013, represented by SEQ ID NO: 66; (ix) FS22-053-015, represented by SEQ ID NO: 84; (x) FS22-053-016, as represented in SEQ ID NO: 93; or (xi) FS22-053, represented by SEQ ID NO: 175. 11. The specific binding element according to any one of claims 1-3 and 8-10, characterized in that said specific binding element comprises the sequence of the specific binding element FS22-053-008, FS22-053-009, FS22-053-011, FS22-053-017, FS22-053-014, FS22-053-010, FS22-053-012, FS22-053-013, FS22-053-015, FS22-053-016 or FS22-053, presented in SEQ ID NO: 23, 32, 50, 104, 77, 41, 59, 68, 86, 95 and 15, respectively. 12. The specific binding element according to any one of claims 1 to 11, characterized in that said specific binding element comprises said first sequence, first and second sequence, CH3 domain sequence, or the sequence of the specific binding element FS22-172-003, FS22-053-008 or FS22-053-017, preferably FS22-172-003 or FS22-053-008, more preferably FS22-172-003. 13. A specific binding element according to any one of claims 1-12, characterized in that said CH3 domain sequence additionally contains a lysine residue (K) immediately at the C-terminus of the CH3 domain sequence. 14. A specific binding element according to any one of claims 1-13, characterized in that said specific binding element further comprises a CDR-based antigen-binding site, optionally wherein said specific binding element is an antibody molecule. 15. The specific binding element of claim 14, wherein said specific binding element is an antibody molecule, and wherein said CDR-based antigen-binding site binds a second antigen selected from the group consisting of: an immune cell antigen, a tumor antigen, and a pathogen antigen. 16. A specific binding element according to any one of claims 1-15, characterized in that said specific binding element is modified to reduce or eliminate binding of said specific binding element to one or more Fcγ receptors. 17. A nucleic acid molecule encoding a specific binding element according to any one of claims 1-16. 18. A recombinant host cell containing the nucleic acid of claim 17, for producing the specific binding element of any one of claims 1-16. 19. A method for obtaining a specific binding element according to any one of claims 1-16, comprising culturing a recombinant host cell according to claim 18 under conditions that ensure the production of said specific binding element. 20. Use of a specific binding element according to any one of claims 1-16 for treating cancer in an individual. 21. Use of a specific binding element according to any one of claims 1-16 for treating an infectious disease in an individual.