RU2807346C2 - Tetravalent symmetrical bispecific antibodies - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: bispecific antibody that binds two different epitopes, a pharmaceutical composition for treating a tumor associated with the first epitope and a second epitope, including the above bispecific antibody, a conjugate that binds the first epitope and the second epitope, a fusion protein that binds the first epitope and the second epitope, a polynucleotide encoding a bispecific antibody, a cell expressing a bispecific antibody, the use of a bispecific antibody or conjugate or fusion protein for the treatment of a tumor associated with a first and second epitope, and the use of a bispecific antibody or conjugate or fusion protein for the preparation of a reagent or kit for detecting a tumor associated with the first epitope and the second epitope.

EFFECT: expanding the scope of agents that bind two different epitopes.

31 cl, 36 dwg, 35 tbl, 6 ex

Description

Field of technology to which the invention relates

The present invention relates to the field of antibodies. In particular, the present invention relates to bispecific antibodies, pharmaceutical compositions containing them and their uses.

Prerequisites for creating the invention

Bispecific antibodies (BsAbs) are antibodies or antibody-like molecules that have two different binding specificities. Bispecific antibodies are widely used in biomedicine, especially in tumor immunotherapy.

Bispecific antibodies can be produced by methods such as chemical engineering, cellular engineering and genetic engineering. An advantage of genetic engineering is that antibodies can be easily modified, so many different forms of bispecific antibody fragments can be designed and produced, including bispecific IgG (BsIgG), complemented IgG, bispecific antibody fragments (BsAb fragments), bispecific fusion proteins, and bispecific conjugates. antibodies (BsAb conjugates), (see Christoph Spiess, Qianting Zhai, Paul J. Carter, Alternative molecular formats and therapeutic applications for bispecific antibodies. Molecular Immunology 67 (2015) 95-106).

First type: bispecific IgG, which have the same structure and molecular weight as natural monoclonal antibodies, but have two different Fabs. (1) In the previously used Triomab technology, rat and mouse hybridomas are again fused to form a “hybridoma” to produce mouse and rat chimeric bispecific antibodies. Purification is carried out in accordance with the selectivity of rat and mouse Fc to the affinity excipient, and then the product of interest is obtained. This technology has disadvantages such as low expressed quantity and low yield after purification. (2) Subsequently, antibody engineering technology, such as leucine zipper, peak-to-trough, or electrostatic interaction, is used to modify the Fc, so that a heterodimer is formed from two different heavy chains, and then the light chain and the heavy chain are paired appropriately using common light chains, peak-to-valley technology or electrostatic interaction, or the desired product of interest is obtained by designing different κ and λ light chain subtypes and purifying using an affinity excipient specific to the κ chain and λ chain. Bispecific antibodies of this type have pharmacokinetics similar to those of natural antibodies, but have the disadvantage that impurities are difficult to identify and remove.

Second type: complemented IgG, where a natural IgG monoclonal antibody is used as a scaffold and an antigen binding domain (such as a variable region fragment (Fv), a single chain variable region fragment (ScFv), an antigen binding fragment (Fab) or a polypeptide, etc.) is fused at the N-terminus or C-terminus of the heavy or light chain of an antibody; bispecific antibodies of this type have a molecular weight of more than 150 kDa and a predominantly symmetrical structure. The pharmacokinetics of augmented IgGs are similar to those of natural antibodies, but augmented IgGs have a longer half-life and are easier to purify. The disadvantages of complemented IgG are low expressed quantity and lack of stability, etc.

Third type: fragments of bispecific antibodies, where fragments of the variable regions of the antibody are retained, and part or even the entire constant region of the antibody is deleted; variable heavy chain (VH) fragments and variable light chain (VL) fragments are typically associated with the formation of ScFv via a linker protein, or spontaneously paired to form an Fv. The molecular weight of bispecific antibodies of this type does not exceed 150 kDa, their half-life is short. The disadvantages of bispecific antibody fragments are as follows: it is difficult to achieve the same level of expression of a bispecific antibody fragment as that of a monoclonal antibody, the molecule is not stable enough, and the yield after purification is low, etc.

Fourth type: bispecific fusion proteins, where fragments of the antibody variable regions are retained and other proteins or pharmaceutical molecules (such as T-cell receptor constant regions, human serum albumin or toxin proteins, etc.) are fused. The molecular weight of this type of bispecific antibodies ranges from 75 kDa to 160 kDa, the purification process is difficult, and molecular stability also has certain risks.

Fifth type: bispecific antibody conjugates, where two different monoclonal antibodies or ScFvs are linked together by chemical methods. The method for producing antibodies of this type is complex and has a low final yield, and therefore antibodies of this type are currently not used essentially anywhere in the world.

All of the above types of bispecific antibodies have some disadvantages, which lead to potential risks in the clinical use of such products. To address these risks, the present invention provides a new bispecific antibody.

The essence of the invention

The present disclosure provides a method for designing and producing a novel bispecific antibody. The specific structure of the bispecific antibody is shown in FIG. 1. All of the antibodies in the present disclosure have a human IgG constant region and refer to complementary IgG or bispecific antibody fragments.

In the present invention, the affinity, stability, biological activity and effectiveness of a new bispecific antibody were tested and compared with three existing bispecific antibodies. The results show that the new bispecific antibody has better stability, biological activity and efficiency, and its expression and production methods are more convenient.

Technical solutions

The bispecific antibody of the present invention has two identical hybrid heavy chains and two identical hybrid light chains, where the two hybrid heavy chains form a pair, and the hybrid light chain and the hybrid heavy chain form a pair.

In some aspects, the hybrid antibody heavy chains are linked by one or more disulfide bonds.

In some aspects, the hybrid light chain and the hybrid antibody heavy chain are linked by one or more disulfide bonds.

In some aspects, the hybrid antibody heavy chain comprises an antibody heavy chain variable region a (VHa), a first constant region (CH1), an antibody light chain variable region b (VLb), and an Fc fragment. Where VLb is located between CH1 and Fc and is connected using a linker or peptide bond. The hybrid antibody light chain comprises an antibody light chain variable region a (VLa), a light chain constant region (CL), and an antibody heavy chain variable region b (VHb). Where VHb is located at the C-terminus of CL and is linked by a linker or peptide bond.

In some aspects, the hybrid antibody heavy chain comprises an antibody heavy chain variable region a (VHa), a first constant region (CH1), an antibody heavy chain variable region b (VHb), and an Fc fragment. Where VHb is located between CH1 and Fc and is connected using a linker or peptide bond. The hybrid antibody light chain comprises an antibody light chain variable region a (VLa), a light chain constant region (CL), and an antibody light chain variable region b (VLb). Where VLb is located at the C-terminus of CL and is linked by a linker or peptide bond.

In some aspects, the VHa-VLa pair is specific for antigen A, which includes, but is not limited to, tumor cell surface antigens, immune cell surface antigens, viruses, bacteria, endotoxins, cytokines such as CD3, SLAMF7, CD38, BCMA, CD16a , CEA, PD-L1, PD-1, CTLA-4, TIGIT, LAG-3, VEGF, B7-H3, TGF-β, IL-10, etc.

In some aspects, the VHb-VLb pair is specific for a B antigen, which includes, but is not limited to, tumor cell surface antigens, immune cell surface antigens, viruses, bacteria, endotoxins, cytokines such as CD3, SLAMF7, CD38, BCMA, CD16a , CEA, PD-L1, PD-1, CTLA-4, TIGIT, LAG-3, VEGF, B7-H3, TGF-β, IL-10, etc.

Specifically, in one aspect, the present invention provides a bispecific antibody comprising two identical hybrid heavy chains and two identical hybrid light chains, wherein the two hybrid heavy chains form a pair, and the hybrid light chain and the hybrid heavy chain form a pair, wherein the hybrid heavy chain includes a variable an antibody a heavy chain region (VHa), a CH1 first constant region, an antibody b variable region 1, and an Fc fragment, and an antibody b variable region 1 linked to CH1 via a linker 2 or a peptide bond, or linked to an Fc via a linker 3 or a peptide bond. The hybrid light chain of a bispecific antibody includes an antibody light chain variable region a (VLa), a light chain constant region CL, and an antibody b variable region 2, wherein antibody b variable region 2 is linked to the C-terminus of CL via linker 1 or a peptide bond, wherein

(1) when antibody b variable region 1 is an antibody b heavy chain variable region (VHb), antibody b variable region 2 is an antibody b light chain variable region (VLb);

or

(2) when antibody b variable region 1 is an antibody light chain variable region b (VLb), antibody b variable region 2 is an antibody heavy chain variable region b (VHb),

where, the VHa-VLa pair targets the A antigen, and the VHb-VLb pair targets the B antigen.

In one embodiment, the structure of the bispecific antibody of the present invention is F(ab) ₂ -(Fv) ₂ -Fc, where F(ab) ₂ includes two VHa, two CH1, two VLa and two CL; (Fv) ₂ includes two VHb and two VLb, where the positions of VHb and VLb are interchangeable; and the Fc fragment includes a hinge region and a second CH2 constant region and a third CH3 constant region.

In one embodiment, the bispecific antibody symmetrically has the structure shown in Formula I from N-terminus to C-terminus:

Where

" " represents a disulfide bond or a covalent bond;

"-" represents a peptide bond;

L1, L2 and L3 each independently represent a peptide bond or linker or hinge;

VHa is the variable region of the heavy chain of antibody a;

VLa is the variable region of the light chain of antibody a;

CH1 represents the first constant region;

Fc is an Fc fragment containing a hinge region;

CL is the light chain constant region;

Vb1 is antibody b variable region 1;

Vb2 is antibody b variable region 2.

In one embodiment, the bispecific antibody symmetrically has the structure shown in Formula II or Formula III from N-terminus to C-terminus:

Where

" " represents a disulfide bond or a covalent bond;

"-" represents a peptide bond;

L1, L2 and L3 are each independently a peptide bond or linker;

VHa is the variable region of the heavy chain of antibody a;

VLa is the variable region of the light chain of antibody a;

CL is the light chain constant region;

H represents the hinge region;

CH1, CH2 and CH3 represent the first constant region, the second constant region and the third constant region, respectively;

VHb is the heavy chain variable region of antibody b;

VLb is the variable region of the light chain of antibody b.

In one embodiment, the bispecific antibody includes:

(1) a hybrid heavy chain, where the hybrid heavy chain includes an antibody heavy chain variable region a (VHa), a first constant region CH1, linker 2 (or peptide bond) and an antibody heavy chain variable region b (VHb), linker 3 (or a peptide bond bond) and Fc fragment in sequence from N-terminus to C-terminus; And

(2) a hybrid light chain, where the hybrid light chain includes an antibody light chain variable region a (VLa), a light chain constant region CL, linker 1 (or peptide bond) and an antibody light chain variable region b (VLb) in sequence from N- end to C-terminus.

In one embodiment, the bispecific antibody includes:

(1) a hybrid heavy chain, where the hybrid heavy chain includes an antibody heavy chain variable region a (VHa), a first constant region CH1, linker 2 (or peptide bond) and an antibody light chain variable region b (VLb), linker 3 (or a peptide bond bond) and Fc fragment in sequence from N-terminus to C-terminus; And

(2) a hybrid light chain, where the hybrid light chain includes an antibody light chain variable region a (VLa), a light chain constant region CL, linker 1 (or peptide bond) and an antibody heavy chain variable region b (VHb) in sequence from N- end to C-terminus.

In one embodiment, the VHa-VLa pair forms one or more interchain disulfide bonds, and the VHb-VLb pair forms one or more interchain disulfide bonds.

In one embodiment, antibody b variable region 1 is an antibody b light chain variable region (VLb), and antibody b variable region 2 is an antibody b heavy chain variable region (VHb).

In one embodiment, linkers 1-3 may be the same or different from each other, and linkers 1-3 are each independently selected from the group consisting of SEQ ID NOs: 69-90, preferably selected from SEQ ID NOs: 71, 72, 73 , 74, 75, 76, 77, 78, 79, 80, 82, 83, 85, 86, 87, 88, 89 or 90.

In one embodiment, the hinge region sequence is selected from the group consisting of SEQ ID NOs: 91-103.

In one embodiment, the CL sequence is selected from the group consisting of SEQ ID NO: 104-110.

In one embodiment, the CH1 sequence is selected from the group consisting of SEQ ID NO: 111-114.

In one embodiment, the CH2 sequence is selected from the group consisting of SEQ ID NO: 115-119.

In one embodiment, the CH3 sequence is selected from the group consisting of SEQ ID NO: 120-128.

In one embodiment, the A antigen and the B antigen may be the same or different from each other, preferably the A antigen or the B antigen are different or represent different epitopes on the same antigen.

In one embodiment, antigen A and antigen B are each independently selected from the group consisting of: immune cell surface antigens, tumor antigens, viruses, bacteria, endotoxins, cytokines, or a combination thereof.

In one embodiment, antigen A and/or antigen B is selected from PD-L1, PD-1, VEGFA, IL-10, IL-10R, BCMA, VEGF, TGF-β, CTLA-4, LAG-3, TIGIT, CEA , CD38, SLAMF7, B7-H3, Her2, EpCAM, CD19, CD20, CD30, CD33, CD47, CD52, CD133, EGFR, GD2, GD3, GM2, RANKL, CD3 and/or CD16a.

In one embodiment, antigen A and/or antigen B are selected from SEQ ID NO: 129-145.

In one embodiment, the A antigen and/or B antigen is PD-1; preferably, one of antigen A and antigen B is PD-1 and the other is selected from the group consisting of PD-L1, PD-1, VEGFA, IL-10, IL-10R, BCMA, VEGF, TGF-β, CTLA -4, LAG-3, TIGIT, CEA, CD38, SLAMF7, B7-H3, HER2, CD3 or CD16a.

In one embodiment, the A antigen and the B antigen are selected from the group consisting of:

PD-L1 and VEGF,

PD-1 and VEGF,

PD-L1 and TGF-β,

PD-1 and TGF-β,

PD-1 and CTLA-4,

PD-1 and LAG-3,

PD-1 and TIGIT,

PD-1 and IL-10,

SLAMF7 and CD16a, and

Her2 and Her2.

In one embodiment, VHa, VLa, VHb and/or VLb are derived from an antibody selected from the group consisting of antibodies of animal origin (such as murine antibodies), chimeric antibodies and humanized antibodies; preferably humanized antibodies include fully humanized antibodies and partially humanized antibodies.

In one embodiment, VHa and/or VHb include any of the following sequences:

a) the amino acid sequence shown in any of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37 , 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 146, 148;

b) an amino acid sequence having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the amino acid sequence of a );

c) an amino acid sequence that has one or more (preferably one or more, more preferably 1, 2 or 3) amino acid modifications compared to the amino acid sequence of a); and/or

VLa and/or VLb include any of the following sequences:

d) the amino acid sequence shown in any of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38 , 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 147, 149;

e) an amino acid sequence having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the amino acid sequence of d );

f) an amino acid sequence that has one or more (preferably one or more, more preferably 1, 2, or 3) amino acid modifications compared to the amino acid sequence of d).

In one embodiment, the bispecific antibody is selected from the group consisting of Y100-A1, Y100-A2, Y100-A3, Y100-A4, Y100-A5, Y100-A6, Y100-A7, Y100-A8, Y100-A9, Y100- A10, Y100-A11, Y100-AC1, Y100-AC2, Y100-AC3, Y101-A1, Y101-A2, Y101-A3, Y101-A4, Y101-A5, Y103-A1, Y103-A2, Y103-A3, Y104-A1, Y104-A2, Y104-A3, Y105-A1, Y105-A2, Y105-A3, Y106-A1, Y106-A2, Y106-A3, Y106-A4, Y106-A5, Y110-A1, Y110- A2, Y110-A3, Y110-A4, Y116-A1, Y116-A2, Y116-A3, Y100-B1, Y100-B2, Y100-B3, Y100-B4, Y100-B5, Y100-B6, Y100-B7, Y100-B8, Y100-B9, Y100-B10, Y100-B11, Y100-B12, Y100-B13, Y100-BC1, Y100-BC2, Y100-BC3, Y101-B1, Y101-B2, Y101-B3, Y101- B4, Y101-B5, Y101-B6, Y103-B1, Y103-B2, Y103-B3, Y104-B1, Y104-B2, Y104-B3, Y105-B1, Y105-B2, Y105-B3, Y106-B1, Y106-B2, Y106-B3, Y106-B4, Y106-B5, Y110-B1, Y110-B2, Y110-B3, Y110-B4, Y110-B5, Y116-B1, Y116-B2, Y116-B3, Y140- A1, Y140-A2, Y140-A3, Y140-A4, Y140-B1, Y140-B2, Y140-B3, Y140-B4, or combinations thereof.

In one embodiment, the bispecific antibody is selected from those shown in any of Table 27 and Table 34.

In one embodiment, the bispecific antibody is selected from the group consisting of Y100-B6, Y100-B7, Y100-B8, Y100-B9, Y100-B10, Y100-B11, Y100-B12, Y100-BC1, Y100-BC2, Y100- BC3, Y101-B1, Y101-B2, Y101-B3, Y101-B4, Y101-B5, Y101-B6, Y103-B1, Y103-B2, Y103-B3, Y104-B1, Y104-B2 , Y104-B3, Y105-B1, Y105-B2, Y105-B3, Y106-B1, Y106-B2, Y106-B3, Y106-B4, Y106-B5, Y110-B1, Y110-B2, Y110-B3, Y110-B4, Y110- B5, Y116-B1, Y116-B2, Y116-B3, Y140-B1, Y140-B2, Y140-B3, Y140-B4, or combinations thereof.

In one embodiment, the bispecific antibody is selected from the group consisting of Y100-B7, Y100-B9, Y101-B2, Y101-B4, Y103-B2, Y104-B2, Y105-B2, Y106-B2, Y106-B4, Y110- B2, Y110-B4, Y116-B2, or combinations thereof.

In one embodiment, the bispecific antibody is selected from the group consisting of the following:

(1) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 82 and SEQ ID NO: 42; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 82, SEQ ID NO: 41, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(2) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 42; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 41, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(3) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 42; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 41, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(4) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 46; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 45, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(5) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 46; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 45, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(6) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(7) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(8) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(9) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(10) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 54; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 53, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(11) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(12) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 68; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 67, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(13) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(14) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(15) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(16) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(17) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(18) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(19) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 34; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 33, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(20) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 34; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 33, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(21) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 34; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 33, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(22) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 28; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 27, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(23) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 28; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 27, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(24) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 28; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 27, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(25) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 30; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 29, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(26) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 30; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 29, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(27) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 30; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 29, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(28) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(29) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(30) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(31) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(32) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(33) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(34) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(35) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(36) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(37) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 48; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 47, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(38) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 48; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 47, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(39) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 48; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 47, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(40) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 82 and SEQ ID NO: 41; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 82, SEQ ID NO: 42, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(41) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 41; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 42, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(42) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 41; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 42, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(43) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 45; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 46, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(44) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 45; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 46, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(45) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(46) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(47) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(48) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(49) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 53; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 54, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(50) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 84 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 84, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(51) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 86 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 86, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(52) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(53) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 67; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 68, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(54) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(55) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(56) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(57) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(58) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(59) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(60) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 84 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 84, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(61) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 86 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 86, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(62) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 33; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 34, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(63) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 86 and SEQ ID NO: 33; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 86, SEQ ID NO: 34, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(64) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 86 and SEQ ID NO: 33; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 86, SEQ ID NO: 34, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(65) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 27; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 28, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(66) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 27; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 28, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(67) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 27; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 28, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(68) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 29; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 30, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(69) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 29; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 30, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(70) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 29; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 30, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(71) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(72) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(73) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(74) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(75) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(76) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(77) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(78) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(79) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(80) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(80) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 89 and SEQ ID NO: 47; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 83, SEQ ID NO: 48, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(81) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 47; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 48, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(82) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 47; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 48, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(83) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 149; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 148, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(84) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 147; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 146, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(85) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 149; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 148, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(86) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 147; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 146, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120;

(87) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 148; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 149, SEQ ID NO: 87, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(88) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 146; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 147, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120;

(89) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 148; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 149, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; or

(90) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 146; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 147, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120.

In one embodiment, the bispecific antibody has simultaneous binding activity to an A antigen and a B antigen.

In one embodiment, the bispecific antibody is a biparatope antibody.

In another aspect, the present invention provides a conjugate or fusion protein comprising the bispecific antibodies of the present invention.

In a specific embodiment, the conjugate or fusion protein includes substance A conjugated or fused to an antibody, and substance A is selected from the group consisting of therapeutic agents, prodrugs, proteins (such as enzymes), viruses, lipids, biological response modifiers (such as immunomodulators ), PEGs, hormones, oligonucleotides, diagnostic agents, cytotoxic agents which may be drugs or toxins, ultrasound enhancing agents, non-radioactive markers, detectable markers such as chemiluminescent labeling compounds (such as luminol, isoluminol, thermoluminescent acridinium ester, imidazole , acridinium salt and oxalate), or fluorescent emitting metals (such as 152Eu, or lanthanide tags).

In one embodiment, the conjugate or fusion protein is a monomer, dimer, or multimer.

In another aspect, the present invention provides a pharmaceutical composition comprising a bispecific antibody of the present invention or a conjugate or fusion protein of the present invention.

In one embodiment, the pharmaceutical composition also includes a pharmaceutically acceptable carrier.

In one embodiment, the dosage form of the pharmaceutical composition includes a dosage form for gastrointestinal administration or a dosage form for parenteral administration; preferably, the dosage form of the pharmaceutical composition is an injection, including intravenous injection, intravenous infusion, subcutaneous injection, local injection, intramuscular injection, intratumoral injection, intraperitoneal injection, intracranial injection or intracavitary injection.

In yet another aspect, the present invention provides a polynucleotide encoding a bispecific antibody of the present invention.

In one embodiment, the polynucleotide comprises a first polynucleotide encoding a hybrid light chain of a bispecific antibody and a second polynucleotide encoding a hybrid heavy chain. Preferably, the ratio of the first polynucleotide to the second polynucleotide is 1:1.

In another aspect, the present invention provides a vector comprising a polynucleotide of the present invention; preferably the vector includes: plasmids, phages, yeast plasmids, plant viruses, mammalian viruses such as adenoviruses, adeno-associated viruses, retroviruses or a combination thereof.

In yet another aspect, the present invention provides a cell comprising a polynucleotide of the invention.

In one embodiment, the cell includes a vector of the present invention, or the genome of the cell is integrated with a polynucleotide of the present invention.

In one embodiment, the cell is selected from the group consisting of Escherichia coli , Bacillus subtilis , yeast cells, insect cells, mammalian cells, or a combination thereof; preferably is a mammalian cell, such as a CHO-S cell or a 293E cell.

In yet another aspect, the present invention provides the use of a bispecific antibody or conjugate or fusion protein of the present invention for the treatment of tumors (cancer) or for the preparation of drugs for the treatment of tumors (cancer).

In one embodiment, the disease is a cancer or tumor. Preferably, the tumor is selected from the group consisting of hematological tumors, solid tumors, or a combination thereof.

In another aspect, the present invention provides the use of a bispecific antibody or conjugate or fusion protein of the present invention for the preparation of tumor (cancer) detection reagents or kits.

In yet another aspect, the present invention provides a method for producing a bispecific antibody of the present invention, comprising the following steps:

(i) culturing the cells of the present invention under suitable conditions to obtain a mixture containing the bispecific antibody of the present invention; And

(ii) purifying and/or separating the mixture obtained in step (i) to obtain the bispecific antibody of the present invention.

In one embodiment, the purification includes: affinity chromatography, ion exchange chromatography, hydrophobic interaction chromatography, molecular sieve chromatography, or a combination thereof.

In another aspect, the present invention provides a method for treating diseases, comprising the steps of: administering a therapeutically effective amount of a bispecific antibody of the present invention, or a conjugate or fusion protein of the present invention, or a pharmaceutical composition of the present invention, or a combination thereof, to a subject in need thereof.

In one embodiment, the subject is a human or non-human mammal.

Brief description of drawings

Fig. 1 schematically represents the structure of an antibody having bispecific antibody structure 1 (A), and schematically represents the primary protein structure of each component of the antibody (B), wherein antibody b variable region 1 is represented by VL/Hb and antibody b variable region 2 is represented by VH/ Lb.

Fig. 2 schematically shows the structure of an antibody having a bispecific antibody structure 2 (A), and schematically shows the primary protein structure of each component of the antibody (B).

Fig. 3 schematically shows the structure of an antibody having a bispecific antibody structure 3 (A), and schematically shows the primary protein structure of each component of the antibody (B).

Fig. 4 schematically shows the structure of an antibody having a bispecific antibody structure 4 (A), and schematically shows the primary protein structure of each component of the antibody (B).

Fig. 5 shows antibodies with bispecific antibody structure 1. (A) shows the expression level of Y100-A series and Y100-B series antibodies upon transient transfection in CHO cells. (B) shows the expression levels of Y101/Y103/Y104/Y105-A series and Y101/Y103/Y104/Y105-B series antibodies upon transient transfection in CHO cells. (C) shows the expression level of Y106/Y110/Y116-A series and Y106/Y110/Y116-B series antibodies upon transient transfection in CHO cells. Fig. 5D shows the purity determined by size exclusion HPLC after protein A affinity chromatography for Y100/Y101/Y103/Y104/Y105/Y106/Y110/Y116-B series antibodies that are expressed at levels greater than 5 mg/L.

Fig. 6 shows the expression level of Y200/Y201/Y203/Y204/Y205/Y206/Y210/Y216-A and B series antibodies having bispecific antibody structure 2 when transiently transfected in CHO cells and the purity determined by size exclusion HPLC after affinity chromatography with protein A.

Fig. 7 shows the expression levels of Y300/Y304/Y316-A series, B series, C series and D series antibodies having bispecific antibody structure 3 when transiently transfected in CHO cells and the purity determined by size exclusion HPLC after Protein A affinity chromatography.

Fig. 8 shows the expression levels of the Y400/Y404/Y416-A series and B series antibodies having bispecific antibody structure 4 when transiently transfected in CHO cells and the purity determined by size exclusion HPLC after Protein A affinity chromatography.

Fig. 9 shows a comparison of the expression level and purity of antibodies with bispecific antibody structure 1-4, with the same target and antibody variable region sequence. (A) Level of expression and purity of antibodies with structure 1-4 bispecific antibodies with the variable region sequence of the antibody S70 (SEQ ID NO: 41 and 42) and with the variable region sequence of the antibody G631 (SEQ ID NO: 57 and 58). (B) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of the antibody S70 (SEQ ID NO: 41 and 42) and with the variable region sequence of the antibody 3G12 (SEQ ID NO: 59 and 60). (C) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of antibody 12A4 (SEQ ID NO: 45 and 46) and with the variable region sequence of antibody 3G12 (SEQ ID NO: 59 and 60). (D) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of the antibody S70 (SEQ ID NO: 41 and 42) and with the variable region sequence of the antibody LAG35 (SEQ ID NO: 33 and 34). (E) Level of expression and purity of antibodies with structure 1, 3 and 4 of the bispecific antibody with the variable region sequence of the antibody 5C4 (SEQ ID NO: 37 and 38) and with the variable region sequence of the Yervoy antibody (SEQ ID NO: 27 and 28). (F) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of the antibody 5C4 (SEQ ID NO: 37 and 38) and with the variable region sequence of the antibody 10A7 (SEQ ID NO: 29 and 30). (G) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of the antibody 5C4 (SEQ ID NO: 37 and 38) and with the variable region sequence of the antibody G631 (SEQ ID NO: 57 and 58). (H) Level of expression and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequence of the antibody 5C4 (SEQ ID NO: 37 and 38) and with the variable region sequence of the antibody B-N10 (SEQ ID NO: 63 and 64). (I) Level of expression and purity of antibodies with structure 1-4 of the bispecific antibody with the variable region sequence of the Elotuzumab antibody (SEQ ID NOs: 49 and 50) and with the variable region sequence of the NM3E2 antibody (SEQ ID NOs: 47 and 48).

Fig. 10 shows the results of an accelerated thermal stability test of various bispecific antibody structures at 40°C. (A) Purity of Y100-B6, Y100-B7, Y200-A1, Y200-B1, Y200-B3, Y300-A1 and Y400-B1 determined by size exclusion HPLC after treatment at 40°C for 0, 7 and 14 days ; (B) Purity of Y101-B1, Y101-B2, Y201-A1, Y201-B1, Y101-B3, Y101-B4, Y201-A2 and Y201-B2 determined by size exclusion HPLC after treatment at 40°C for 0. 7 and 14 days; (C) Purity of Y103-B1, Y103-B2, Y203-A1, Y203-B1, Y104-B1, Y104-B2, Y304-A1 and Y404-B1 determined by size exclusion HPLC after treatment at 40°C for 0. 7 and 14 days; (D) purity of Y105-B1, Y105-B2, Y205-A2, Y205-B2, Y106-B1, Y106-B2, Y206-A2, Y206-B1 and Y206-B2 determined by size exclusion HPLC after treatment at 40°C within 0, 7 and 14 days; (E) Purity of Y110-B1, Y110-B2, Y210-A1, Y210-B1, Y116-B1, Y116-B2, Y216-A1, Y216-B1, Y316-A1, Y416-B1 determined by size exclusion HPLC after treatment at 40°C for 0, 7 and 14 days.

Fig. 11 shows the results of acid resistance testing of various bispecific antibody structures. The pH value of the acid solution used is 3.5. Fig. 11A shows the purity of Y100-B6, Y100-B7, Y200-A1, Y200-B1, Y200-B3, Y300-A1 and Y400-B1 determined by size exclusion HPLC after low pH treatment for 0, 30 and 60 minutes; Fig. 11B shows the purity of Y101-B1, Y101-B2, Y201-A1, Y201-B1, Y101-B3, Y101-B4, Y201-A2 and Y201-B2 determined by size exclusion HPLC after treatment under low pH conditions for 0.30 and 60 minutes; Fig. 11C shows the purity of Y103-B1, Y103-B2, Y203-A1, Y203-B1, Y104-B1, Y104-B2, Y304-A1 and Y404-B1 determined by size exclusion HPLC after treatment under low pH conditions for 0.30 and 60 minutes; Fig. 11D shows the purity of Y105-B1, Y105-B2, Y205-A2, Y205-B2, Y106-B1, Y106-B2, Y206-A2, Y206-B1 and Y206-B2 determined by size exclusion HPLC after treatment under low pH conditions in for 0, 30 and 60 minutes; Fig. 11E shows the purity of Y110-B1, Y110-B2, Y210-A1, Y210-B1, Y116-B1, Y116-B2, Y216-A1, Y216-B1, Y316-A1, Y416-B1 determined by size exclusion HPLC after treatment in low pH conditions for 0, 30 and 60 minutes.

Fig. 12 shows the results of an in vivo pharmacodynamic experiment of Y100-B7 with bispecific antibody structure 1. MC38 mouse colon cancer is used as the tumor cell line, and female C57BL/6 mice are used as the mouse line.

Fig. 13 shows the results of an in vivo pharmacodynamic experiment of Y101-B2 with bispecific antibody structure 1. MC38 mouse colon cancer is used as the tumor cell line, and female C57BL/6 mice are used as the mouse line.

Definitions

It should be noted that a term with the article "a" or "an" refers to one or more of these objects; for example, "bispecific antibody" should be understood as representing one or more bispecific antibodies. Thus, the terms "a" (or "an"), "one or more" and "at least one" may be used interchangeably throughout this application.

As used herein, the term "polypeptide" is intended to cover a single "polypeptide" as well as multiple "polypeptides", and refers to a molecule consisting of monomers (amino acids) linearly linked by amide bonds (also known as peptide bonds). The term "polypeptide" refers to any chain or chains of two or more amino acids, and does not indicate a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, "protein", "amino acid chain" or any other term used referring to a chain or chains of two or more amino acids are all included within the definition of "polypeptide", and the term "polypeptide" may used in place of or interchangeably with any of these terms. The term "polypeptide" is also intended to refer to the products of post-expression modifications of the polypeptide, including, but not limited to, glycosylation, acetylation, phosphorylation, amidation, derivatization with known protecting/blocking groups, proteolytic cleavage, or modification with non-naturally occurring amino acids. The polypeptide may be obtained from a natural biological source or may be produced using recombinant technology, but does not necessarily have to be translated from a specific nucleic acid sequence. It can be obtained by any method, including chemical synthesis.

As used herein, the term “recombinant,” when referring to polypeptides or polynucleotides, refers to a form of the polypeptide or polynucleotide that does not exist in nature, a non-limiting example of which can be obtained by combining polynucleotides or polypeptides that do not normally occur together.

"Homology" or "identity" or "similarity" refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing positions in each sequence, which can be aligned for comparison purposes. If a position in the sequence being compared is occupied by the same base or amino acid, then the molecules are homologous at that position. The degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. An "unrelated" or "non-homologous" sequence has less than 40% identity, although preferably less than 25% identity, with one of the sequences of the present invention.

A polynucleotide or polynucleotide region (or polypeptide or polypeptide region) having a specified percentage (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) of identity sequence" with another sequence means that in an alignment, that percentage of bases (or amino acids) is the same when comparing the two sequences. This alignment and the percentage of homology or sequence identity can be determined using computer programs known in the art, for example the programs described in Ausubel et al. eds. (2007) Current Protocols in Molecular Biology. Preferably, the default settings are used for alignment. One alignment program is BLAST, which uses default parameters. In particular, the BLASTN and BLASTP programs use the following default parameters: Genetic code=standard; filter=no; chain=both; cutoff=60; wait=10; Matrix=BLOSUM62; Descriptions=50 sequences; sort by=HIGH SCORE; Databases=non-redundant, GenBank+EMBL+DDBJ+PDB+GenBank CDS translations+SwissProtein+SPupdate+PIR. Detailed information about these programs can be found at the following Internet address: http://www.ncbi.nlm.nih.gov/blast/Blast.cgi, last accessed May 21, 2008. Biologically equivalent polynucleotides are polynucleotides having the above specified percentage homology and encoding a polypeptide having the same or similar biological activity.

The term "encode" as applied to polynucleotides refers to a polynucleotide that "encodes" a polypeptide and which, in its native state or when manipulated by methods well known to those skilled in the art, can be transcribed and/or translated to produce mRNA for the polypeptide and/or fragment thereof . The antisense strand is the complement of such a nucleic acid, and the coding sequence can be deduced from it.

As used herein, the term “antibody” or “antigen-binding polypeptide” refers to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen. The antibody may be an intact antibody and any antigen binding fragment or one chain thereof. Thus, the term "antibody" includes any protein or peptide containing a specific molecule, where the specific molecule includes at least a portion of an immunoglobulin molecule having antigen binding biological activity. Examples include, but are not limited to, a heavy or light chain complementarity determining region (CDR) or a ligand binding portion thereof, a heavy or light chain variable region, a heavy or light chain constant region, a framework (FR) region, or any portion thereof, or at least one part of the binding protein.

The term "antibody fragment" or "antigen binding fragment" as used herein means a portion of an antibody such as F(ab') ₂ , F(ab) ₂ , Fab', Fab, Fv, Fd, Fv, dAb, Fab/c, complementarity determining region (CDR) fragments linked by disulfide bond Fv (sdFv), single chain antibodies (eg, scFv), divalent antibodies or domain antibodies, and the like. Regardless of structure, the antibody fragment binds to the same antigen that is recognized by the intact antibody. The term "antibody fragment" includes aptamers, spiegelmers and diabodies. The term "antibody fragment" also includes any synthetic or genetically engineered protein that acts like an antibody by binding to a specific antigen to form a complex.

"Single chain variable fragment" or "scFv" refers to a fusion protein of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulins. In some aspects, the regions are connected by a short linker peptide of 10 to about 25 amino acids. The linker can be enriched with glycine for flexibility and serine or threonine for solubility, and can either connect the N-terminus of VH to the C-terminus of VL, or vice versa. This protein retains the specificity of the original immunoglobulin, but the constant region is removed and a linker is introduced. ScFv molecules are known in the art and are described, for example, in US Pat. No. 5,892,019.

The term "antibody" covers a wide range of polypeptides that can be biochemically recognized. Those skilled in the art will appreciate that heavy chains are classified as gamma, mu, alpha, delta, or epsilon (γ, μ, α, δ, ε), with some subclasses among them (eg, γ1-γ4). It is the nature of this chain that determines the "class" of the antibody as IgG, IgM, IgA, IgG or IgE, respectively. Immunoglobulin subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgG5, etc., are well characterized and functionally specific. Modified versions of each of these classes and isotypes are readily recognizable to those skilled in the art given the present disclosure and are accordingly included within the scope of the present disclosure. All classes of immunoglobulins are clearly included within the scope of the present invention; the following discussion will mainly be directed to the IgG class of immunoglobulin molecules. For IgG, a standard immunoglobulin molecule comprises two identical light chain polypeptides with a molecular weight of approximately 23,000 daltons and two identical heavy chain polypeptides with a molecular weight of 53,000-70,000. These four chains are typically linked by disulfide bonds in a "Y" configuration, in which the light chains flank the heavy chains starting at the mouth of the "Y" and extending through the variable region.

Antibodies, antigen binding polypeptides, variants or derivatives thereof in the present invention include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized, primatized or chimeric antibodies, single chain antibodies, antigen binding fragments, such as Fab, Fab' and F(ab' )2, Fd, Fv, single chain Fv (scFv), disulfide-linked single chain Fv (sdFv), fragments including either the VL or VH domain, fragments produced by a Fab expression library, and anti-idiotypic (anti-Id) antibodies. The immunoglobulin or antibody molecules of the present invention may be of any type (eg, IgG, IgE, IgM, IgD, IgA and IgY), any class (eg, IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass of immunoglobulin molecules.

Light chains are classified as kappa or lambda (κ, λ). Each heavy chain class can be linked to either a kappa or lambda light chain. Typically, the light and heavy chains are covalently linked to each other, and the tail portions of the two heavy chains are linked to each other by covalent disulfide bonds or non-covalent bonds when immunoglobulins are generated by hybridomas, B cells or genetically modified host cells. In the heavy chain, the amino acid sequences extend from the N-terminus at the branched ends of the Y-configuration to the C-terminus at the bottom of each chain.

Both light and heavy chains are divided into structural regions and regions of functional homology. The terms "constant" and "variable" are used functionally. In this regard, it should be understood that both the light (VL) and heavy (VH) chain variable domains determine antigen recognition and antigen specificity. Conversely, the light chain (CL) and heavy chain constant domains (CH1, CH2 or CH3) confer important biological properties such as secretion, transplacental motility, Fc receptor binding, complement fixation, etc. Typically, the number of constant region domains increases as they become more distant from the antigen binding site or amino terminus of the antibody. The N-terminal part is the variable region, and the C-terminal part is the constant region; The CH3 and CL domains actually comprise the carboxy terminus of the heavy and light chains, respectively.

As stated above, the variable region allows the antibody to selectively recognize and specifically bind to an epitope on an antigen. That is, the VL domain and VH domain or subclasses of complementarity determining regions (CDRs) of an antibody combine to form a variable region that defines a three-dimensional antigen binding site. This tetravalent antibody structure forms an antigen binding site that is present at the end of each arm in a Y configuration. More specifically, the antigen binding site is defined by three CDRs on each of the VH and VL chains (ie, CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3). In some examples, such as some immunoglobulins derived from camelid species or engineered from camelid immunoglobulins, their intact immunoglobulin molecule may consist of only heavy chains with no light chains. See, for example, Hamers-Casterman et al., Nature 363:446-448 (1993).

In natural antibodies, the six "complementarity determining regions" or "CDRs" present in each antigen binding domain are short, non-contiguous amino acid sequences that are specifically positioned to form the antigen binding domain. The remaining amino acids in the antigen-binding domain, called the “framework” region, exhibit less intramolecular variability. The framework regions generally adopt a β-sheet configuration, and the CDRs form loops that connect the β-sheet structure and sometimes form part of the β-sheet structure. Therefore, framework regions are used to form a scaffold that positions the CDRs in the correct direction through interchain non-covalent interactions. The antigen binding domain formed by the located CDR defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface facilitates non-covalent binding of the antibody to its homologous epitope. Those skilled in the art can easily identify the amino acids of the CDRs and framework regions for any given heavy or light chain variable region since they have been clearly defined (see "Sequences of Proteins of Immunological Interest", Kabat, E., et al., US Department of Health and Human Services, (1983), Chothia and Lesk, J. MoI. Biol., 196:901-917 (1987), the full text of which is incorporated herein by reference).

In the event that there are two or more definitions of a term that are used and/or accepted in the art, the definitions of the term used in this application are intended to include all meanings unless expressly stated otherwise. A specific example is the use of the term “complementarity determining region” (“CDR”) to describe non-contiguous antigen binding sites present in the variable regions of both heavy and light chain polypeptides. This specific area has been described by Kabat et al., US Department of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983) and Chothia et al. , J. MoI. Biol. 196:901-917 (1987), the full text of which is incorporated herein by reference. As defined by Kabat and Chothia, a CDR includes overlapping amino acid residues or amino acid substructures when compared to each other. However, the use of each definition to refer to a CDR antibody or variant thereof will be within the scope of the term as defined and used herein. The corresponding amino acid residues that span the CDRs, as defined in each of the references above, are listed in the table below for comparison. The exact number of residues that span a particular CDR will vary depending on the sequence and the size of the CDR. Those skilled in the art can routinely determine which residues constitute a particular CDR if the amino acid sequence of the antibody variable region is provided.

Table 1
Definition of Antibody Variable Region Kabat Chothia CDR-H1 31-35 26-32 CDR-H2 50-65 52-58 CDR-H3 95-102 95-102 CDR-L1 24-34 26-32 CDR-L2 50-56 50-52 CDR-L3 89-97 91-96

Kabat et al. also defined a variable domain sequence numbering system that is applicable to any type of antibody. Those skilled in the art can clearly use this "Kabat numbering" system for any variable domain sequence, regardless of any experimental data other than the sequence itself. "Kabat numbering" as used herein refers to the numbering system described by Kabat et al., U.S. Department of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983).

In addition to the table above, the CDR regions described by the Kabat numbering system are as follows: CDR-H1 begins at amino acid position 31 (i.e., approximately 9 residues after the first cysteine residue) and includes approximately 5 to 7 amino acids and ends at the next tryptophan residue. CDR-H2 begins at the fifteenth residue after the end of CDR-H1, comprises approximately 16 to 19 amino acids, and ends at the next arginine or lysine residue. CDR-H3 begins approximately at the thirty-third amino acid residue after the end of CDR-H2; includes from 3 to 25 amino acids; and ends with the sequence W-G-X-G, where X represents any amino acid. CDR-L1 begins with a residue at approximately position 24 (i.e., after the cysteine residue); contains approximately 10 to 17 residues; and ends at the next tryptophan residue. CDR-L2 begins at approximately the sixteenth residue after the end of CDR-L1 and includes approximately 7 residues. CDR-L3 begins at approximately the thirty-third residue after the end of CDR-L2 (i.e., after the cysteine residue); contains approximately 7 to 11 residues and ends with the sequence F or WGXG, where X is any amino acid.

The antibodies described herein can be of any animal origin, including birds and mammals. Preferably, the antibodies are human, mouse, donkey, rabbit, goat, guinea pig, camel, llama, horse or chicken. In another embodiment, the variable region may be derived from a condricthoid (eg, shark).

The term "heavy chain constant region" as used herein includes amino acid sequences derived from immunoglobulin heavy chains. A polypeptide comprising a heavy chain constant region includes at least one of the following: a CH1 domain, a hinge domain (e.g., an upper hinge region, a middle hinge region, and/or a lower hinge region), a CH2 domain, a CH3 domain, or a variant or fragment of the above . For example, an antigen binding polypeptide for use in the present invention may include a polypeptide chain comprising a CH1 domain; a polypeptide comprising a CH1 domain, at least part of a hinge domain and a CH2 domain; a polypeptide chain including a CH1 domain and a CH3 domain; a polypeptide chain comprising a CH1 domain, at least a portion of a hinge domain, and a CH3 domain, or a polypeptide chain comprising a CH1 domain, at least a portion of a hinge domain, a CH2 domain, and a CH3 domain. In another embodiment, the polypeptide of the present invention includes a polypeptide chain including a CH3 domain. In addition, the antibodies used in the present invention may lack at least a portion of the CH2 domain (eg, all or part of the CH2 domain). As stated above, those skilled in the art will appreciate that heavy chain constant regions can be modified to differ in amino acid sequence from naturally occurring immunoglobulin molecules.

The heavy chain constant regions of the antibody disclosed herein can be derived from different immunoglobulin molecules. For example, the heavy chain constant region of a polypeptide may include a CH1 domain derived from an IgG1 molecule and a hinge region derived from an IgG3 molecule. In another example, the heavy chain constant region may include a hinge region that is derived in part from an IgG1 molecule and in part from an IgG3 molecule. In another example, the heavy chain portion may include a chimeric hinge that is partly derived from an IgG1 molecule and partly derived from an IgG4 molecule.

The term "light chain constant region" as used herein includes the amino acid sequence derived from the light chain of an antibody. Preferably, the light chain constant region includes at least one of a kappa constant domain and a lambda constant domain.

“Light chain-heavy chain pair” refers to the combination of a light chain and a heavy chain that can form a dimer due to a disulfide bond between the CL domain of the light chain and the CH1 domain of the heavy chain.

As stated previously, the subunit structures and three-dimensional structures of the constant regions of various immunoglobulins are known. As used herein, the term “VH domain” includes the amino-terminal variable domain of an immunoglobulin heavy chain, and the term “CH1 domain” includes the first (preferably amino-terminal) constant region of an immunoglobulin heavy chain. The CH1 domain is adjacent to the VH domain and is the amino terminus of the hinge region of the immunoglobulin heavy chain molecule.

As used herein, the term “CH2 domain” includes a portion of the heavy chain molecule, for example, from residue at approximately position 244 to residue at position 360 of an antibody in accordance with the generally accepted numbering system (residues at positions 244-360, in accordance with the Kabat numbering system; and residues at positions 231-340, according to the EU numbering system; see Kabat et al., U.S. Department of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983). The CH2 domain is unique because it is closely related to pairs with another domain. In contrast, two N-linked branched carbohydrate chains are inserted between the two CH2 domains of the intact natural IgG molecule. The CH3 domain has been documented to extend from the CH2 domain to the C-terminus of the IgG molecule and comprises about 108 residues.

The term "hinge region" as used herein includes the portion of the heavy chain molecule that links the CH1 domain to the CH2 domain. The hinge region comprises about 25 residues and is flexible, allowing the two N-terminal antigen-binding regions to move independently. The hinge regions can be divided into three different domains: the upper hinge domain, the middle hinge domain and the lower hinge domain (Roux et al., J. Immunol 161:4083 (1998)).

As used herein, the term "disulfide bond" includes a covalent bond formed between two sulfur atoms. The amino acid cysteine includes a thiol group that can form a disulfide bond or bridge with a second thiol group. In most natural IgG molecules, the CH1 and CL regions are linked by a disulfide bond, and the two heavy chains are linked by two disulfide bonds at positions corresponding to positions 239 and 242 in the Kabat numbering system (position 226 or 229 in the EU numbering system).

The term "chimeric antibody" as used herein is intended to mean any antibody in which the immunoreactive regions or sites are derived from or derived from the first species and the constant regions (which may be intact, partial, or modified in accordance with the present invention) obtained from the second type. In some embodiments, the target binding regions or sites will be non-human (eg, mouse or primate) and the constant region will be human.

As used herein, "percent humanization" is calculated by determining the number of backbone amino acid differences (i.e., non-CDR differences) between the humanized domain and the germline domain and subtracting that number from the total number of amino acids, which is then divided by the total number of amino acids and multiply by 100.

So-called “specific binding” or “specific to” generally means that the antibody binds to an epitope of an antigen through its antigen binding domain, and that this binding entails some complementarity between the antigen binding domain and the epitope of the antigen. According to this definition, an antibody is said to “specifically bind” to an epitope of an antigen when it binds to an epitope of an antigen, and this binding through the antigen-binding domain is easier than through binding to a random, unrelated epitope of the antigen. The term "specificity" is used herein to define the affinity of a particular antibody to bind to a particular epitope of an antigen. For example, antibody "A" may be said to have higher specificity for a given antigenic epitope than antibody "B", or antibody "A" may be said to have higher specificity for binding to epitope "C" than for binding with the related epitope "D".

The term "treatment" ("treat" or "treating") as used herein refers to both therapeutic treatment and prophylactic or prophylactic measures whose purpose is to prevent or slow down (reduce) an undesirable physiological change or disorder, such as like cancer progression. Beneficial or desired clinical outcomes include, but are not limited to, relief of symptoms, reduction in the severity of the disease, stabilization (eg, prevention of worsening) of the disease state, delay or slowing of disease progression, relief or palliation of the disease, and relief (partial or complete), regardless of whether it is detectable or not. “Treatment” can also mean an increase in survival time compared to expected survival without treatment. Those in need of treatment include those who already have a disease or symptom, and those who are predisposed to the disease or symptom, or those whose disease or symptom needs to be prevented.

Embodiments of the present invention provide various bispecific antibodies that comprise two different or identical antigen-binding polypeptide units. The domain of an antibody that binds an antigen is a Fab, or ScFv, or a non-covalent pairing (Fv) between a heavy chain variable region (VH) and a light chain variable region (VL). In particular, all of these bispecific antibodies have an Fc portion of an antibody heavy chain constant region, and the Fc includes: (1) a hinge, (2) a second heavy chain constant region (CH2), and a third heavy chain constant region (CH3).

Any of the above antibodies or polypeptides may also include additional polypeptides, for example, an encoded polypeptide as described herein, a signal peptide at the N-terminus of the antibody used to direct secretion, or other heterologous polypeptides as described herein.

Those skilled in the art will also appreciate that the antibodies described herein can be modified to differ in amino acid sequence from the natural binding polypeptides from which they are derived. For example, the polypeptide or amino acid sequence derived from the protein may be similar to the original sequence, for example, it may have a certain percentage identity with the original sequence, for example, it may have 60%, 70%, 75%, 80%, 85% identity. , 90%, 95%, 98% or 99% with the original sequence.

In addition, nucleotide or amino acid substitutions, deletions or insertions can be made, leading to conservative substitutions or changes in regions of "essential" amino acids. For example, the polypeptide or amino acid sequence derived from said protein may be identical to the original sequence except for one or more independent amino acid substitutions, insertions or deletions, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20 or more independent amino acid substitutions, insertions or deletions. In some embodiments, the polypeptide or amino acid sequence derived from the protein has 1 to 5, 1 to 10, 1 to 15, or 1 to 20 independent amino acid substitutions, insertions, or deletions from the original sequence.

In other embodiments, the antigen binding polypeptides of the present invention may include conservative amino acid substitutions.

A "conservative amino acid substitution" is the replacement of an amino acid residue with an amino acid residue having a similar side chain. The field defines families of amino acid residues having similar side chains, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine , glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g. alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g. threonine, valine, isoleucine) and aromatic side chains (eg, tyrosine, phenylalanine, tryptophan, histidine). Therefore, nonessential amino acid residues of immunoglobulin polypeptides are preferably replaced with other amino acid residues from the same side chain family. In another embodiment, the amino acid chain may be replaced by a structurally similar amino acid chain that differs in the order and/or side chain family composition.

Non-limiting examples of conservative amino acid substitutions are presented in the table below, where a similarity score of 0 or higher indicates a conservative substitution between two amino acids.

table 2
Non-limiting list of conservative amino acid substitutions C G P S A T D E N Q H K R V M I L F Y W W -8 -7 -6 -2 -6 -5 -7 -7 -4 -5 -3 -3 2 -6 -4 -5 -2 0 0 17 Y 0 -5 -5 -3 -3 -3 -4 -4 -2 -4 0 -4 -5 -2 -2 -1 -1 7 10 F -4 -5 -5 -3 -4 -3 -6 -5 -4 -5 -2 -5 -4 -1 0 1 2 9 L -6 -4 -3 -3 -2 -2 -4 -3 -3 -2 -2 -3 -3 2 4 2 6 I -2 -3 -2 -1 -1 0 -2 -2 -2 -2 -2 -2 -2 4 2 5 M -5 -3 -2 -2 -1 -1 -3 -2 0 -1 -2 0 0 2 6 V -2 -1 -1 -1 0 0 -2 -2 -2 -2 -2 -2 -2 4 R -4 -3 0 0 -2 -1 -1 -1 0 1 2 3 6 K -5 -2 -1 0 -1 0 0 0 1 1 0 5 H -3 -2 0 -1 -1 -1 1 1 2 3 6 Q -5 -1 0 -1 0 -1 2 2 1 4 N -4 0 -1 1 0 0 2 1 2 E -5 0 -1 0 0 0 3 4 D -5 1 -1 0 0 0 4 T -2 0 0 1 1 3 A -2 1 1 1 2 S 0 1 1 1 P -3 -1 6 G -3 5 C 12

In some embodiments, the antibody may be conjugated to therapeutics, prodrugs, peptides, proteins, enzymes, viruses, lipids, biological response modifiers, pharmaceuticals, or PEGs.

The antibody may be associated or combined with a therapeutic agent, which may include detectable labels such as radiolabels, immunomodulators, hormones, enzymes, oligonucleotides, photoactive therapeutic or diagnostic agents, cytotoxic agents, which may be drugs or toxins, ultrasound enhancing agents , non-radioactive tags, combinations thereof and other such components known in the art.

An antibody can be labeled to enable detection by binding it to chemiluminescent compounds. The presence of chemiluminescently labeled antigen-binding polypeptide is then determined by detecting the luminescence generated during the chemical reaction. Examples of particularly useful chemiluminescent labeling compounds are luminol, isoluminol, thermoluminescent acridinium ester, imidazole, acridinium salt and oxalate ester.

It is also possible to label antibodies for detection using fluorescent emitting metals such as 152Eu or other lanthanide labels. These metals can be attached to the antibody using the following metal chelating groups, such as diethylenetriaminepentaacetic acid (DTPA) or ethylenediaminetetraacetic acid (EDTA). Methods for coupling various fragments to antibodies are well known, see, for example, Arnon et al. , “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy,” in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (ed.), pp. 243-56 (Alan R. Liss, Inc. (1985); Hellstrom et al. , "Antibodies For Drug Delivery," in Controlled Drug Delivery (second edition), Robinson et al. (ed. ), Marcel Dekker, Inc., pp. 623-53 (1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. ( ed.), pp. 475-506 (1985); "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (ed.) , Academic Press pp. 303-16 (1985), and Thorpe et al. , "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev. (52:119-58 (1982))

Method for obtaining antibodies

Methods for producing antibodies are well known in the art and are described herein. In some embodiments, both the variable and constant regions of the antigen binding polypeptides of the present invention are entirely human. Fully human antibodies can be obtained using methods described in the prior art and as described in this application. For example, fully human antibodies against a particular antigen can be obtained by administering the antigen to a transgenic animal that has been modified to produce such antibody in response to antigen stimulation, but whose endogenous loci have been disabled. Examples of methods that can be used to obtain such antibodies are described in US patent Nos.: 6150584; 6458592; 6420140, the entire contents of which are incorporated herein by reference.

The binding specificity of the antigen binding polypeptide of the present invention can be measured by performing in vitro experiments such as immunoprecipitation, radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA).

Alternatively, the method described for preparing single-chain structures (US Pat. No. 4,694,778; Bird, Science 242:423-442 (1988); Huston et al. Proc. Natl. Acad. Sci. USA 55:5879-5883 (1988); and Ward et al. , Nature 334:544-554 (1989)) can be used to prepare the single chain structures of the present invention. Single-chain structures are obtained by linking heavy and light chain fragments of the Fv region through an amino acid bridge, resulting in a single-chain fusion peptide. Methods for synthesizing functional Fv fragments in E. coli (Skerra et al. , Science 242: 1038-1041 (1988)) can also be used.

Examples of methods that can be used to produce single chain Fvs (scFvs) and antibodies include those described in US Pat. Nos. 4,946,778 and 5,258,498; Huston et al. , Methods in Enzymology 203:46-88 (1991); Shu et al. ,Proc. Natl. Sci. USA 90:1995-1999 (1993); and Skerra et al. , Science 240:1038–1040 (1988). For some applications, including in vivo use of the antibody in humans and in vitro detection assays, the use of chimeric, humanized or human antibodies may be preferred. Chimeric antibodies are molecules in which different parts of the antibody are derived from different animal species, such as antibodies containing a variable region from a murine monoclonal antibody and a constant region from a human immunoglobulin. Methods for producing chimeric antibodies are known in the art. See, for example, Morrison, Science 229:1202 (1985); Oi et al. , BioTechniques 4:214 (1986); Gillies et al. , J. Immunol. Methods 125:191-202 (1989); US Patent Nos. 5,807,715; 4816567 and 4816397, the entire contents of which are incorporated herein by reference.

A humanized antibody is an antibody molecule that is derived from a non-human species and binds to a target antigen, and such antibody molecule contains one or more complementarity determining regions (CDRs) from the non-human species and a framework region from a human immunoglobulin molecule. Typically, framework residues in the human framework region will be modified by replacement with corresponding residues from the CDR of the donor antibody, preferably to increase antigen-binding capacity. These framework substitutions are identified by methods known in the art, for example, by modeling interactions between CDRs and framework residues to identify framework residues that are important for antigen binding and sequence comparisons to identify abnormal framework residues at specific positions. (See, for example, US Pat. No. 5,585,089 to Queen et al.; Nature 332:323 (1988) to Riechmann et al. , the entire contents of which are incorporated herein by reference). Antibodies can be humanized using various methods known in the art, including, for example, CDR grafting (EP 239400; PCT Publication No. WO 91/09967; US Patent Nos. 5225539; 5530101 and 5585089), masking of surface residues, or alteration surface variable domains (EP 592106; EP 519596; Padlan, Molecular Immunology 28(4/5):489-498 (1991); Studnicka et al. , Protein Engineering 7(6):805-814 (1994); Roguska et al. . , Proc. Natl. Sci. USA 91:969-973 (1994)) and strand permutation (US Pat. No. 5,565,332, the entire contents of which are incorporated herein by reference).

Using conventional recombinant DNA technology, one or more CDRs of the antigen binding polypeptide of the present invention can be inserted into a framework region, for example inserted into a human framework region to humanize non-human antibodies. The framework region can be a natural or heterologous framework region and is preferably a human framework region (see, for example, Chothia et al., J. Mol. Biol. 278:457-479 (1998), for a list of human framework regions). Preferably, the polynucleotide obtained by the combination of the framework region and the CDR encodes a polypeptide that specifically binds to at least one antigenic epitope of the desired polypeptide, for example, LIGHT. Preferably, one or more amino acid substitutions can be made in the framework region, and preferably the amino acid substitutions improve binding of the antibody to its antigen. In addition, this method can be used for amino acid substitutions or deletions of one or more variable region cysteine residues (cysteine residues are involved in the formation of intrachain disulfide bonds), thereby creating antibody molecules lacking one or more intrachain disulfide bonds. Other changes made to polynucleotides are included within the scope of the present invention and the scope of the prior art.

In addition, methods for producing “chimeric antibodies” by splicing genes from mouse antibody molecules can be used (Morrison et al., Proc. Natl. Acad. Sci. USA: 851-855 (1984); Neuberger et al., Nature 37:604 -608 (1984); Takeda et al. Nature 314:452-454 (1985)), and a molecule with the appropriate antigen specificity will be linked to the gene of a human antibody molecule with the appropriate biological activity. As used herein, the term “chimeric antibody” means a molecule in which different parts are derived from different animal species, for example, antibodies containing a murine monoclonal antibody variable region and a human immunoglobulin constant region.

However, another effective method for producing recombinant antibodies is disclosed in Newman, Biotechnology 10: 1455-1460 (1992). In particular, this technique led to the production of primatized antibodies containing monkey variable domains and human constant sequences. This document is incorporated herein by reference in its entirety. In addition, this method is also described in co-assigned U.S. Patent Nos. 5,658,570, 5,693,780, and 5,756,096, each of which is incorporated herein by reference.

Alternatively, antibody-producing cell lines can be selected and cultured using methods well known to those skilled in the art. Such methods are described in various laboratory manuals and primary publications. In this regard, methods suitable for use in the present invention are described, for example, in Current Protocols in Immunology, edited by Coligan et al ., Green Publishing Associates and Wiley-Interscience, John Wiley and Sons, New York (1991), which are incorporated by links in their entirety, including additional links.

In addition, standard methods known to those skilled in the art can be used to introduce mutations into the nucleotide sequences encoding the antibodies of the present invention, including, but not limited to, site-directed mutagenesis and PCR-mediated mutations that result in amino acid substitutions. Preferably, variants (including derivatives) compared to the reference heavy chain variable region, CDR-H1, CDR-H2, CDR-H3, light chain variable region, CDR-L1, CDR-L2 or CDR-L3, encode fewer than 50 amino acid substitutions, less than 40 amino acid substitutions, less than 30 amino acid substitutions, less than 25 amino acid substitutions, less than 20 amino acid substitutions, less than 15 amino acid substitutions and less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions or less than 2 amino acid substitutions. Alternatively, mutations can be introduced randomly along all or part of the coding sequence, for example, by saturation mutagenesis, and the resulting mutants can be screened for biological activity to identify mutations that retain activity.

Antibody structure information

Monospecific antibodies are symmetrical antibodies containing two identical light chains and two identical heavy chains. The light chain and heavy chain are linked by a disulfide bond and target the corresponding antigen; the heavy chain and the heavy chain are linked by a disulfide bond; the antibody as a whole has a "Y" configuration. The light chain includes a light chain variable region (VL) and a light chain constant region (Lc), and the heavy chain includes a heavy chain variable region (VH) and a heavy chain constant region, where the heavy chain constant region includes CH1 and Fc, and Fc includes a hinge , CH2 and CH3.

Structure 1 of the bispecific antibody is F(ab) ₂ -(Fv) ₂ -Fc, which is a tetravalent symmetric bispecific antibody comprising two identical hybrid heavy chains and two identical hybrid light chains, where the two hybrid heavy chains form a pair, the hybrid light chain and the hybrid heavy chain form a pair, and each pair forms one or more interchain disulfide bonds. The hybrid antibody heavy chain includes an antibody heavy chain variable region a (VHa), a first constant region (CH1), an antibody b variable region 1, and an Fc fragment. Antibody b variable region 1 is located between CH1 and Fc and is linked by a linker. The hybrid antibody light chain includes an antibody light chain variable region a (VLa), a light chain constant region (CL) and an antibody b variable region 2. Antibody b variable region 2 is located at the C-terminus of CL and is linked by a linker; where (1) antibody b variable region 1 is a heavy chain variable region (VHb), antibody b variable region 2 is a light chain variable region (VLb), and the antibody is called a structure 1 A series bispecific antibody, or (2) variable region Antibody b 1 is a light chain variable region (VLb), Antibody b variable region 2 is a heavy chain variable region (VHb), and the antibody is called a B series bispecific antibody of structure 1. Where the VHa-VLa pair targets antigen A, and VHb -VLb pair targets the B antigen.

Fig. 1A schematically represents the structure of the bispecific antibody of Structure 1, and FIG. 1B schematically represents the primary protein structure of each antibody component.

Bispecific antibody structure 2 is IgG(H)-ScFv, which is a tetravalent symmetric bispecific antibody comprising two identical hybrid heavy chains and two identical light chains, where the two hybrid heavy chains form a pair, the light chain and the hybrid heavy chain form a pair, and each pair forms one or more interchain disulfide bonds; the hybrid heavy chain includes a heavy chain variable region (VHa), a first heavy chain constant region (CH1), an Fc, and a ScFv, wherein the ScFv is located at the C-terminus of the Fc and is linked by a linker. The light chain-heavy chain pair targets the A antigen, and ScFv targets the B antigen.

Fig. 2A schematically represents the structure of a bispecific antibody of structure 2, and FIG. 2B schematically represents the primary protein structure of each antibody component.

Bispecific antibody structure 3 is Tandem-ScFv-Fc, which is a tetravalent symmetric bispecific antibody comprising two identical hybrid peptides. The fusion peptide includes ScFv (ScFv-a) targeting antigen A, (ScFv-b) targeting antigen B, and Fc, where ScFv-b is located between ScFv-a and Fc and is connected by a linker or hinge.

Fig. 3A schematically represents the structure of a bispecific antibody of structure 3, and FIG. 3B schematically represents the primary protein structure of each antibody component.

Bispecific antibody structure 4 is DVD-IgG, which is a tetravalent symmetric bispecific antibody including two identical hybrid light chains and two identical hybrid heavy chains, where two hybrid heavy chains form a pair, and two hybrid light chains and hybrid heavy chains form a pair , and each pair forms one or more interchain disulfide bonds; the hybrid light chain includes antibody b variable region 1 and antibody a light chain, which is linked by a linker wherein the antibody a light chain is located at the C-terminus; the hybrid heavy chain includes antibody b variable region 2 and antibody a heavy chain, which is linked by a linker where the antibody a heavy chain is located at the C-terminus. Where (1) antibody b variable region 1 is a light chain variable region (VLb), antibody b variable region 2 is a heavy chain variable region (VHb), or (2) antibody b variable region 1 is a heavy chain variable region ( VHb), antibody b variable region 2 is a light chain variable region (VLb). The VHa-VLa pair targets the A antigen, and the VHb-VLb pair targets the B antigen.

Fig. 4A schematically represents the structure of the bispecific antibody of structure 4, and FIG. 4B schematically represents the primary protein structure of each antibody component.

All of the above four bispecific antibody structures are symmetrical structures with an Fc moiety, and all are tetravalent, with two targeting the A antigen and the other two targeting the B antigen.

Variable regions of antibody sequences

Table 3
Anti-CD3 antibody variable region sequences Antibody code
(sequence source) Anti-CD3 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 2a5 QVQLVESGGGVVQPGRSLRLSCAASGTFFS TYAMN
WVRQAPGKGLEWVA RIRSKYNNYATYYADSVKD
RFTISRDDSKNTLYLQMNSLRAEDTAVYYCAR HGN
FGNSYVSWFAY WGQGTLVTVSS 1 QTVVTQEPSLTVSPGGTVTLTC RSSTGAVTT
SNYAN WVQQKPGQAPRGLIG GTNKRAP GV
PARFSGSLLGGKAALTLSGVQPEDEAEYYC A
LWYSNLWV FGGGTKVEIK 2 2j5a QVQLVESGGGVVQPGRSLRLSCAASGTFFS TYAMN
WVRQAPGKGLEWVA RIRSKYNNYATYYADSVKD
RFTISRDDSKNTLYLQMNSLRAEDTAVYYCAR HGN
FGNSYVSWAAY WGQGTLVTVSS 3 QTVVTQEPSLTVSPGGTVTLTC RSSTGAVTT
SNYAN WFQQKPGQAPRGLIG GTNKRAP GVP
ARFSGSLLGGKAALTLSGVQPEDEAEYYC AL
WYSNLWV FGGGTKVEIK 4 I2C
(US8236308) EVQLVESGGGLVQPGGSLKLSCAASGFTFN KYAMN
WVRQAPGKGLEWVA RIRSKYNNYATYYADSVKD R
FTISRDDSKNTAYLQMNNLKTEDTAVYYCVR HGNF
GNSYISYWAY WGQGTLVTVSS 5 QTVVTQEPSLTVSPGGTVTLTC GSSTGAVTS
GNYPN WVQQKPGQAPRGLIG GTKFLAP GTP
ARFSGSLLGGKAALTLSGVQPEDEAEYYC VL
WYSNRWV FGGGTKLTVL 6

Table 4
Anti-B7-H3 antibody variable region sequences Antibody code
(sequence source) Anti-CD38 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 8H9
(Cancer Research 61, 4048-4054, May 15, 2001) QVQLQQSGAELVKPGASVKLSCKASGYTFT NYDIN
WVRQRPEQGLE WIGWIFPGDGSTQYNEKFKG KA
TLTTDTSSSTAYMQLSRLTSEDSAVYFCAR QTTATW
FAY WGQGTLVTVSS 7 DIVMTQSPATLSVTPGDRVSLSC RASQSISDY
LH WYQQKSHESPRLLIK YASQSIS GIPSRFSG
SGSGSDFTLSINSVEPEDVGVYYC QNGHSFP
LT FGAGTKLELK 8 BRCA69D
(US20120294796A1) QVQLQQSGAELARPGASVKLSCKASGYTFT SYWM
Q WVKQRPGQGLEWIG TIYPGDGDTRYTQKFKG K
ATLTADKSSSTAYMQLSSLASEDSAVYYCAR RGIPR
LWYFDV WGAGTTVTVSS 9 DIQMTQTTSSLSSASLGDRVTISC RASQDISNY
LN WYQQKPDGTVKLLIY YTSRLHS GVPSRF
SGSGSGTDYSLTIDNLEQEDIATYFC QQGNTL
PPT FGGGTKLEIK 10

Table 5
Anti-CD38 antibody variable region sequences Antibody code
(sequence source) Anti-CD38 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. Dara
(US9040050) EVQLLESGGGLVQPGGSLRLSCAVSGFTF NSFAMS
WVRQAPGKGLEWVS AISGSGGGTYYADSVKG RF
TISRDNSKNTLYLQMNSLRAEDTAVYFCAK DKILW
FGEPVFDY WGQGTLVTVSS eleven EIVLTQSPATLSLSPGERATLSC RASQSVSSY
LAW YQQKPGQAPRLLIY DASNRA TGIPARFS
GSGSGTDFTLTISSLEPEDFAVYYC QQRSNW
PPT FGQGTKVEIK 12 MOR
(US8088896) QVQLVESGGGLVQPGGSLRLSCAAS GFTFSSYYM
N WVRQAPGKGLEWVS GISGDPSNTYYADSVKG RF
TISRDNSKNTLYLQMNSLRAEDTAVYYCAR DLPLV
YTGFAY WGQGTLVTVSS 13 DIELTQPPSVSVAPGQTARISC SGDNLRHYY
VYW YQQKPGQAPVLVIY GDSKRPS GIPERFS
GSNSGNTATLTISGTQAEDEADYYC QTYTG
GASLV FGGGTKLTVLGQ 14 2F5
(US9040050) QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAFS
WVRQAPGQGLEWMG RVIPFLGIANSAQKFQG RV
TITADKSTSTAYMDLSSLRSEDTAVYYCAR DDIAAL
GPFDY WGQGTLVTVSS 15 DIQMTQSPSSLSSASVGDRVTITC RASQGISSW
LA WYQQKPEKAPKSLIY AASSLQS GVPSRFS
GSGSGTDFTLTISSLQPEDFATYYC QQYNSYP
RT FGQGTKVEIK 16

Table 6
Anti-EpCAM antibody variable region sequences Antibody code
(sequence source) Amino acid sequences of the anti-BCMA antibody variable region (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 3-17I (US20100310463) QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAIS
WVRQAPGQGLEWMG GIIPIFGTANYAQKFQG RV
TITADESTSTAYMELSSLRSEDTAVYYCAR GLLWN
Y WGQGTLVTVSS 17 EIVMTQSPATLSVSPGERATLSC RASQSVSSN
LA WYQQKPGQAPRLIIY GASTTAS GIPARFS
ASGSGTDFTLTISSLQSEDFAVYYC QQYNNW
PPAYT FGQGTKLEIK 18 2-6 (TW102107344) EVQLVESGPELKKPGETVKISCKAS GYTFTDYSMH
WVKQAPGKGLKWMGW INTETGEP TYADDFKGRF
AFSLETSASTAYLQINNLKNEDTATYFCAR TAVY W
GQGTTVTVSS 19 DIQMTQSPSSLSSASLGERVSLTC RASQEISVS
LS WLQQEPDGTIKRLIY ATSTLDS GVPKRFS
GSRSGSDYSLTISSLESEDFVDYYC LQYASYP
WT FGGGTKLEIK 20

Table 7
Anti-BCMA Antibody Variable Region Sequences Antibody code
(sequence source) Amino acid sequences of the anti-BCMA antibody variable region (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. B50
(US9598500) QVQLVQSGAEVKKPGASVKVSCKASGYSFP DYYIN
WVRQAPGQGLEWMG WIYFASGNSEYNQKFTG RV
TMTRDTSINTAYMELSSLTSEDTAVYFCAS LYDYD
WYFDV WGQGTMVTVSS 21 DIVMTQTPLSLSVTPGQPASISC KSSQSLVHS
NGNTYLH WYLQKPGQSPQLLIY KVSNRFS G
VPDRFSGSGSGTDFTLKISRVEAEDVGIYYC S
QSSIYPWT FGQGTKLEIK 22 B140153
(WO2016090320A1) QVQLVQSGAEVKKPGSSVKVSCKAS GGTFSSYA IS
WVRQAPGQGLEWMGR IIPILGIA NYAQKFQGRVTI
TADKSTSTAYMELSSLRSEDTAVYYC ARGGYYSH
DMWSED WGQGTLVTVSS 23 LPVLTQPPSASGTPGQRVTISCSGR SSNIGSNS
VNWYRQLPGAAPKLLIY SNN QRPPGVPVRFS
GSKSGTSASLAISGLQSEDEATYYC ATWDDN
LNVHYV FGTGTKVTVLG 24 B69
(US2017051068A1) QLQLQESGPGLVKPSETLSLTCTVSGGSIS SGSYFW
G WIRQPPGKGLEWIG SIYYSGITYYNPSLKS RVTIS
VDTSKNQFSLKLSSVTAADTAVYYCAR HDGAVAG
LFDY WGQGTLVTVSS 25 SYVLTQPPSVSVAPGQTARITC GGNNIGKS
VH WYQQPPGQAPVVVVY DDSDRPS GIPERF
SGNSNGNTATLTISRVEAGDEAVYYC QVWD
SSSDHVV FGGGTKLTVL 26

Table 8
Anti-CTLA-4 antibody variable region sequences Antibody code
(sequence source) Anti-CTLA-4 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. Yervoy(US20020086014A1) QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH
WVRQAPGKGLEWVTFI SYDGNNKYYADSVKG RFT
ISRDNSKNTLYLQMNSLRAEDTAIYYCAR TGWLGP
FDY WGQGTLVTVSS 27 EIVLTQSPGTLSLSPGERATLSC RASQSVGSS
YLA WYQQKPGQAPRLLIY GAFSRAT GIPDRF
SSGSGSGTDFTLTISRLEPEDFAVYYC QQYGSS
PWT FGQGTKVEIK 28

Table 9
Anti-TIGIT antibody variable region sequences Antibody code
(sequence source) Anti-TIGIT antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 10A7 (US20090258013A1) EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH
WVRQSPGKGLEWVAFI RSGSGIVFYADAVRG RFTI
SRDNAKNLLFLQMNDLKSEDTAMYYCAR RPLGHN
TFDS WGQGTLVTVSS 29 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYY
SGVKENLLA WYQQKPGQSPKLLIY YASIRF
T GVPDRFTGSGSGTDYTLITTSVQAEDMGQY
FC QQGINNPLT FGDGTKLEIK thirty MAB10 (WO2017059095A1) QVQLQESGPGLVKPSQTLSLTCTVSGG SIESGLYY
WG WIRQPPGKGLEWIGSI YYSGSTYYNPSLKS RATI
SVDTSKNQFSLKLSSVTAADTAVYYCAR DGVLAL
NKRSFDI WGQGTMVTVSS 31 EIVLTQSPGTLSLSPGERATLSC RASQSVSSS
YLA WYQQKPGQAPRLLIY GASSRAT GIPDRF
SSGSGSGTDFTLTISRLEPEDFAVYYC QQHTV
RPPLT FGGGTKVEIK 32

Table 10
Anti-LAG-3 antibody variable region sequences Antibody code
(sequence source) Anti-LAG-3 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. LAG35 (US9505839B2) QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYW
N WIRQPPGKGLEWIGEI NHRGSTNSNPSLKS RVTLS
LDTSKNQFSLKLRSVTAADTAVYYCA FGYSDYEY
NWFDP WGQGTLVTVSS 33 EIVLTQSPATLSLSPGERATLSC RASQSISSYL
A WYQQKPGQAPRLLIY DASNRAT GIPARFSG
SGSGTDFTLTISSLEPEDFAVYYC QQRSNWP
LT FGQGTNLEIK 34 L3E3(US9902772B2) EVQLLESGAEVKKPGASVKVSCKASGYTFT SYYM
H WVRQAPGQGLEWMGI INPSAGSTSYAQKFQG RV
TMTRDTSTSTVYMELSSLRSEDTAVYYCAR ELMAT
GGFDY WGQGTLVTVSS 35 QSVLTQPASASGSPGQSITISC TGTSSDVGGY
NYVS WYQQHPGKAPKL MIYDVSNRPS GVS
NRFSGSKSGNTASLTISGLQAEDEANYYC SS
YTSSSTNV FGTGTKVTVL 36

Table 11
Anti-PD-1 antibody variable region sequences Antibody code
(sequence source) Anti-PD-1 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 5C4 (WO2006121168) QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH
WVRQAPGKGLEWVAVI WYDGSKRYYADSVKG RF
TISRDNSKNTLFLQMNSLRAEDTAVYYCA TNDDY
WGQGTLVTVSS 37 EIVLTQSPATLSLSPGERATLSC RASQSVSSY
LA WYQQKPGQAPRLLIY DASNRAT GIPARFS
GSGSGTDFTLTISSLEPEDFAVYYC QQSSNW
PRT FGQGTKVEIK 38 H409A11 (WO2008156712A1) QVQLVQSGVEVKKPGASVKVSCKASGYTFT NYYM
Y WVRQAPGQGLEWMGG INPSNGGTNFNEKFKN R
VTLTTDSSTTTAYMELKSLQFDDTAVYYCAR RDYR
FDMGFDY WGQGTTVTVSS 39 EIVLTQSPATLSLSPGERATLSC RASKGVSTS
GYSYLH WYQQKPGQAPRLLIY LASYLES GV
PARFSGSGSGTDFTLTISSLEPEDFAVYYC QH
SRDLPLT FGGGTKVEIK 40

Table 12
Anti-PD-L1 antibody variable region sequences Antibody code
(sequence source) Anti-PD-L1 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. S70
(WO2010077634A1) EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH
WVRQAPGKGLEWVAWI SPYGGSTYYADSVKG RFT
ISADTSKNTAYLQMNSLRAEDTAVYYCAR RHWPG
GFDY WGQGTLVTVSS 41 DIQMTQSPSSLSSASVGDRVTITC RASQDVST
AVA WYQQKPGKAPKLLIY SASFLYS GVPSRF
SSGSGSGTDFTLTISSLQPEDFATYYC QQYLY
HPAT FGQGTKVEIK 42 Avelumab
(WO2013079174A1) EVQLLESGGGLVQPGGSLRLSCAASGFTF SSYIMM
WVRQAPGKGLEWVS SIYPSGGITFYADTVKG RFTI
SRDNSKNTLYLQMNSLRAEDTAVYYCAR IKLGTVT
TVDY WGQGTLVTVSS 43 QSALTQPASVSGSPGQSITISC TGTSSDVGGY
NYVS WYQQHPGKAPKLMIY DVSNRPS GVSN
RFSGSKSGNTASLTISGLQAEDEADYYC SSYT
SSSTRV FGTGTKVTVL 44 12A4(US7943743B2) QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS
WVRQAPGQGLEWMGGI IPIFGKAHYAQKFQG RV
TITADESTSTAYMELSSLRSEDTAVYFCAR KFHFVS
GSPFGMDV WGQGTTVTVSS 45 EIVLTQSPATLSLSPGERATLSC RASQSVSSY
LA WYQQKPGQAPRLLIY DASNRAT GIPARFS
GSGSGTDFTLTISSLEPEDFAVYYC QQRSNW
PT FGQGTKVEIK 46

Table 13
Anti-CD16 antibody variable region sequences Antibody code
(sequence source) Anti-CD16 antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS
WVRQAPGKGLEWVSG INWNGGSTGYADSVKG RF
TISRDNAKNSLYLQMNSLRAEDTAVYYCAR GRSLL
FDY WGQGTLVTVSR 47 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS
WYQQKPGQAPVLVIY GKNNRPS GIPDRFSGS
SSGNTASLTITGAQAEDEADYYC NSRDSSGN
HVV FGGGTKLTVL 48

Table 14
Anti-SLAMF7 antibody variable region sequence Antibody code
(sequence source) Amino acid sequence of the anti-SLAMF7 antibody variable region (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. Elotuzumab
(WO2004100898A2) EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS
WVRQAPGKGLEWIGE INPDSST I NYAPSLKDKFIISR
DNAKNSLYLQMNSLRAEDTAVYYC ARPDGNYWY
FDV WGQGTLVTVSS 49 DIQMTQSPSSLSSASVGDRVTITCKAS QDVGI
A VAWYQQKPGKVPKLLIY WAS TRHTGVPDR
FSGSGSGTDFTLTISSLQPEDVATYYC QQYSS
YPYT FGQGTKVEIK 50

Table 15
Anti-CEA antibody variable region sequences Antibody code
(sequence source) Amino acid sequences of the anti-CEA antibody variable region (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. hPR1A3(Cancer Immunol Immunother (1999) 47:299-306) QVQLVQSGSELKKPGASVKVSCKASGYTFT VFGM
N WVRQAPGQGLEWMG WINTKTGEATYVEEFKG
RFVFSLDTSVSTAYLQISSLKADDTAVYYCAR WDF
YDYVEAMDY WGQGTTVTVSS 51 DIQMTQSPSSLSSASVGDRVTTITC KASQNVGT
NVA WYQQKPGKAPKLLIY SASYRYS GVPSR
FSGSGSGTDFTFTISSLQPEDIATYYC HQYYT
YPLFT FGQGTKVEIK 52

Table 16
Anti-VEGF antibody variable region sequences Antibody code
(sequence source) Anti-VEGF antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. Avastin EVQLVESGGGLVQPGGSLRLSCAAS GYTFTNYGM
N WVRQAPGKGLEWVG WINTYTGEPTYAADFKR R
FTFSLDTSKSTAYLQMNSLRAEDTAVYYCAK YPHY
YGSSHWYFDV WGQGTLVTVSS 53 DIQMTQSPSSLSSASVGDRVTITTC SASQDISNY
LN WYQQKPGKAPKVLIY FTSSLHS GVPSRFS
GSGSGTDFTLTISSLQPEDFATYYC QQYSTVP
WT FGQGTKVEIK 54 B2041 (WO2005012359A2) EVQLVESGGGLVQPGGSLRLSCAASGFSIN GSWIF
WVRQAPGKGLEWV GAIWPFGGYTH YADSVKGRF
TISADTSKNTAYLQMNSLRAEDTAVYYCAR WGHS
TSPWAMDY WGQGTLVTVSS 55 DIQMTQSPSSLSSASVGDRVTITC RASQVIRRS
LA WYQQKPGKAPKLLIY AASNLAS GVPSRFS
GSGSGTDFTLTISSLQPEDFATYYC QQSNTSP
LT FGQGTKVEIK 56 G631 (WO2005012359A2) EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH
WVRQAPGKGLEWVA GITPAGGYTYYADSVKG RF
TISADTSKNTAYLQMNSLRAEDTAVYYCAR FVFFL
PYAMDY WGQGTLVTVSS 57 DIQMTQSPSSLSSASVGDRVTITC RASQDVST
AVA WYQQKPGKAPKLLIY SASFLYS GVPSRF
SSGSGSGTDFTLTISSLQPEDFATYYC QQGYG
NPFT FGQGTKVEIK 58

Table 17
Anti-TGF-β antibody variable region sequences Antibody code
(sequence source) Anti-TGF-β antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS
WVRQAPGQGLEWMG GVIPIVDIANYAQ RFKGRVT
ITADESTSTTYMELSSLRSEDTAVYYCA STLGLVLD
AMDY WGQGTLVTVSS 59 ETVLTQSPGTLSLSPGERATLSC RASQSLGSS
YLA WYQQKPGQAPRLLIY GASSRAP GIPDRF
SSGSGSGTDFTLTISRLEPEDFAVYYC QQYAD
SPIT FGQGTRLEIK 60 4B9 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS
WVRQAPGQGLEWMG GVIPIVDIANYAQ RFKGRVT
ITADESTSTTYMELSSLRSEDTAVYYCA LPRAFVLD
AMDY WGQGTLVTVSS 61 ETVLTQSPGTLSLSPGERATLSC RASQSLGSS
YLA WYQQKPGQAPRLLIY GASSRAP GIPDRF
SSGSGSGTDFTLTISRLEPEDFAVYYC QQYAD
SPIT FGQGTRLEIK 62

Table 18
Anti-IL-10 antibody variable region sequences Antibody code
(sequence source) Anti-TGF-β antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH
WVRQSPGKGLEWLGVIWRG GSTDYSAAFMS RLSI
TKDNSKSQVFFKMNSLQADDTAIYFCAK QAYGHY
MDY WGQGTSVTVSS 63 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHS
NGNTYLE WYLQKPGQSPKLLIY KVSNRFS G
VPDRFSGSGSGTDFTLKITRLEAEDLGVYYC
FQGSHVPWT FGGGTKLEIK 64 BT-063 EVQLVESGGGLVQPGGSLRLSCAAS GFSFATYGVH
WVRQSPGKGLEWLGVIWRG GSTDYSAAFMS RLTI
SKDNSKNTVYLQMNSLRAEDTAVYFCAK QAYGH
YMDY WGQGTSVTVSS 65 DVVMTQSPLSLPVTLGQPASISC RSSQNIVHS
NGNTYLE WYLQRPGQSPRLLIY KVSNRFS G
VPDRFSGSGSGTDFTLKISRVEAEDVGVYYC
FQGSHVPWT FGQGTKVEIK 66

Table 19
Anti-luciferase antibody variable region sequences Antibody code
(sequence source) Anti-luciferase antibody variable region amino acid sequences (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. 4420 evkldetggglvqpgrpmklscvasgftfs dywm
n wvrqspekglewva qirnkpynyetyysdsvk
g rftisrddskssvylqmnnlrvedmgiyyctg sy
ygmdy wgqgtsvtvss 67 dvvmtqtplslpvslgdqasisc rssqslvh
sngntylr wylqkpgqspkvliy kvsnrfs
gvpdrfsgsgsgtdftlkisrveaedlgvyf
c sqsthvpwt fgggtkleik 68

Sequence of other domains

(1) Amino acid sequences of the linker domain

Table 20
Linker amino acid sequences Domain Code Amino acid sequences SEQ ID NO. Linkers Lin1 SS 69 Lin2 AS 70 Lin3 GGGGS 71 Lin4 GGGSAAA 72 Lin5 GGGGSAS 73 Lin6 GYPGGGGS 74 Lin7 GGGGSGGGGS 75 Lin8 APAPAPAPAPAP 76 Lin9 AEAAAAKEAAAKA 77 Lin10 GKSSGSGSESKS 78 Lin11 GSTSGSGKSSEGKG 79 Lin12 EPKSSDKTHTSPPS 80 Lin13 GGGGSDKTHTSPPS 81 Lin14 GGGGSGGGGSGGGGS 82 Lin15 GAAAGGGGSGGGGS 83 Lin16 EAAAKGGGGSGGGGS 84 Lin17 GGGGSEAAAKGGGGS 85 Lin18 EAAAAKEAAAKGGGGS 86 Lin19 AGGGGSGGGGSGGGGSG 87 Lin20 GSTSGSGKSSEGSGSTKG 88 Lin21 GAAAGGGGSGGGGSGGGGS 89 Lin22 GGGGSGGGGSGGGGSGGGGS 90

(2) Amino acid sequences of the hinge domain

Table 21
Amino acid sequences of the hinge Domain Code Amino acid sequences SEQ ID NO. Hinges Hin1 DKTHTCP 91 Hin2 ERKCCVECP 92 Hin3 ELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPR 93 Hin4 ESKYGPPCPSCP 94 Hin5 DKTHT 95 Hin6 ESKYGPPCPPCP 96 Hin7 EPKSSDKTHTCP 97 Hin8 GGGGSDKTHTCP 98 Hin9 RGRGSDKTHTCP 99 Hin10 DGDGSDKTHTCP 100 Hin11 GRGRGSDKTHTCP 101 Hin12 ASTRGRGSDKTHTCP 102 Hin13 GQPDGDASDKTHTCP 103

(3) Amino acid sequence of the CL domain of the light chain constant region

Table 22
Amino acid sequences CL Domain Code Amino acid sequences SEQ
ID NO. C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 104 Lc2 GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS 105 Lc3 GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS 106 Lc4 GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPAKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS 107 Lc5 GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVKVAWKADGSPVNTGVETTTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPAECS 108 Lc6 GQPKAAPTVTLFPPSSEELQANKATLVCLISDFYPGAVKVAWKADSSPAKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHKSYSCQVTHEGSTVEKTVAPTECS 109 Lc7 VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 110

(4) Amino acid sequences of the CH1 domain of the heavy chain constant region

Table 23
Amino acid sequences of CH1 Domain Code Amino acid sequences SEQ
ID NO. CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC 111 G2CH1 ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTV 112 G3CH1 ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRV 113 G4CH1 ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRV 114

Table 24
Amino acid sequences of CH2 Fc Combination code Amino acid sequences of CH2 SEQ
ID NO. G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK 115 G2CH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTK 116 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 G3CH2 CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKTK 118 G4CH2 APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAK 119

Table 25
Amino acid sequences of CH3 Fc forming a heterodimer Combination code Amino acid sequences of CH3 SEQ
ID NO. G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 120 G2CH3 GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 121 G3CH3 GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGK 122 G4CH3 GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 123 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 124 G2KCH3 GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 125 G3KCH3 GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPG 126 G4KCH3 GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG 127 G1ACH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGA 128

Specific antigen sequences

Table 26
Amino acid sequences of tumor antigen Name of tumor antigens (source) Amino acid sequences SEQ
ID NO. human CD38
(source: UniProtKB - P28907) VPRWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEAACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEK VQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIYRPDKFLQCVKNPEDSSCTSEI 129 Human BCMA
(source: UniProtKB - Q02223) MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNA 130 CTLA-4 people
(source: UniProtKB - P16410) MHVAQPAVVLASSRGIASFVCEYASPGKATEVRVTVLRQADSQVTEVCAATYMMGNELTFLDSICTGTSSGNQVNLTIQGLRAMDTGLYICKVELMYPPPYYLGIGNGTQIYVIDPEPCPDSD 131 LAG-3 people
(source: UniProtKB - P18627) VPVVWAQEGAPAQLPCSPTIPLQDLSLLRRAGVTWQHQPDSGPPAAAPGHPLAPGPHPAAPSSWGPRPRRYTVLSVGPGGLRSGRLPLQPRVQLDERGRQRGDFSLWLRPARRADAGEYRAAVHLRDRALSCRLRRLGQASMTASPPGSLRASDWVILNCSFSRPDRPASVHWFRNRGQGRVPVRESPHHHLAESFLFLPQVSPMDS GPWGCILTYRDGFNVSIMYNLTVLGLEPPTPLTVYAGAGSRVGLPCRLPAGVGTRSFLTAKWTPPGGGPDLLVTGDNGDFTLRLEDVSQAQAGTYTCHIHLQEQQLNATVTLAIITVTPKSFGSPGSLGKLLCEVTPVSGQERFVWSSLDTPSQRSFSGPWLEAQEAQLLSQPWQCQLYQGERLLGAAVYFTELSSPGAQRSGR APGALPAGHL 132 Human TIGIT
(source: UniProtKB - Q495A1) MMTGTIETTGNISAEKGGSIILQCHLSSTTAQVTQVNWEQQDQLLAICNADLGWHISPSFKDRVAPGPGLGLTLQSLTVNDTGEYFCIYHTYPDGTYTGRIFLEVLESSVAEHGARFQIP 133 human PD-1
(source: UniProtKB - Q15116) PGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSNTSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRNDSGTYLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLV 134 Human PD-L1
(source: UniProtKB - Q9NZQ7) FTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIF YCTFRRLDPEENHTAELVIPELPLAHPPNER 135 Human SLAMF7
(source: UniProtKB - Q9NQ25) SGPVKELVGSVGGAVTFPLKSKVKQVDSIVWTFNTTPLVTIQPEGGTIIVTQNRNRERVDFPDGGYSLKLSKLKKNDSGIYYVGIYSSSLQQPSTQEYVLHVYEHLSKPKVTMGLQSNKNGTCVTNLTCCMEHGEEDVIYTWKALGQAANESHNGSILPISWRWGESDMTFICVARNPVSRNFSSPILARK LCEGAADDPDSSM 136 Human CEA
(source: UniProtKB - P06731) KLTIESTPFNVAEGKEVLLLVHNLPQHLFGYSWYKGERVDGNRQIIGYVIGTQQATPGPAYSGREIIYPNASLLIQNIIQNDTGFYTLHVIKSDLVNEEATGQFRVYPELPKPSISSNNSKPVEDKDAVAFTCEPETQDATYLWWVNNQSLPVSPRLQLSNGNRTLTLFNVTRNDTASYKCETQNPVSARRSD SVILNVLYGPDAPTISPLNTSYRSGENLNLSCHAASNPPAQYSWFVNGTFQQSTQELFIPNITVNNSGSYTCQAHNSDTGLNRTTVTTITVYAEPPKPFITSNNSNPVEDEDAVALTCEPEIQNTTYLWWVNNQSLPVSPRLQLSNDNRTLTLLSVTRNDVGPYECGIQNKLSVDHSDPVILNVLYGPDDPTISPSYTYYRPGVNLSLSCHAASNPPA QYSWLIDGNIQQHTQELFISNITEKNSGLYTCQANNSASGHSRTTVKTITVSAELPKPSISSNNSKPVEDKDAVAFTCEPEAQNTTYLWWVNGQSLPVSPRLQLSNGNRTLTLFNVTRNDARAYVCGIQNSVSANRSDPVTLDVLYGPDTPIISPPDSSYLSGANLNLSCHSASNPSPQYSWRINGIPQQHTQVLFIAKITPNNNGTYACF VSNLATGRNNSIVKSITVSASGTSPGLSA 137 Human CD3ε
(source: UniProtKB - P07766) DGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILWQHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARVCENCMEMD 138 Human CD16A
(source: UniProtKB - P08637) GMRTEDLPKAVVFLEPQWYRVLEKDSVTLKCQGAYSPEDNSTQWFHNESLISSQASSYFIDAATVDDSGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRKYFHHNSDFYIPKATLKDSGSYFCRGLFGSKNVSSETVNITITQGLAVSTISSFFPPGY Q 139 Human TGF-β1
(source: UniProtKB - P01137) ALDTNYCFSSTEKNCCVRQLYIDFRKDLGWKWIHEPKGYHANFCLGPCPYIWSLDTQYSKVLALYNQHNPGASAAPCCVPQALEPLPIVYYVGRKPKVEQLSNMIVRSCKCS 140 Human TGF-β2
(source: UniProtKB - P61812) ALDAAYCFRNVQDNCCLRPLYIDFKRDLGWKWIHEPKGYNANFCAGACPYLWSSDTQHSRVLSLYNTINPEASASPCCVSQDLEPLTILYYIGKTPKIEQLSNMIVKSCKCS 141 Human TGF-β3
(source: UniProtKB - P10600) ALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTTHSTVLGLYNTLNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLSNMVVKSCKCS 142 Human VEGFA
(source: UniProtKB - P15692) APMAEGGGQNHHEVVKFMDVYQRSYCHPIETLVDIFQEYPDEIEYIFKPSCVPLMRCGGCCNDEGLECVPTEESNITMQIMRIKPHQGQHIGEMSFLQHNKCECRPKKDRARQEKKSVRGKGKGQKRKRKKSRYKSWSVYVGARCCLMPWSLPGPHPCGPCSERRKHLFVQDPQTCKCSCKNTDSRCKARQLEL NERTCRCDKPRR 143 Human IL-10
(source: UniProtKB - P22301) PGQGTQSENSCTHFPGNLPNMLRDLRDAFSRVKTFFQMKDQLDNLLLKESLLEDFKGYLGCQALSEMIQFYLEEVMPQAENQDPDIKAHVNSLGENLKTLRLRLRRCHRFLPCENKSKAVEQVKNAFNKLQEKGIYKAMSEFDIFINYIEAYMTMKIRN 144 Human IL-10R
(source: UniProtKB - Q13651) HGTELPSPPSVWFEAEFFHHILHWTPIPNQSESTCYEVALLRYGIESWNSISNCSQTLSYDLTAVTLDLYHSNGYRARVRAVDGSRHSNWTVTNTRFSVDEVTLTVGSVNLEIHNGFILGKIQLPRPKMAPANDTYESIFSHFREYEIAIRKVPGNFTFTHKKVKHENFSLLTSGEVGEFCVQVKPSVASRSNKGMWSKEE CISLTRQYFTVTN 145

Preparation of Antibody and Detection of Antibody Activity

Example 1: Preparation of a bispecific antibody

1. Method for constructing a plasmid

pcDNA3.1 (purchased from invitrogen) or pCHO1.0 (purchased from Gibco) was used as the vector.

1.1 Amplification of a DNA fragment of interest using polymerase chain reaction (PCR)

PCR system (µl) 50µl DNA polymerase (2× PrimeSTAR Max Premix TaKaRa Item No. R405A) 25µl Forward primer (10 µM) 1µl Reverse primer (10µM) 1µl Template DNA* 1µl _H2O 22µl *Template DNA synthesized by Wuhan Genecreate. The DNA sequence was obtained by reverse translation based on the amino acid sequences SEQ ID NO 1-145

PCR program Temperature Time The number of cycles 95°С 5 minutes 1 98°С 10sec 25-30 55°C 5 sec 72°C 12kb/min 4°C ∞ 1

The resulting PCR product represents the DNA fragment of interest.

1.2 Restriction enzyme digestion of vector plasmid

Vector plasmid DNA* 10mcg buffer 10µl Restriction enzyme (such as NotI NruI or BamHI-HF, etc.) 5µl _H2O Add up to 100µl Overall volume 100µl

Enzymatic digestion is carried out at the optimal temperature for 4 hours.

The enzyme-cleaved product is an enzyme-cleaved vector DNA.

*Vectors are divided into two types: heavy chain expression vector and light chain expression vector. The heavy chain expression vector contains a signal peptide and a DNA sequence of the human IgG1 heavy chain constant region (GenBank accession number MG920253.1), which includes CH1, hinge, CH2 and CH3; and the restriction site is located between the 3' signal peptide and the 5' end of CH1; wherein the light chain expression vector contains a signal peptide and a human kappa or lambda light chain constant region DNA sequence, and a restriction site is located between the 3' signal peptide and the 5' end of the light chain constant region.

1.3 Purification of PCR products or enzyme-digested products

Tiangen reagents were used for DNA purification. For specific steps, see the instructions included with the Tiangen kit. The resulting purified product was a purified DNA fragment of interest and a purified enzyme-digested vector DNA.

(1) Recombination of fragments of interest

Buffer 5XCE II 4 µl Exnase II Recombinase, Vazyme, Item No. C112-01 2 µl Enzyme-split vector 50-200 ng Fragment 1 20-200 ng Fragment 2 20-200 ng _H2O up to 20µl

Recombination conditions: 37°C for 30 minutes. The resulting recombinant product was placed on ice and was ready for transformation.

The heavy chain fragment was recombined into an enzyme-digested vector DNA for heavy chain expression, and the light chain fragment was recombined into an enzyme-digested vector DNA for light chain expression.

1.4 Transformation by heat shock method

10 μl of the recombinant product was transformed into 100 μl of Trans10-competent cells by heat shock according to the conventional method. A single colony was selected and sequenced by Wuhan Genecreate. The plasmid produced by recombination of the heavy chain fragment into the enzyme-digested DNA of the expression vector is called the heavy chain expression plasmid, and the plasmid obtained by recombination of the light chain fragment into the enzyme-digested DNA of the expression vector is called the light chain expression plasmid, and the plasmid obtained by recombination of a hybrid peptide fragment into enzyme-digested DNA of an expression vector is called a hybrid peptide expression plasmid, a plasmid obtained by recombination of a fragment of a hybrid heavy chain into enzyme-digested DNA of an expression vector is called a hybrid heavy chain expression plasmid, and a plasmid obtained by recombination of a fragment of a hybrid chain into an enzyme-digested expression vector, called a hybrid chain expression plasmid.

The design of the specific plasmid is as follows:

1) Structure 1 of bispecific antibody in FIG. 1 includes a two-plasmid construct. The two plasmids are: a hybrid light chain expression plasmid (pFL) and a hybrid heavy chain expression plasmid (pFH).

2) Structure 2 of bispecific antibody in FIG. 2 includes the construction of their two plasmids. The two plasmids are: light chain expression plasmid (pL) and hybrid heavy chain 2 expression plasmid (pFH2).

3) Structure 3 of bispecific antibody in FIG. 3 includes a single plasmid construct. This plasmid is the following: fusion peptide expression plasmid (pFP).

4) Structure 4 of the bispecific antibody in FIG. 4 includes a two-plasmid construct. The two plasmids are: hybrid light chain expression plasmid 2 (pFL2) and hybrid heavy chain expression plasmid 3 (pFH3).

1.5 Method of expression of bispecific antibody

Two transient transfection expression systems are involved, CHAOS and 293E, and the steps are detailed below:

(1) CHO-S transient transfection procedure (100ml total transfection volume as an example)

a) Cell passage was carried out one day before transfection, for example, CD-CHO medium (purchased from Thermo Fisher) can be used for cell passage; the cell density in the suspension was adjusted to 1×10 ⁶ cells/ml, and the volume was 90 ml to ensure that the cells were in the logarithmic growth phase, and the cell density could reach 2×10 ⁶ cells/ml on the next day of transfection;

b) cells were cultured at 37°C, 125 rpm, 5% CO ₂ overnight on a shaker;

c) on the day determined for transfection, FectoPRO transfection reagent (purchased from Polyplus transfection company) was prewarmed to room temperature and mixed gently; 50 μg of plasmid DNA was diluted in 10 ml of serum-free medium such as opti PRO-SFM (purchased from Thermo Fisher), mixed gently and added to 100 μl of transfection reagent, mixed thoroughly and incubated at room temperature for 10 minutes to form a transfection complex; two or three plasmid DNAs were cotransfected during the transfection process.

d) the transfection complex was uniformly added to the resulting 90 ml of logarithmic growth phase cells, vortexed immediately after mixing and placed on a culture shaker at 37°C, 125 rpm, 5% CO ₂ ;

e) 2-4 hours after transfection, 75 μl of Fecto PRO® Booster transfection enhancer (purchased from Polyplus transfection company) was added;

f) 18-24 hours after transfection, the cells were cooled to 32°C and incubated;

g) feeding was carried out on days 3, 5 and 7 after transfection, and the feeding volume was 3.5% of the total cell volume;

h) cells were harvested when cell viability was less than 70% and expression time was 9-13 days.

(2) 293E transient transfection procedure (20 ml total transfection volume as an example)

a) Cell passage was carried out one day before transfection, for example, FreeStyle™ 293 (purchased from Thermo Fisher) can be used for cell passage; The cell density in the suspension was adjusted to 0.6×10 ⁶ -0.8×10 ⁶ cells/ml, and the volume was 20 ml to ensure that the cells were in the logarithmic growth phase, and the cell density could reach 1.2×10 ⁶ -1.6×10 ⁶ cells/ml on the next day of transfection;

b) cells were cultured at 37°C, 125 rpm, 5% CO ₂ overnight on a shaker;

c) on the day determined for transfection, LPEI was prewarmed to room temperature and mixed gently before use;

d) 20 µg DNA was diluted in 0.67 ml serum-free medium such as FreeStyle™ 293 and mixed thoroughly;

e) 40 µg LPEI was diluted with 0.67 ml of serum-free medium such as FreeStyle™ 293 and mixed thoroughly;

f) the diluted LPEI obtained in step e) was added to the diluted DNA obtained in step d), quickly mixed thoroughly and incubated at room temperature for 15 minutes to form a complex;

g) the transfection complex obtained in step f) was uniformly added to the resulting 20 ml of logarithmic growth phase cells, vortexed immediately after mixing and placed on a culture shaker at 37°C, 125 rpm, 5% CO ₂ ;

h) feeding was carried out on the first and third days after transfection, and the feeding volume was 5% of the total cell volume;

i) cells were expressed until day six and then harvested.

(3) The specific cotransfected plasmid DNA was as follows:

1) To express bispecific antibody 1 shown in FIG. 1, plasmids pFL and pFH were required for cotransfection into CHO-S or 293E cells for expression;

2) To express bispecific antibody 2 shown in FIG. 2, plasmids pL and pFH2 were required for cotransfection into CHO-S or 293E cells for expression;

3) To express bispecific antibody 3 shown in FIG. 3, pFP plasmid was required for cotransfection into CHO-S or 293E cells for expression;

4) To express bispecific antibody 4 shown in FIG. 4, plasmids pFL2 and pFH3 were required for cotransfection into CHO-S or 293E cells for expression.

Typically, if two plasmids are cotransfected for expression, the molar ratio of the two plasmids can be 1:1 or any other ratio.

2. Method for purifying a bispecific antibody:

The antibody purification method preferably includes affinity chromatography, ion exchange chromatography, hydrophobic interaction chromatography and molecular sieves, and the steps are as follows:

(1) the culture solution of antibody-expressing cells was collected and centrifuged at 3000×g for 10 minutes, then the supernatant was collected, filtered through a 0.22 μm filter and stored at 4°C for later use;

(2) affinity chromatography (MabSelect SuRe GE 17-5438-01, 18ml column volume as an example)

a) equilibration: the column was equilibrated with binding buffer (25 mM Tris, pH 7.0-7.4) until UV detection and conductivity were stable or at baseline, using at least five column volumes to balancing;

b) sample loading: the filtered supernatant obtained in step (1) was loaded at a flow rate of 5 ml/min;

c) washing and equilibration: five column volumes of binding buffer were used for washing;

d) elution: the sample was eluted with elution buffer (50 mM citric acid, pH 3.4 ± 0.1) at a flow rate of 5 ml/min, 5 column volumes were used for elution and the eluted peak was collected;

e) Neutralization: The eluent was neutralized with 1M Tris pH8.0 and the sample pH was adjusted to 6.0±0.1.

(3) ion exchange chromatography (cation exchange chromatography as an example, HiTrap SP-HP GE 17-1151-01 5ml column volume)

a) sample preparation: the affinity chromatography sample was subjected to microfiltration and then diluted with ultrapure water to make the conductivity less than 5 mS/cm, and then the pH was adjusted to 6.0 ± 0.1;

b) Equilibration and sample loading: The column was first equilibrated with 5 column volumes of buffer B (25mM citric acid + 1M sodium chloride, whose conductivity should be 80 ~ 90 mS/cm, pH 6.0 ± 0.1), and then equilibrated at less at least five column volumes of buffer A (25 mM citric acid, conductivity less than 5 mS/cm, pH 6.0 ± 0.1) until conductivity, pH, and baseline UV level were stable, and then loaded sample at a flow rate of 3 ml/min;

c) washing and equilibration: the column was washed with five column volumes of buffer A at a flow rate of 5 ml/min;

d) elution: the sample was eluted with 20 column volumes of 0-30% buffer B, followed by 10 column volumes of 100% buffer B; The flow rate throughout the process was 3 ml/min, and the eluent was collected in separate tubes and tested.

(4) hydrophobic chromatography (Capto phenyl ImpRes filler GE XK16/20 11.5cm/23ml)

a) sample treatment: 5M sodium chloride was added to the sample with 1M sodium chloride and the pH was adjusted to 6.0;

b) equilibration and sample loading: first, the column was equilibrated with five column volumes of buffer A (25 mM citrate + 1 M sodium chloride, pH 6.0 ± 0.1) at a flow rate of 5 ml/min; and the sample was loaded at a flow rate of 3.3 ml/min;

c) washing and equilibration: five column volumes of buffer A were used for washing at a flow rate of 5 ml/min; For washing, five column volumes of 10% buffer B (25 mM citrate, pH 6.0 ± 0.1) were used at a flow rate of 5 ml/min;

d) elution: 90% buffer B was used for elution at a flow rate of 5 ml/min, the eluted peaks were collected in separate tubes and then detected;

(5) molecular sieve (HiLoad Superdex 200pg GE 28989336 26/600)

a) equilibration and sample loading: the column was equilibrated with two column volumes of buffer (20 mM histidine + 0.15 M sodium chloride, pH 6.0 ± 0.1), and then loaded the sample at a flow rate of 3 ml/min;

b) Elution: Two column volumes of buffer were used for elution, and the eluted peaks were collected in separate tubes and then detected.

The antibody codes of some specifically expressed antibodies and the corresponding amino acid sequences of the antibodies are shown in the following tables :

Table 27
Antibody codes and amino acid sequences of some antibodies with the structure of 1 bispecific antibody Antibody code Peptides Domain Code Amino acid sequences (bold and underlined amino acids represent CDRs) SEQ ID NO. Y100-A1 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin14 GGGGSGGGGSGGGGS 82 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin14 GGGGSGGGGSGGGGS 82 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A2 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A3 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A4 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 12A4 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
SNWPTFGQGTKVEIK 46 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQ
GLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRS
EDTAVYFCARKFHFVSSGSPFGMDVWGQGTTVTVSS 45 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A5 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 12A4 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
SNWPTFGQGTKVEIK 46 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQ
GLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRS
EDTAVYFCARKFHFVSSGSPFGMDVWGQGTTVTVSS 45 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A6 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A7 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A8 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A9 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A10 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb Avastin DIQMTQSPSSLSSASVGDRVTITCSASQDISNYLNWYQQKPGKAP
KVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ
YSTVPWTFGQGTKVEIK 54 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb Avastin EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPG
KGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNS
LRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSS 53 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-A11 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-AC1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-AC2 Hybrid light chain VLa 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-AC3 Hybrid light chain VLa 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-A1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-A2 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-A3 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-A4 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-A5 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-A1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-A2 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-A3 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-A1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-A2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-A3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-A1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-A2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-A3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-A1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-A2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-A3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-A4 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-A5 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-A1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKIRRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-A2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-A3 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-A4 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-A1 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-A2 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-A3 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B1 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin14 GGGGSGGGGSGGGGS 82 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin14 GGGGSGGGGSGGGGS 82 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Linker 3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B2 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B3 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B4 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQ
GLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRS
EDTAVYFCARKFHFVSSGSPFGMDVWGQGTTVTVSS 45 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb 12A4 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
SNWPTFGQGTKVEIK 46 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B5 Hybrid light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTYAISWVRQAPGQ
GLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRS
EDTAVYFCARKFHFVSSGSPFGMDVWGQGTTVTVSS 45 Hybrid heavy chain VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 12A4 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPR
LLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQR
SNWPTFGQGTKVEIK 46 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B6 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B7 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B8 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B9 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B10 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb Avastin EVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPG
KGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNS
LRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSS 53 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb Avastin DIQMTQSPSSLSSASVGDRVTITCSASQDISNYLNWYQQKPGKAP
KVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQ
YSTVPWTFGQGTKVEIK 54 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B11 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin16 EAAAKGGGGSGGGGS 84 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin16 EAAAKGGGGSGGGGS 84 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B12 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin18 EAAAAKEAAAKGGGGS 86 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin18 EAAAAKEAAAKGGGGS 86 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-B13 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-BC1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-BC2 Hybrid light chain VLa 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y100-BC3 Hybrid light chain VLa 4420 dvvmtqtplslpvslgdqasisc rssqslvhsngntylr wylq
kpgqspkvliy kvsnrfs gvpdrfsgsgsgtdftlkisrveaedl
gvyfc sqsthvpwt fgggtkleik 68 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 4420 evkldetggglvqpgRPMklscvasgftfs dywmn wvrqspe
kglewva qirnkpynyetyysdsvkg rftisrddskssvylqm
nnlrvedmgiyyctg syygmdy wgqgtsvtvss 67 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-B1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-B2 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-B3 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-B4 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y101-B5 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin16 EAAAKGGGGSGGGGS 84 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin16 EAAAKGGGGSGGGGS 84 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 120 Y101-B6 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin18 EAAAAKEAAAKGGGGS 86 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin18 EAAAAKEAAAKGGGGS 86 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-B1 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-B2 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin18 EAAAAKEAAAKGGGGS 86 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin18 EAAAAKEAAAKGGGGS 86 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y103-B3 Hybrid light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin18 EAAAAKEAAAKGGGGS 86 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTA
ADTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 Hybrid heavy chain VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGK
GLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin18 EAAAAKEAAAKGGGGS 86 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPLT FGQGTNLEIK 34 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-B1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-B2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y104-B3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-B1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-B2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y105-B3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSPGK
GLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQMNDL
KSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-B1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-B2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKA
PKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC Q
QGYGNPFT FGQGTKVEIK 58 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-B3 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-B4 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y106-B5 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKASGYTFSSNVISWVRQAPGQ
GLEWMGGVIPIVDIANYAQRFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCASTLGLVLDAMDYWGQGTLVTVSS 59 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb 3G12 ETVLTQSPGTLLSLSPGERATLSCRASQSLGSSYLAWYQQKPGQA
PRLLIYGASSRAPGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQ
QYADSPITFGQGTRLEIK 60 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-B1 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-B2 Hybrid light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
SSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Hybrid heavy chain VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-B3 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-B4 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y110-B5 Hybrid light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAP
RLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQ
RSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQA
DDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Hybrid heavy chain VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQ
GLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSLR
SEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLG
VYYC FQGSHVPWT FGGGTKLEIK 64 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-B1 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin21 GAAAGGGGSGGGGSGGGGS 89 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin15 GAAAGGGGSGGGGS 83 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-B2 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y116-B3 Hybrid light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Linker 1 Lin17 GGGGSEAAAKGGGGS 85 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Hybrid heavy chain VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Linker 2 Lin17 GGGGSEAAAKGGGGS 85 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 Linker 3 Lin6 GYPGGGGS 74 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE
VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWL
NGKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120

Table 28
Antibody codes and amino acid sequences of some antibodies with bispecific antibody structure 2 for comparison Antibody code Polypeptides Domain Code Amino acid sequences (bold and underlined amino acids represent CDRs) SEQ ID NO. Y200-A1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 Y200-A2 Light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQRSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQA
PGQGLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAY
MELSSLRSEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTV
TVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb Avastin EVQLVESGGGLVQPGGSLRLSCAAS GYTFTNYGMN WVRQ
APGKGLEWVG WINTYTGEPTYAADFKR RFTFSLDTSKSTA
YLQMNSLRAEDTAVYYCAK YPHYYGSSHWYFDV WGQGT
LVTVSS 53 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb Avastin DIQMTQSPSSLSSASVGDRVTITC SASQDISNYLN WYQQKPG
KAPKVLIY FTSSLHS GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYC QQYSTVPWT FGQGTKVEIK 54 Y200-A3 Light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 Y201-A1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS WVRQA
PGQGLEWMG GVIPIVDIANYAQ RFKGRVTITADESTSTTY
MELSSLRSEDTAVYYCA STLGLVLDAMDY WGQGTLVTVS
S 59 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb 3G12 ETVLTQSPGTLSLSPGERATLSC RASQSLGSSYLA WYQQKP
GQAPRLLIY GASSRAP GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYADSPIT FGQGTRLEIK 60 Y201-A2 Light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQRSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQA
PGQGLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAY
MELSSLRSEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTV
TVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS WVRQA
PGQGLEWMG GVIPIVDIANYAQ RFKGRVTITADESTSTTY
MELSSLRSEDTAVYYCA STLGLVLDAMDY WGQGTLVTVS
S 59 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb 3G12 ETVLTQSPGTLSLSPGERATLSC RASQSLGSSYLA WYQQKP
GQAPRLLIY GASSRAP GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYADSPIT FGQGTRLEIK 60 Y203-A1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQ
PPGKGLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSL
KLRSVTAADTAVYYCA FGYSDYEYNWFDP WGQGTLVTV
SS 33 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQ
APRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAV
YYC QQRSNWPLT FGQGTNLEIK 34 Y204-A1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQ
APGKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTL
YLQMNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKP
GQAPRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYGSSPWT FGQGTKVEIK 28 Y205-A1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSP
GKGLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQ
MNDLKSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA
WYQQKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTIT
SVQAEDMGQYFC QQGINNPLT FGDGTKLEIK thirty Y205-A2 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSP
GKGLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQ
MNDLKSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA
WYQQKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTIT
SVQAEDMGQYFC QQGINNPLT FGDGTKLEIK thirty Y206-A1 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 Y206-A2 Light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 Y210-A1 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSP
GKGLEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFK
MNSLQADDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE W
YLQKPGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITR
LEAEDLGVYYC FQGSHVPWT FGGGTKLEIK 64 Y216-A1 Light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPG
KVPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDV
ATYYC QQYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQ
APGKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQ
MNSLRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQ
APGKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQ
APVLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEA
DYYC NSRDSSGNHVV FGGGTKLTVL 48 Y200-B1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 Y200-B2 Light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQRSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQA
PGQGLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAY
MELSSLRSEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTV
TVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb Avastin DIQMTQSPSSLSSASVGDRVTITC SASQDISNYLN WYQQKPG
KAPKVLIY FTSSLHS GVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYC QQYSTVPWT FGQGTKVEIK 54 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb Avastin EVQLVESGGGLVQPGGSLRLSCAAS GYTFTNYGMN WVRQ
APGKGLEWVG WINTYTGEPTYAADFKR RFTFSLDTSKSTA
YLQMNSLRAEDTAVYYCAK YPHYYGSSHWYFDV WGQGT
LVTVSS 53 Y200-B3 Light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Y201-B1 Light chain VLa 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQRSNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQA
PGQGLEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAY
MELSSLRSEDTAVYFCAR KFHFVSGSPFGMDV WGQGTTV
TVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb 3G12 ETVLTQSPGTLSLSPGERATLSC RASQSLGSSYLA WYQQKP
GQAPRLLIY GASSRAP GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYADSPIT FGQGTRLEIK 60 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS WVRQA
PGQGLEWMG GVIPIVDIANYAQ RFKGRVTITADESTSTTY
MELSSLRSEDTAVYYCA STLGLVLDAMDY WGQGTLVTVS
S 59 Y201-B2 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb 3G12 ETVLTQSPGTLSLSPGERATLSC RASQSLGSSYLA WYQQKP
GQAPRLLIY GASSRAP GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYADSPIT FGQGTRLEIK 60 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS WVRQA
PGQGLEWMG GVIPIVDIANYAQ RFKGRVTITADESTSTTY
MELSSLRSEDTAVYYCA STLGLVLDAMDY WGQGTLVTVS
S 59 Y203-B1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQ
APRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAV
YYC QQRSNWPLT FGQGTNLEIK 34 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQ
PPGKGLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSL
KLRSVTAADTAVYYCA FGYSDYEYNWFDP WGQGTLVTV
SS 33 Y204-B1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKP
GQAPRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDF
AVYYC QQYGSSPWT FGQGTKVEIK 28 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQ
APGKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTL
YLQMNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Y205-B1 Light chain VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQYLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQA
PGKGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYL
QMNSLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA
WYQQKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTIT
SVQAEDMGQYFC QQGINNPLT FGDGTKLEIK thirty Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSP
GKGLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQ
MNDLKSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Y205-B2 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA
WYQQKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTIT
SVQAEDMGQYFC QQGINNPLT FGDGTKLEIK thirty Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb 10A7 EVQLVESGGGLTQPGKSLKLSCEASGFTF SSFTMH WVRQSP
GKGLEWVAFI RSGSGIVFYADAVRG RFTISRDNAKNLLFLQ
MNDLKSEDTAMYYCAR RPLGHNTFDS WGQGTLVTVSS 29 Y206-B1 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 Y206-B2 Light chain VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKP
GKAPKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDF
ATYYC QQGYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQA
PGKGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAY
LQMNSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVS
S 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Y210-B1 Light chain VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPG
QAPRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFA
VYYC QQSSNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQ
APGKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNT
LFLQMNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE W
YLQKPGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITR
LEAEDLGVYYC FQGSHVPWT FGGGTKLEIK 64 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSP
GKGLEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFK
MNSLQADDTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 Y216-B1 Light chain VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPG
KVPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDV
ATYYC QQYSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW
KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain2 VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQ
APGKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQ
MNSLRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW
NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEA
PEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVH
QDWLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1KCH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEW
ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 124 Linker 4 Lin19 AGGGGSGGGGSGGGGSG 87 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQ
APVLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEA
DYYC NSRDSSGNHVV FGGGTKLTVL 48 Linker 5 Lin14 GGGGSGGGGSGGGGS 82 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQ
APGKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNS
LYLQMNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47

Table 29
Antibody codes and amino acid sequences of some antibodies with bispecific antibody structure 3 for comparison Antibody code Polypeptides Domain Code Amino acid sequences (bold and underlined amino acids represent CDRs) SEQ ID NO. Y300-A1 Hybrid peptide VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPG
KGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMN
SLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQYLYHPAT FGQGTKVEIK 42 Linker 7 Lin5 GGGGSAS 73 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPG
KGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQM
NSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQGYGNPFT FGQGTKVEIK 58 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y304-A1 Hybrid peptide VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAP
GKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQ
MNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQA
PRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYY
C QQSSNWPRT FGQGTKVEIK 38 Linker 7 Lin5 GGGGSAS 73 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAP
GKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQ
MNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQ
APRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVY
YC QQYGSSPWT FGQGTKVEIK 28 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y316-A1 Hybrid peptide VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAP
GKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNS
LRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGK
VPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATY
YC QQYSSYPYT FGQGTKVEIK 50 Linker 7 Lin5 GGGGSAS 73 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAP
GKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQ
MNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAP
VLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYY
C NSRDSSGNHVV FGGGTKLTVL 48 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y300-B1 Hybrid peptide VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPG
KGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMN
SLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQYLYHPAT FGQGTKVEIK 42 Linker 7 Lin5 GGGGSAS 73 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQGYGNPFT FGQGTKVEIK 58 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPG
KGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQM
NSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y304-B1 Hybrid peptide VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAP
GKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQ
MNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQA
PRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYY
C QQSSNWPRT FGQGTKVEIK 38 Linker 7 Lin5 GGGGSAS 73 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQ
APRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVY
YC QQYGSSPWT FGQGTKVEIK 28 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAP
GKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQ
MNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y316-B1 Hybrid peptide VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAP
GKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNS
LRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGK
VPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATY
YC QQYSSYPYT FGQGTKVEIK 50 Linker 7 Lin5 GGGGSAS 73 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAP
VLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYY
C NSRDSSGNHVV FGGGTKLTVL 48 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAP
GKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQ
MNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y300-C1 Hybrid peptide VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQYLYHPAT FGQGTKVEIK 42 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPG
KGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMN
SLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 7 Lin5 GGGGSAS 73 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPG
KGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQM
NSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQGYGNPFT FGQGTKVEIK 58 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y304-C1 Hybrid peptide VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQA
PRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYY
C QQSSNWPRT FGQGTKVEIK 38 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAP
GKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQ
MNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 7 Lin5 GGGGSAS 73 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAP
GKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQ
MNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQ
APRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVY
YC QQYGSSPWT FGQGTKVEIK 28 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y316-C1 Hybrid peptide VLa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAP
GKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNS
LRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGK
VPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATY
YC QQYSSYPYT FGQGTKVEIK 50 Linker 7 Lin5 GGGGSAS 73 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAP
GKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQ
MNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAP
VLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYY
C NSRDSSGNHVV FGGGTKLTVL 48 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y300-D1 Hybrid peptide VLa S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQYLYHPAT FGQGTKVEIK 42 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPG
KGLEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMN
SLRAEDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 7 Lin5 GGGGSAS 73 VLb G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGK
APKLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYY
C QQGYGNPFT FGQGTKVEIK 58 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPG
KGLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQM
NSLRAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y304-D1 Hybrid peptide VLa 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQA
PRLLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYY
C QQSSNWPRT FGQGTKVEIK 38 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAP
GKGLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQ
MNSLRAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 7 Lin5 GGGGSAS 73 VLb Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQ
APRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVY
YC QQYGSSPWT FGQGTKVEIK 28 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAP
GKGLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQ
MNSLRAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120 Y316-D2 Hybrid peptide VLa Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGK
VPKLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATY
YC QQYSSYPYT FGQGTKVEIK 50 Linker 6 Lin14 GGGGSGGGGSGGGGS 82 VHa Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAP
GKGLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNS
LRAEDTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 7 Lin5 GGGGSAS 73 VLb NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAP
VLVIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYY
C NSRDSSGNHVV FGGGTKLTVL 48 Linker 8 Lin14 GGGGSGGGGSGGGGS 82 VHb NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAP
GKGLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQ
MNSLRAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPE
VQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQD
WLNGKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES
NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSC
SVMHEALHNHYTQKSLSLSPGK 120

Table 30
Antibody codes and amino acid sequences of some antibodies with bispecific antibody structure 4 for comparison Antibody code Polypeptide Domain Code Amino acid sequences (bold and underlined amino acids represent CDRs) SEQ ID NO. Y400-A1 Hybrid light chain VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
YLYHPAT FGQGTKVEIK 42 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
GYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGKG
LEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSLRA
EDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y404-A1 Hybrid light chain VLb 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQS
SNWPRT FGQGTKVEIK 38 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VHb 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y416-A1 Hybrid light chain VLb Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VHb Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y400-B1 Hybrid light chain VHb S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGKG
LEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSLRA
EDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
GYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VLb S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
YLYHPAT FGQGTKVEIK 42 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGK
GLEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSL
RAEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y404-B1 Hybrid light chain VHb 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQA
PRLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Q
QYGSSPWT FGQGTKVEIK 28 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VLb 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQS
SNWPRT FGQGTKVEIK 38 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSL
RAEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120 Y416-B1 Hybrid light chain VHb Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 Linker 10 Lin14 GGGGSGGGGSGGGGS 82 VLa NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Hybrid heavy chain VLb Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC Q
QYSSYPYT FGQGTKVEIK 50 Linker 9 Lin14 GGGGSGGGGSGGGGS 82 VHa NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKP
SNTKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEV
QFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLN
GKEYKCKVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESN
GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV
MHEALHNHYTQKSLSLSPGK 120

Table 31
Antibody codes and amino acid sequences of monoclonal antibodies Antibody code Polypeptides Domain Code Amino acid sequences (bold and underlined amino acids represent CDRs) SEQ ID NO. S70mAb Light chain VL S70 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
YLYHPAT FGQGTKVEIK 42 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH S70 EVQLVESGGGLVQPGGSLRLSCAASGFTF SDSWIH WVRQAPGKG
LEWVAWI SPYGGSTYYADSVKG RFTISADTSKNTAYLQMNSLRA
EDTAVYYCAR RHWPGGFDY WGQGTLVTVSS 41 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 12A4mAb Light chain VL 12A4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQR
SNWPT FGQGTKVEIK 46 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH 12A4 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFS TYAIS WVRQAPGQG
LEWMGGI IPIFGKAHYAQKFQG RVTITADESTSTAYMELSSSLRSE
DTAVYFCAR KFHFVSGSPFGMDV WGQGTTVTVSS 45 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 3G12mAb Light chain VL 3G12 ETVLTQSPGTLSLSPGERATLSC RASQSLGSSYLA WYQQKPGQAP
RLLIY GASSRAP GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQ
YADSPIT FGQGTRLEIK 60 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH 3G12 QVQLVQSGAEVKKPGSSVKVSCKAS GYTFSSNVIS WVRQAPGQ
GLEWMG GVIPIVDIANYAQ RFKGRVTITADESTSTTYMELSSLRS
EDTAVYYCA STLGLVLDAMDY WGQGTLVTVSS 59 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 LAG35mAb Light chain VL LAG35 EIVLTQSPATLSLSPGERATLSC RASQSISSYLA WYQQKPGQAPRL
LIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQRS
NWPLT FGQGTNLEIK 34 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH LAG35 QVQLQQWGAGLLKPSETLSLTCAVYGG SFSDYYWN WIRQPPGK
GLEWIGEI NHRGSTNSNPSLKS RVTLSLDTSKNQFSLKLRSVTAA
DTAVYYCA FGYSDYEYNWFDP WGQGTLVTVSS 33 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 5C4mAb Light chain VL 5C4 EIVLTQSPATLSLSPGERATLSC RASQSVSSYLA WYQQKPGQAPR
LLIY DASNRAT GIPARFSGSGSGTDFTLTISSLEPEDFAVYYC QQS
SNWPRT FGQGTKVEIK 38 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH 5C4 QVQLVESGGGVVQPGRSLRLDCKASGITFS NSGMH WVRQAPGK
GLEWVAVI WYDGSKRYYADSVKG RFTISRDNSKNTLFLQMNSL
RAEDTAVYYCA TNDDY WGQGTLVTVSS 37 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 10A7mAb Light chain VL 10A7 DIVMTQSPSSLAVSPGEKVTMTC KSSQSLYYSGVKENLLA WYQ
QKPGQSPKLLIY YASIRFT GVPDRFTGSGSGTDYTLTITSVQAED
MGQYFC QQGINNPLT FGDGTKLEIK thirty C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 BN10mAb Light chain VL B-N10 DVLMTQTPLSLPVSLGDQASISC RSSQNIVHSNGNTYLE WYLQK
PGQSPKLLIY KVSNRFS GVPDRFSGSGSGTDFTLKITRLEAEDLGV
YYC FQGSHVPWT FGGGTKLEIK 64 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH B-N10 QVQLKQSGPGLLQPSQSLSISCTVS GFSLATYGVH WVRQSPGKG
LEWLGVIWRG GSTDYSAAFMS RLSITKDNSKSQVFFKMNSLQAD
DTAIYFCAK QAYGHYMDY WGQGTSVTVSS 63 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 Elotuzumab Light chain VL Elotuzumab DIQMTQSPSSLSSASVGDRVTITCKAS QDVGIA VAWYQQKPGKVP
KLLIY WAS TRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYC QQ
YSSYPYT FGQGTKVEIK 50 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH Elotuzumab EVQLVESGGGLVQPGGSLRLSCAASGFDFS RYWMS WVRQAPGK
GLEWIGE INPDSST I NYAPSLKDKFIISRDNAKNSLYLQMNSLRAE
DTAVYYC ARPDGNYWYFDV WGQGTLVTVSS 49 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 G631mAb Light chain VL G631 DIQMTQSPSSLSSASVGDRVTITC RASQDVSTAVA WYQQKPGKAP
KLLIY SASFLYS GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQ
GYGNPFT FGQGTKVEIK 58 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH G631 EVQLVESGGGLVQPGGSLRLSCAASGFTIS DYWIH WVRQAPGKG
LEWVA GITPAGGYTYYADSVKG RFTISADTSKNTAYLQMNSLR
AEDTAVYYCAR FVFFLPYAMDY WGQGTLVTVSS 57 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 YervoymAb Light chain VL Yervoy EIVLTQSPGTLSLSPGERATLSC RASQSVGSSYLA WYQQKPGQAP
RLLIY GAFSRAT GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQ
YGSSPWT FGQGTKVEIK 28 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH Yervoy QVQLVESGGGVVQPGRSLRLSCAASGTFFS SYTMH WVRQAPGK
GLEWVTFI SYDGNNKYYADSVKG RFTISRDNSKNTLYLQMNSLR
AEDTAIYYCAR TGWLGPFDY WGQGTLVTVSS 27 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120 NM3E2mAb Light chain VL NM3E2 SELTQDPAVSVALGQTVRITC QGDSLRSYYAS WYQQKPGQAPVL
VIY GKNNRPS GIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSR
DSSGNHVV FGGGTKLTVL 48 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVD
NALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACE
VTHQGLSSPVTKSFNRGEC 104 Heavy chain VH NM3E2 EVQLVESGGGVVRPGGSLRLSCAASGFTF DDYGMS WVRQAPGK
GLEWVSG INWNGGSTGYADSVKG RFTISRDNAKNSLYLQMNSL
RAEDTAVYYCAR GRSLLFDY WGQGTLVTVSR 47 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGA
LTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN
TKVDKKVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEV
KFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN
GKEYKCKVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNG
QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH
EALHNHYTQKSLSLSPGK 120

The expression levels of some antibodies with structure 1 of the bispecific antibody of the present invention are shown in FIG. 5A-D. Fig. 5A shows the expression levels upon transient transfection of Y100-A series and Y100-B series antibodies in CHO cells. Fig. 5B shows the expression levels upon transient transfection of Y101/Y103/Y104/Y105-A series and Y101/Y103/Y104/Y105-B series antibodies in CHO cells. Fig. 5C shows the expression levels upon transient transfection of Y106/Y110/Y116-A series and Y106/Y110/Y116-B series antibodies in CHO cells. Fig. 5D shows the purity determined by size exclusion HPLC after protein A affinity chromatography for the Y100/Y101/Y103/Y104/Y105/Y106/Y110/Y116-A and B series antibodies that are expressed at levels greater than 5 mg/L.

Based on FIG. 5A-C, it can be seen that the expression level upon transient transfection of two series of bispecific antibodies of structure 1 (i.e. series A and series B) is significantly different, with the expression level of series A rarely exceeding 10 mg/l, and the expression level of series B above. Moreover, as can be seen from FIG. 5D, the purity of the structure 1 bispecific antibody of series B is more than 70%, and the purity of the structure 1 bispecific antibody of series A is substantially the same as that of series B. The purity of antibodies whose expression levels are less than 5 mg/L is not determined.

Fig. 6 shows the expression level of some antibodies with bispecific antibody structure 2 for comparison. In particular, FIG. 6 shows the expression levels of Y200/Y201/Y203/Y204/Y205/Y206/Y210/Y216-A series and B series antibodies transiently transfected into CHO cells, and the purity determined by size exclusion HPLC after Protein A affinity chromatography.

As can be seen from FIG. 6, two series of antibodies with bispecific antibody structure 2 (ie, series A and series B) do not have a significant difference in the transient expression level, and the expression level does not exceed 20 mg/L. Moreover, regarding the purity of bispecific antibody structure 2 determined by size exclusion HPLC after protein A affinity chromatography, the result shows that the purity of different molecules varies greatly from 30% to 90%. The expression level of some antibodies is too low to determine purity.

Fig. 7 shows the expression level of some antibodies with bispecific antibody structure 3 for comparison. In particular, FIG. 7 shows the expression levels of transient transfection of Y300/Y304/Y316 series A, series B, series C and series D antibodies in CHO cells and the purity determined by size exclusion HPLC after protein A affinity chromatography.

As can be seen from FIG. 7, the expression levels upon transient transfection of four series of bispecific antibodies of structure 3 (ie, series A, series B, series C, and series D) are not high, and the expression levels do not exceed 12 mg/L. Moreover, regarding the purity of antibodies with bispecific antibody structure 3 determined by size exclusion HPLC after Protein A affinity chromatography, the result shows that the purity is low and not more than 50%. The expression level of some antibodies is too low to determine purity.

Fig. 8 shows the expression level of some antibodies with bispecific antibody structure 4 for comparison. In particular, FIG. 8 shows the expression level of transient transfection of Y400/Y404/Y416 series A and series B antibodies in CHO cells and the purity determined by size exclusion HPLC after protein A affinity chromatography.

As can be seen from FIG. 8, the expression level of two series of bispecific antibodies of structure 4 (ie, series A and series B) is not high, and the expression level does not exceed 6 mg/L. Moreover, regarding the purity of the bispecific antibody of structure 4 determined by size exclusion HPLC after protein A affinity chromatography, the result shows that the purity varies greatly from 30% to 90%. The expression level of some antibodies is too low to determine purity.

The expression level and purity between antibodies of bispecific antibody structures 1-4 with the same target and antibody variable region sequence were compared and the results are shown in FIG. 9A-I. Fig. 9A shows the expression level and purity of antibodies with structure 1-4 of a bispecific antibody with the variable region sequences of the antibody S70 (SEQ ID NOs: 41 and 42) and the variable region sequences of the antibody G631 (SEQ ID NOs: 57 and 58). Fig. 9B shows the expression level and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequences of the antibody S70 (SEQ ID NOs: 41 and 42) and the variable region sequences of the antibody 3G12 (SEQ ID NOs: 59 and 60). Fig. 9C shows the expression level and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequences of antibody 12A4 (SEQ ID NOs: 45 and 46) and the variable region sequences of antibody 3G12 (SEQ ID NOs: 59 and 60). Fig. 9D shows the expression level and purity of bispecific antibody structure 1 and 2 antibodies with the variable region sequences of the antibody S70 (SEQ ID NOs: 41 and 42) and the variable region sequences of the antibody LAG35 (SEQ ID NOs: 33 and 34). Fig. 9E shows the expression level and purity of antibodies with structures 1, 3 and 4 of the bispecific antibody with the variable region sequences of the 5C4 antibody (SEQ ID NOs: 37 and 38) and the variable region sequences of the Yervoy antibody (SEQ ID NOs: 27 and 28). Fig. 9F shows the expression level and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequences of the antibody 5C4 (SEQ ID NOs: 37 and 38) and the variable region sequences of the antibody 10A7 (SEQ ID NOs: 29 and 30). Fig. 9G shows the expression level and purity of bispecific antibody structure 1 and 2 antibodies with the variable region sequences of antibody 5C4 (SEQ ID NOs: 37 and 38) and the variable region sequences of antibody G631 (SEQ ID NOs: 57 and 58). Fig. 9H shows the expression level and purity of antibodies with structure 1 and 2 of the bispecific antibody with the variable region sequences of the antibody 5C4 (SEQ ID NO: 37 and 38) and the variable region sequences of the antibody B-N10 (SEQ ID NO: 63 and 64). Fig. 9I shows the expression level and purity of antibodies with bispecific antibody structure 1-4 with Elotuzumab antibody variable region sequences (SEQ ID NOs: 49 and 50) and NM3E2 antibody variable region sequences (SEQ ID NOs: 47 and 48).

As can be seen from FIG. 9A, an antibody with bispecific antibody structure 1 has a higher level of expression and purity compared to other structures, with Y100-B6 and Y100-B7 having better expression and/or purity compared to antibodies of other structures, and Y100-B7 has the best level of expression and purity; Fig. 9B shows that antibodies with bispecific antibody structure 1 have higher expression level and purity compared to other structures, with Y101-B1 and Y101-B2 having better expression level and purity compared to antibodies of other structures, and Y101-B2 having the best level of expression and purity; Fig. 9C shows that antibodies with bispecific antibody structure 1 have a higher level of expression and/or antibody purity compared to other structures, with Y101-B3 and Y101-B4 having better expression level and purity than antibodies with bispecific antibody structure 2. and Y101-B4 has the best expression level and purity; Fig. 9D shows that bispecific antibody structure 1 antibodies have better expression level and purity compared to other structures, with Y103-B1 and Y103-B2 having better expression level and purity than bispecific antibody structure 2 antibodies, and Y103-B2 has the best level of expression and purity; Fig. 9E shows that antibodies with structure 1 bispecific antibodies have better expression level and/or antibody purity compared to other structures, with Y104-B1 and Y104-B2 having better expression level and purity than antibodies with structure 3 and structure 4 bispecific antibodies, and Y104-B2 has the best expression level and purity; Fig. 9F shows that bispecific antibody structure 1 antibodies have better expression level and/or antibody purity compared to other structures, with Y105-B1 and Y105-B2 having better expression level and purity than bispecific antibody structure 2 antibodies, and Y105-B2 has the best expression level and purity; as can be seen from Fig. 9G, antibodies with bispecific antibody structure 1 have better expression level and antibody purity compared to other structures, with Y106-B1 and Y106-B2 having better expression level and/or purity than bispecific antibody structure 2 antibodies, and Y106- B2 has the best expression level and purity; as can be seen from Fig. 9H, antibodies with bispecific antibody structure 1 have a higher level of expression and purity compared to other structures, with Y110-B1 and Y110-B2 having better expression and purity than antibodies with bispecific antibody structure 2, and Y110-B2 has the best level of expression and purity; Fig. 9I shows that antibodies with bispecific antibody structure 1 have higher expression level and/or antibody purity compared to other structures, with Y116-B1 and Y116-B2 having better expression level and purity compared to antibodies of other bispecific antibody structures. and Y116-B2 has the best expression level and purity.

If the hinge region, CH2 domain, and CH3 domain of each of the above antibodies were replaced with corresponding sequences from IgG2 (GenBank accession no. MH025834.2), IgG3 (GenBank accession no. AJ390278), or IgG4 (GenBank accession no. KJ901516), then the variable region sequence of the antibody and linker remained the same to produce the substituted antibody, there was no significant difference in expression level and purity between the resulting substituted antibody and the original antibody. Based on the data presented above, it can be seen that bispecific antibody structure 1 has significant advantages over other bispecific antibody structures in terms of expression level and purity.

In addition, antibodies such as Y100-B7, Y101-B2 and Y101-B4 are stably expressed in the pool of CHO cells with an expression level of more than 700 mg/L and a purity of more than 85%. The antibody was subjected to Protein A affinity chromatography followed by monoclonal antibody purification methods such as conventional anion/cation exchange chromatography to obtain the final product of interest with a purity of greater than 95% and an overall yield of 40%, consistent with data for monoclonal antibodies at their expression and purification.

Example 2. Detection of biological activity of bispecific antibodies

1. Affinity for antigen

1) Antigen preparation: for antigenic proteins VEGFA, TGF-β1, IL-10, PD-1, CTLA-4, TIGIT, SLAMF7, LAG-3 and CD16a (sequences, respectively, see Table 26), etc. . An expression plasmid pcDNA3.1 (purchased from invitrogen) with a His tag was constructed, where the extracellular domains of five proteins PD-1, CTLA-4, TIGIT, SLAMF7 and CD16a were selected for the construction, then transiently transfected into 293E cells for expression and cleaning; The purification method was divided into two stages, i.e. Nickel chelate column chromatography purification and molecular sieve purification resulting in antigenic protein purity determined by SDS-PAGE of at least 95%; the concentration of each antigenic protein was adjusted to 2 μg/ml, and the microtiter plate was coated with antigenic protein at 100 μl/well, 4°C overnight; the supernatant was discarded, and 250 μl of blocking solution (3% BSA in PBS) was added to each well;

2) addition of antibody: according to the experimental design, the procedure was carried out at room temperature and the antibody was diluted in a gradient with 1% BSA in PBS. For example, the initial dilution antibody concentration was 3000 nM, and the antibody was diluted 3-fold with 11 gradients. The diluted antibody was added to the wells of the microtiter plate at 200 μl/well, incubated at room temperature for 2 h and the supernatant was then discarded;

3) washing: the plate was washed with PBST (PBS containing 0.1% Tween20) at 200 μl/well 3 times;

4) Secondary antibody incubation: diluted secondary antibody HRP-labeled anti-human IgG (Sigma, A8792) was added in a volume of 100 μl/well and incubated at room temperature for 1 hour, where the secondary antibody was diluted at 1:40000 and the diluent was 1% BSA in PBS, and the wells were coated with antigen only and the secondary antibody was used as a control;

5) washing: the plate was washed with PBST (PBS containing 0.1% Tween20) at 200µl/well 5 times;

6) color development: TMB color development solution (obtained from color development solutions A and B purchased from Wuhan Boster Company, and mixed according to A:B=1:1, ready for use) was added at 100μL/well, and color development was carried out at 37°C for 5 minutes;

7) 2M HCl stop solution was added at 100µL/well, and then it was mandatory to use a microplate reader to read at 450nm for 30 minutes, and the data were presented graphically using Graphpad Prism 5 software, with antibody concentration (nM) according to the abscissa axis and the average fluorescence intensity along the ordinate axis; The EC50 value, which indicates the affinity of the antibody for the corresponding target antigen, was calculated by the single-site specific binding method. The results are shown in Table 32. The results show that in the case of antibodies with identical variable region sequences, the affinity of the bispecific antibody of the present invention (i.e., bispecific antibody of structure 1) for the corresponding target antigen is significantly higher than that of bispecific antibodies of structures 2- 4. In addition, the bispecific antibodies of the present invention (bispecific antibodies of structure 1) have the strongest antigen binding activity against the target.

2. Cellular affinity

1) Cell preparation: PD-L1 positive H358 cells (purchased from Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) were used to determine the binding activity of bispecific antibody molecules against PD-L1 cells at the terminal stage of differentiation; a sufficient number of cells were centrifuged at 300×g for 5 minutes, the supernatant was discarded; cells were resuspended in 1% FBS-PBS, adjusted to a density of 4×10 ⁶ /ml; 50 μl of cells were selected for each well and seeded at 2×10 ⁵ per well; in this case, centrifugation was carried out at 300×g at 4°C for 5 minutes, the supernatant was drained and cell seeding was carried out on ice;

2) Addition of antibody: according to the experimental design, antibodies were diluted in a gradient and antibody dilution was carried out on ice; for example, the initial antibody concentration for dilution was 3000 nM, and the antibody was diluted 3-fold with 11 concentration gradients; the diluted antibody was added to the well at 50 μl per well, mixed thoroughly with gentle purging and incubated at 4°C with shaking at 1100 rpm for 2 hours;

3) washing: cells were resuspended with 150 μl of 1% FBS-PBS and centrifuged at 300 × g at 4°C for 5 minutes; the supernatant was discarded; and the cells were washed one more time;

4) secondary antibody incubation: diluted secondary antibody PE anti-human IgG FC (Biolegend, 409304) was added in a volume of 50 µl/well, the final concentration of the secondary antibody was 8 µg/ml, mixed thoroughly with gentle purging and incubated at 4°C in the dark with shaking at 1100 rpm for 1 hour; while the well to which only cells and secondary antibody were added was used as a control;

5) washing: cells were resuspended with 150 μl of 1% FBS-PBS and centrifuged at 300 g at 4°C for 5 minutes, the supernatant was discarded; the cells were washed one more time;

6) fixation: 2% paraformaldehyde solution was added at 200μL/well to resuspend and fix the cells at room temperature for 20 minutes, and then the cells were centrifuged at 300×g for 5 minutes; the supernatant was discarded;

7) cell resuspension: cells were resuspended with 200 μl of 1% FBS-PBS, centrifuged at 300 × g for 5 minutes, and the supernatant was discarded;

8) loading for flow cytometry: cells were resuspended with 150 μl of 1% FBS-PBS and detection was carried out on a flow cytometer;

9) Data analysis: Data were analyzed using flow analysis software FlowJo 7.6 to obtain the average fluorescence intensity at a specific antibody concentration, and presented graphically using Graphpad Prism 5 software, with antibody concentration (nM) on the x-axis and average fluorescence intensity on the x-axis. ordinate; the single site specific binding method was used to calculate the EC50 value, which indicates the cellular affinity of the antibody for the corresponding target antigens; the results are shown in Table 32.

Table 32
Binding activity of some antibodies having bispecific antibody structure 1-4 Antibody code H358 cell binding activity EC50(pM) Binding activity to VEGF antigen EC50(pM) Binding activity to TGF-β1 antigen EC50 (pM) Binding activity to LAG-3 antigen EC50 (pM) PD-1 antigen binding activity EC50 (pM) Binding activity to CTLA-4 antigen EC50
(pM) Binding activity to TIGIT antigen EC50 (pM) Binding activity to IL-10 antigen EC50 (pM) Binding activity to SLAMF7 antigen EC50 (pM) Binding activity to CD16a antigen EC50 (pM) Y100-B6
Y100-B7
Y200-A1
Y200-B1
Y200-B3
Y300-A1
Y400-B1 170.60
169.60
156.24
173.00
207.15
180.95
254.02 34.09
35.19
237.93
188.41
47.33
307.96
268.35 N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A.
N.A. Y101-B1
Y101-B2
Y201-A1
Y201-B1 179.42
191.23
173.80
192.18 N.A.
N.A.
N.A.
N.A. 5.97
3.09
25.05
28.46 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y101-B3
Y101-B4
Y201-A2
Y201-B2 169.40
119.07
143.63
124.05 N.A.
N.A.
N.A.
N.A. 5.46
1.86
31.18
28.01 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y103-B1
Y103-B2
Y203-A1
Y203-B1 181.84
169.09
143.09
187.96 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. 16.61
12.68
65.70
89.44 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y104-B1
Y104-B2
Y304-A1
Y404-B1 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. 127.65
93.78
464.65
388.51 56.24
69.68
468.46
330.93 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y105-B1
Y105-B2
Y205-A2
Y205-B2 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. 114.13
114.70
306.23
155.11 N.A.
N.A.
N.A.
N.A. 90.17
86.97
484.29
409.16 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y106-B1
Y106-B2
Y206-A2
Y206-B1
Y206-B2 N.A.
N.A.
N.A.
N.A.
N.A. 66.33
53.91
63.03
212.78
59.64 N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A. 102.46
112.44
449.79
164.52
311.52 N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A. Y110-B1
Y110-B2
Y210-A1
Y210-B1 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. 129.81
113.27
148.67
198.43 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. 58.79
66.62
409.05
234.03 N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A. Y116-B1
Y116-B2
Y216-A1
Y216-B1
Y316-A1
Y416-B1 N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. N.A.
N.A.
N.A.
N.A.
N.A.
N.A. 636.49
648.23
714.64
499.21
9176.07
5590.07 564.05
436.73
3257.47
3220.97
3842.87
3557.75 S70mAb 175.64 N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. 12A4mAb 125.69 N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. G631mAb N.A. 30.00 N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. 3G12mAb N.A. N.A. 2.60 N.A. N.A. N.A. N.A. N.A. N.A. N.A. LAG35mAb N.A. N.A. N.A. 10.41 N.A. N.A. N.A. N.A. N.A. N.A. 5C4mAb N.A. N.A. N.A. N.A. 111.42 N.A. N.A. N.A. N.A. N.A. Yervoy mAb N.A. N.A. N.A. N.A. N.A. 59.27 N.A. N.A. N.A. N.A. 10A7mAb N.A. N.A. N.A. N.A. N.A. N.A. 83.14 N.A. N.A. N.A. BN10mAb N.A. N.A. N.A. N.A. N.A. N.A. N.A. 62.96 N.A. N.A. Elotuzumab N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. 628.00 N.A. NM3E2mAb N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. N.A. 520.10

From the data in Table 32, it can be seen that when the sequences of the variable regions of different antibodies are identical, the binding activity of such antibodies having different bispecific antibody structures with the target antigen differs significantly, while for any targets, antibodies with structure 1 of the bispecific antibody have the strongest antigen binding activity . If the hinge region, CH2 domain and CH3 domain of each of the above antibodies are replaced with the corresponding sequences from IgG2, IgG3 or IgG4 (supra), and the antibody variable region and linker sequences remain the same, resulting in a substituted antibody, there is no significant difference in antigen binding activity between the resulting substituted antibody and the original antibody.

Example 3 Antibody Stability Test

Experimental procedure:

1. The specific steps of the accelerated thermal stability test at 40°C are as follows:

1) the sample was transferred to a specific buffer, the buffer component is 20 mM citric acid, pH 5.5, and the sample concentration was adjusted to 1 mg/ml;

2) each sample was divided into 500 μl per tube (6 tubes in total) and the tubes were hermetically sealed and placed in a 40°C water bath. On day 0, day 3, day 5, day 7, day 10 and day 14, samples were taken for size exclusion HPLC. The water bath was maintained for a total of 14 days.

2. The acid fastness test, also known as the low pH stability test, is a test to determine whether an antibody molecule can retain its original state after being treated under acidic conditions for a specified period of time, followed by neutralization to physiological conditions. The specific stages are as follows:

when antibody molecules were subjected to protein A affinity chromatography, the eluted antibody solution was not neutralized in the acid elution step (citrate buffer was used at pH 3.5); After storage in the specified buffer for a certain period of time, samples were taken at 30 minutes and 60 minutes, 1/10 volume of 1M Tris-HCl (pH 8.0) was added to neutralize and the samples were subjected to size exclusion HPLC detection.

3.1 Stability test for bispecific antibody with structure 1

(1) Accelerated thermal stability testing at 40°C of bispecific antibodies with different structures

The test results are shown in Fig. 10A-E. Fig. 10A shows the purity of Y100-B6, Y100-B7, Y200-A1, Y200-B1, Y200-B3, Y300-A1 and Y400-B1 determined by size exclusion HPLC on day 0, day 7 and day 14 after they were processed at 40°C; Fig. 10B shows the purity of Y101-B1, Y101-B2, Y201-A1, Y201-B1, Y101-B3, Y101-B4, Y201-A2 and Y201-B2 determined by size exclusion HPLC on day 0, day 7 and day 14, after how they were processed at 40°C; Fig. 10C shows the purity of Y103-B1, Y103-B2, Y203-A1, Y203-B1, Y104-B1, Y104-B2, Y304-A1 and Y404-B1 determined by size exclusion HPLC on day 0, day 7 and day 14, after how they were processed at 40°C; Fig. 10D shows the purities of Y105-B1, Y105-B2, Y205-A2, Y205-B2, Y106-B1, Y106-B2, Y206-A2, Y206-B1 and Y206-B2 determined by size exclusion HPLC on day 0, day 7 and day 14, after they were treated at 40°C; Fig. 10E shows the purity of Y110-B1, Y110-B2, Y210-A1, Y210-B1, Y116-B1, Y116-B2, Y216-A1, Y216-B1, Y316-A1, Y416-B1 determined by size exclusion HPLC on day 0 , day 7 and day 14, after they were treated at 40°C.

As can be seen from FIG. 10, the purity of various antibodies with structure 1 bispecific antibody was maintained at more than 90% after treatment at 40°C for 14 days, without much aggregation or degradation, and this indicates that the bispecific antibody structure 1 has good thermal stability. The purity of bispecific antibodies with other structures is significantly reduced after treatment at 40°C for 14 days.

(2) Acid resistance test of various bispecific antibody structures

The test results are shown in Fig. 11A-E. Fig. 11A shows the purity of Y100-B6, Y100-B7, Y200-A1, Y200-B1, Y200-B3, Y300-A1 and Y400-B1 determined by size exclusion HPLC after treatment under low pH conditions for 0 minutes, 30 minutes and 60 minutes; Fig. 11B shows the purity of Y101-B1, Y101-B2, Y201-A1, Y201-B1, Y101-B3, Y101-B4, Y201-A2 and Y201-B2 determined by size exclusion HPLC after treatment under low pH conditions for 0 minutes, 30 minutes and 60 minutes; Fig. 11C shows the purity of Y103-B1, Y103-B2, Y203-A1, Y203-B1, Y104-B1, Y104-B2, Y304-A1 and Y404-B1 determined by size exclusion HPLC after treatment under low pH conditions for 0 minutes, 30 minutes and 60 minutes; Fig. 11D shows the purity of Y105-B1, Y105-B2, Y205-A2, Y205-B2, Y106-B1, Y106-B2, Y206-A2, Y206-B1 and Y206-B2 determined by size exclusion HPLC after treatment under low pH conditions in for 0 minutes, 30 minutes and 60 minutes; Fig. 11E shows the purity of Y110-B1, Y110-B2, Y210-A1, Y210-B1, Y116-B1, Y116-B2, Y216-A1, Y216-B1, Y316-A1 and Y416-B1 determined by size exclusion HPLC after treatment in low pH conditions for 0 minutes, 30 minutes and 60 minutes.

As can be seen from FIG. 11A-E, when bispecific antibody structure 1 antibodies were treated for 60 minutes under low pH conditions, the purity of the antibodies did not change significantly, indicating that bispecific antibody structure 1 antibodies have good acid resistance. For other bispecific antibody structures, the purity of some antibodies decreased significantly after 60 min of low pH treatment.

Based on FIG. 10 and 11, it can be seen that the antibody with bispecific antibody structure 1 has better heat stability and acid resistance compared with other bispecific antibody structures.

When the hinge region, CH2 domain and CH3 domain of each of the above antibodies are replaced with the corresponding sequence from IgG2, IgG3 or IgG4 (supra), and the antibody variable region and linker sequences remain the same, resulting in a substituted antibody, there is no significant difference in stability between the resulting the substituted antibody and the original antibody.

Example 4. Experiment 1 to determine the pharmacodynamics of bispecific antibodies in vivo

1). Materials used for the experiment:

Cells: MC38 (mouse colon cancer cell line purchased from ATCC);

Mice: C57BL/6 mice, female, purchased from Beijing Vital River;

Inoculation method: MC38 cells were cultured and collected, the cell concentration was adjusted, then the cells were subcutaneously inoculated into the dorsal region at 1*10 ⁶ cells/mouse (0.1 ml/mouse). When the tumor grew to 100-200mm ³ , the mice were divided into injection groups, 8 mice in each group:

Test drug: Y100-B7;

Negative control: saline;

Positive control: Tecentriq (anti-PD-L1 monoclonal antibody, Roche);

Route of administration: Different doses (13.3mg/kg and 4mg/kg) were established for Y100-B7, and the administered dose of Tecentriq was 10mg/kg; the drug was administered intraperitoneally, starting on day 0, 3 times a week for one week.

Tumor volume: tumor length and width were measured every 2-3 days. When the tumor volume of a group approached 2000 mm ³ or the tumor volume of an individual mouse reached 3000 mm ³ , the group stopped the trial.

2). Experiment results

Pharmacodynamics test results in vivo for Y100-B7 are shown in Fig. 12. Fig. 12 shows the in vivo pharmacodynamics of bispecific antibody Y100-B7 in a mouse tumor model and tumor volume. The results show that the group receiving the high dose of Y100-B7 (13.3 mg/kg) showed significant antitumor effect, and this effect was better than Tecentriq.

Example 5. Experiment 2 to determine the pharmacodynamics of bispecific antibodies in vivo

1). Materials used for the experiment:

Cells: MC38 (mouse colon cancer cell line purchased from ATCC);

Mice: C57BL/6 mice, female, purchased from Beijing Vital River;

Inoculation method: MC38 cells were cultured, collected, cell concentration was adjusted, then the cells were subcutaneously inoculated into the dorsal region at 1*10 ⁶ cells/mouse (0.1 ml/mouse). When the tumor grew to 100-200mm ³ , the mice were divided into injection groups, 8 mice in each group:

Test drug: Y101-B2;

Negative control: saline;

Positive control: Tecentriq (anti-PD-L1 monoclonal antibody, Roche);

Route of administration: Different doses (13.3mg/kg and 4mg/kg) were established for Y101-B2, and the administered dose of Tecentriq was 10mg/kg; the drug was administered intraperitoneally, starting on day 0, 3 times a week for 3 weeks.

Tumor volume: tumor length and width were measured 3 times a week. When the tumor volume of a group approached 2000 mm ³ or the tumor volume of an individual mouse reached 3000 mm ³ , the group stopped the trial.

2). Experiment results

Pharmacodynamics test results in vivo for Y101-B2 are shown in Fig. 13. Fig. 13 shows the in vivo pharmacodynamics of bispecific antibody Y101-B2 in a mouse tumor model and tumor volume. The results show that the group receiving the high dose of Y101-B2 (13.3 mg/kg) showed significant antitumor effect, and this effect was better than Tecentriq.

Example 6: Preparation and determination of biological activity of biparatope antibody

A biparatope antibody is a type of bispecific antibody and can bind to two different epitopes of the same antigen.

The expression plasmid construction, transient transfection of 293E cells, and antibody production method were the same as the methods described above.

The antigen in question was the extracellular domain of human Her2 (sequence source: UniProtKB-P04626).

The corresponding antibody variable region sequences are shown in the table below:

Table 33
Anti-Her2 antibody variable region sequences Antibody code
(sequence source) Amino acid sequences of anti-Her2 antibody variable regions (bold and underlined amino acids represent CDR regions) VH SEQ ID NO. VL SEQ ID NO. Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH
WVRQAPGKGLEWVA RIYPTNGYTRYADSVKG RFT
ISADTSKNTAYLQMNSLRAEDTAVYYCSR WGGDG
FYAMDY WGQGTLVTVSS 146 DIQMTQSPSSLSSASVGDRVTITC RASQDVNT
AVA WYQQKPGKAPKLLIY SASFLYS GVPSRF
SGSRSGTDFTLTISSLQPEDFATYYC QQHYTT
PPT FGQGTKVEIK 147 Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD
WVRQAPGKGLEWVA DVNPNSGGSIYNQRFKG RFT
LSVDRSKNTLYLQMNSLRAEDTAVYYCAR NLGPSF
YFDY WGQGTLVTVSS 148 DIQMTQSPSSLSSASVGDRVTTITC KASQDVSIG
VA WYQQKPGKAPKLLIY SASYRYT GVPSRFS
GSGSGTDFTLTISSLQPEDFATYYC QQYYIYP
YT FGQGTKVEIK 149

The sequences of the constructed antibodies are shown in the following tables:

Table 34
Antibody codes and amino acid sequences of some antibodies with the structure of 1 bispecific antibody Antibody code Peptide Domain Code Amino acid sequences (bold and underlined amino acids represent CDR regions) SEQ ID NO. Herceptin mAb Heavy chain VH Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLEW
VA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTAVY
YCSR WGGDGFYAMDY WGQGTLVTVSS 146 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Light chain VL Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Perjeta mAb Heavy chain VH Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Light chain VL Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Y140-A1 Hybrid light chain VLa Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VLb Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 Hybrid heavy chain VHa Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VHb Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFN
WYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKC
KVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-A2 Hybrid light chain VLa Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VLb Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 Hybrid heavy chain VHa Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VHb Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFN
WYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKC
KVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-A3 Hybrid light chain VLa Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VLb Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 Hybrid heavy chain VHa Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VHb Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-A4 Hybrid light chain VLa Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VLb Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPPT FGQGTKVEIK 147 Hybrid heavy chain VHa Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VHb Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-B1 Hybrid light chain VLa Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VHb Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 Hybrid heavy chain VHa Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VLb Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFN
WYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKC
KVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-B2 Hybrid light chain VLa Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VHb Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 Hybrid heavy chain VHa Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VLb Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G2DCH2 PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEAPEVQFN
WYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKC
KVSNKGLPAPIEKTISKTK 117 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-B3 Hybrid light chain VLa Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VHb Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 Hybrid heavy chain VHa Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VLb Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120 Y140-B4 Hybrid light chain VLa Perjeta DIQMTQSPSSSLSASVGDRVTITC KASQDVSIGVA WYQQKPGKAPKLLI
Y SASYRYT GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC QQYYIYPYT
FGQGTKVEIK 149 C.L. Lc1 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNAL
QSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC 104 Linker1 Lin17 GGGGSEAAAKGGGGS 85 VHb Herceptin EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTYIH WVRQAPGKGLE
WVA RIYPTNGYTRYADSVKG RFTISADTSKNTAYLQMNSLRAEDTA
VYYCSR WGGDGFYAMDY WGQGTLVTVSS 146 Hybrid heavy chain VHa Perjeta EVQLVESGGGLVQPGGSLRLSCAAS GTFTDYTMD WVRQAPGKGLE
WVA DVNPNSGGSIYNQRFKG RFTLSVDRSKNTLYLQMNSLRAEDTA
VYYCAR NLGPSFYFDY WGQGTLVTVSS 148 CH1 G1CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS
GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDK
KVEPKSC 111 Linker2 Lin17 GGGGSEAAAKGGGGS 85 VLb Herceptin DIQMTQSPSSLSSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLL
IY SASFLYS GVPSRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPP
T FGQGTKVEIK 147 Linker3 Lin14 GGGGSGGGGSGGGGS 82 Hinge Hin1 DKTHTCP 91 CH2 G1CH2 PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAK 115 CH3 G1CH3 GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE
NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH
YTQKSLSLSPGK 120

The expression level upon transient transfection of both Herceptin mAb and Perjeta mAb is 30 mg/L; the expression level during transient transfection of the bispecific antibody Y140-B1/B2/B3/B4 is not less than 25 mg/l; the expression level of the bispecific antibody Y140-A1/A2/A3/A4 is 7 mg/l - 20 mg/l, and the purity of all antibodies according to size exclusion chromatography is no less than 80%.

2. Detection of biological activity

Cell affinity

1) Cell preparation: Her2-positive BT-474 cells (purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences) were used to determine the affinity of the bispecific antibody molecule for the Her2 end; a sufficient number of cells were centrifuged at 300×g for 5 minutes, the supernatant was discarded; cells were resuspended in 1% FBS-PBS, adjusted to a density of 2x10 ⁶ /ml, and 50 μl of cells per well were collected and then seeded at 1x10 ⁵ per well; in this case, the cells were centrifuged at 300×g at 4°C for 5 minutes, the supernatant was drained, and the cells were seeded on ice;

2) Antibody addition: According to the experimental design, the antibody shown in Table 34 was diluted in a gradient and the antibody dilution was carried out on ice; for example, the initial antibody concentration for dilution was 3000 nM, and the antibody was diluted 3-fold with 11 concentration gradients; the diluted antibody was added to the well at 50 μl per well, mixed thoroughly with gentle purging and incubated at 4°C with shaking at 1100 rpm for 2 hours;

3) washing: cells were resuspended with 150 μl of 1% FBS-PBS and centrifuged at 300 × g at 4°C for 5 minutes; the supernatant was discarded; and the cells were washed one more time;

4) secondary antibody incubation: diluted secondary antibody PE anti-human IgG FC (Biolegend, 409304) was added at a volume of 50 µl/well, the final concentration of the secondary antibody was 8 µg/ml, mixed thoroughly with gentle purging and incubated at 4°C in the dark with shaking at 1100 rpm for 1 hour; while the well to which only cells and secondary antibody were added was used as a control;

5) washing: cells were resuspended with 150 μl of 1% FBS-PBS and centrifuged at 300 g at 4°C for 5 minutes, the supernatant was discarded; the cells were washed one more time;

6) fixation: 2% paraformaldehyde solution was added at 200μL/well to resuspend and fix the cells at room temperature for 20 minutes, and then the cells were centrifuged at 300×g for 5 minutes; the supernatant was discarded;

7) cell resuspension: cells were resuspended with 200 μl of 1% FBS-PBS, centrifuged at 300 × g for 5 minutes, and the supernatant was discarded;

8) loading for flow cytometry: cells were resuspended with 150 μl of 1% FBS-PBS and detection was carried out on a flow cytometer;

9) Data analysis: Data were analyzed using flow analysis software FlowJo 7.6 to obtain the average fluorescence intensity at a specific antibody concentration, and presented graphically using Graphpad Prism 5 software, with antibody concentration (nM) on the x-axis and average fluorescence intensity on the x-axis. ordinate; the single-site specific binding method was used to calculate the EC50 value, which indicates the cellular affinity of the antibody for the corresponding target antigens.

Table 35
Binding activity of some bispecific antibodies Antibody code Binding activity to BT-474 EC50 cells (pM) Herceptin mAb 2567.33 Perjeta mAb 5456.55 Y140-A1 1931.84 Y140-A2 1313.78 Y140-A3 1889.42 Y140-A4 1537.68 Y140-B1 838.577 Y140-B2 813.730 Y140-B3 526.554 Y140-B4 420.308

From the above table, it can be seen that the affinity of bispecific antibodies formed by two antibodies that target the same antigen but different epitopes is significantly improved compared to the affinity of two monoclonal antibodies, and the affinity of bispecific B series antibodies is significantly stronger than for series A antibodies.

--->

LIST OF SEQUENCES

<110> WUHAN ISA BIOPHARMA CO., LTD.

<120> QUAD VALENT SYMMETRICAL BISPECIFIC ANTIBODIES

<130>FP200576MX

<160> 149

<170> Patent In version 3.3

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Gln Val Gln Leu Val Glu 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 Thr Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp

50 55 60

Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Phe

100 105 110

Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 2

<211> 109

<212> Protein

<213> Artificial sequence

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Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly

1 5 10 15

Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser

20 25 30

Asn Tyr Ala Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly

35 40 45

Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Val Pro Ala Arg Phe

50 55 60

Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val

65 70 75 80

Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn

85 90 95

Leu Trp Val Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 3

<211> 125

<212> Protein

<213> Artificial sequence

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Gln Val Gln Leu Val Glu 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 Thr Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp

50 55 60

Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Ala Arg His Gly Asn Phe Gly Asn Ser Tyr Val Ser Trp Ala

100 105 110

Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 4

<211> 109

<212> Protein

<213> Artificial sequence

<400> 4

Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly

1 5 10 15

Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser

20 25 30

Asn Tyr Ala Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly

35 40 45

Leu Ile Gly Gly Thr Asn Lys Arg Ala Pro Gly Val Pro Ala Arg Phe

50 55 60

Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val

65 70 75 80

Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Ala Leu Trp Tyr Ser Asn

85 90 95

Leu Trp Val Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

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<212> Protein

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Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asn Lys Tyr

20 25 30

Ala Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Arg Ile Arg Ser Lys Tyr Asn Asn Tyr Ala Thr Tyr Tyr Ala Asp

50 55 60

Ser Val Lys Asp Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr

65 70 75 80

Ala Tyr Leu Gln Met Asn Asn Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Val Arg His Gly Asn Phe Gly Asn Ser Tyr Ile Ser Tyr Trp

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Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

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<210> 6

<211> 109

<212> Protein

<213> Artificial sequence

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Gln Thr Val Val Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly

1 5 10 15

Thr Val Thr Leu Thr Cys Gly Ser Ser Thr Gly Ala Val Thr Ser Gly

20 25 30

Asn Tyr Pro Asn Trp Val Gln Gln Lys Pro Gly Gln Ala Pro Arg Gly

35 40 45

Leu Ile Gly Gly Thr Lys Phe Leu Ala Pro Gly Thr Pro Ala Arg Phe

50 55 60

Ser Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val

65 70 75 80

Gln Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Val Leu Trp Tyr Ser Asn

85 90 95

Arg Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 7

<211> 118

<212> Protein

<213> Artificial sequence

<400> 7

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Asp Ile Asn Trp Val Arg Gln Arg Pro Glu Gln Gly Leu Glu Trp Ile

35 40 45

Gly Trp Ile Phe Pro Gly Asp Gly Ser Thr Gln Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Thr Asp Thr Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

85 90 95

Ala Arg Gln Thr Thr Ala Thr Trp Phe Ala Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 8

<211> 107

<212> Protein

<213> Artificial sequence

<400> 8

Asp Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Thr Pro Gly

1 5 10 15

Asp Arg Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asp Tyr

20 25 30

Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile

35 40 45

Lys Tyr Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Ser Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Pro

65 70 75 80

Glu Asp Val Gly Val Tyr Tyr Cys Gln Asn Gly His Ser Phe Pro Leu

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys

100 105

<210> 9

<211> 120

<212> Protein

<213> Artificial sequence

<400> 9

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Ala Arg Pro Gly Ala

1 5 10 15

Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Trp Met Gln Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Thr Ile Tyr Pro Gly Asp Gly Asp Thr Arg Tyr Thr Gln Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Gln Leu Ser Ser Leu Ala Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Ile Pro Arg Leu Trp Tyr Phe Asp Val Trp Gly Ala

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 10

<211> 107

<212> Protein

<213> Artificial sequence

<400> 10

Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Asp Asn Leu Glu Gln

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 11

<211> 122

<212> Protein

<213> Artificial sequence

<400> 11

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 Val Ser Gly Phe Thr Phe Asn Ser Phe

20 25 30

Ala 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 Gly 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 Phe Cys

85 90 95

Ala Lys Asp Lys Ile Leu Trp Phe Gly Glu Pro Val Phe Asp Tyr Trp

100 105 110

Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 12

<211> 107

<212> Protein

<213> Artificial sequence

<400> 12

Glu Ile Val Leu Thr Gln Ser Pro Ala 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 Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 13

<211> 120

<212> Protein

<213> Artificial sequence

<400> 13

Gln 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 Ser Tyr

20 25 30

Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Ser Gly Asp Pro 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 Asp Leu Pro Leu Val Tyr Thr Gly Phe Ala Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 14

<211> 109

<212> Protein

<213> Artificial sequence

<400> 14

Asp Ile Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln

1 5 10 15

Thr Ala Arg Ile Ser Cys Ser Gly Asp Asn Leu Arg His Tyr Tyr Val

20 25 30

Tyr Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr

35 40 45

Gly Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr Gln Ala Glu

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Thr Tyr Thr Gly Gly Ala Ser Leu

85 90 95

Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln

100 105

<210> 15

<211> 120

<212> Protein

<213> Artificial sequence

<400> 15

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Phe Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Val Ile Pro Phe Leu Gly Ile Ala Asn Ser Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Asp Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Asp Ile Ala Ala Leu Gly Pro Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 16

<211> 107

<212> Protein

<213> Artificial sequence

<400> 16

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 Gly Ile Ser Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln 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 Asn Ser Tyr Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 17

<211> 115

<212> Protein

<213> Artificial sequence

<400> 17

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Leu Leu Trp Asn Tyr Trp Gly Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 18

<211> 109

<212> Protein

<213> Artificial sequence

<400> 18

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Ile Ile

35 40 45

Tyr Gly Ala Ser Thr Thr Ala Ser Gly Ile Pro Ala Arg Phe Ser Ala

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Pro

85 90 95

Ala Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 19

<211> 113

<212> Protein

<213> Artificial sequence

<400> 19

Glu Val Gln Leu Val Glu Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Ser Met His Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met

35 40 45

Gly Trp Ile Asn Thr Glu Thr Gly Glu Pro Thr Tyr Ala Asp Asp Phe

50 55 60

Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Thr Ala Val Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser

100 105 110

Ser

<210> 20

<211> 107

<212> Protein

<213> Artificial sequence

<400> 20

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Glu Arg Val Ser Leu Thr Cys Arg Ala Ser Gln Glu Ile Ser Val Ser

20 25 30

Leu Ser Trp Leu Gln Gln Glu Pro Asp Gly Thr Ile Lys Arg Leu Ile

35 40 45

Tyr Ala Thr Ser Thr Leu Asp Ser Gly Val Pro Lys Arg Phe Ser Gly

50 55 60

Ser Arg Ser Gly Ser Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Ser

65 70 75 80

Glu Asp Phe Val Asp Tyr Tyr Cys Leu Gln Tyr Ala Ser Tyr Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105

<210> 21

<211> 119

<212> Protein

<213> Artificial sequence

<400> 21

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Pro Asp Tyr

20 25 30

Tyr Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Tyr Phe Ala Ser Gly Asn Ser Glu Tyr Asn Gln Lys Phe

50 55 60

Thr Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Ser Leu Tyr Asp Tyr Asp Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Met Val Thr Val Ser Ser

115

<210> 22

<211> 112

<212> Protein

<213> Artificial sequence

<400> 22

Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Gln Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Ile Tyr Tyr Cys Ser Gln Ser

85 90 95

Ser Ile Tyr Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 23

<211> 121

<212> Protein

<213> Artificial sequence

<400> 23

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Arg Ile Ile Pro Ile Leu Gly Ile Ala Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Gly Tyr Tyr Ser His Asp Met Trp Ser Glu Asp Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 24

<211> 112

<212> Protein

<213> Artificial sequence

<400> 24

Leu Pro Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln

1 5 10 15

Arg Val Thr Ile Ser Cys Ser Gly Arg Ser Ser Asn Ile Gly Ser Asn

20 25 30

Ser Val Asn Trp Tyr Arg Gln Leu Pro Gly Ala Ala Pro Lys Leu Leu

35 40 45

Ile Tyr Ser Asn Asn Gln Arg Pro Pro Gly Val Pro Val Arg Phe Ser

50 55 60

Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln

65 70 75 80

Ser Glu Asp Glu Ala Thr Tyr Tyr Cys Ala Thr Trp Asp Asp Asn Leu

85 90 95

Asn Val His Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly

100 105 110

<210> 25

<211> 121

<212> Protein

<213> Artificial sequence

<400> 25

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Gly

20 25 30

Ser Tyr Phe Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ile Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg His Asp Gly Ala Val Ala Gly Leu Phe Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 26

<211> 108

<212> Protein

<213> Artificial sequence

<400> 26

Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Gln

1 5 10 15

Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Lys Ser Val

20 25 30

His Trp Tyr Gln Gln Pro Pro Gly Gln Ala Pro Val Val Val Val Tyr

35 40 45

Asp Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly Asn

50 55 60

Ser Asn Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu Ala Gly

65 70 75 80

Asp Glu Ala Val Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His

85 90 95

Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 27

<211> 118

<212> Protein

<213> Artificial sequence

<400> 27

Gln Val Gln Leu Val Glu 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

Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Thr Phe Ile Ser Tyr Asp Gly Asn Asn Lys 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 Ile Tyr Tyr Cys

85 90 95

Ala Arg Thr Gly Trp Leu Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

<210> 28

<211> 108

<212> Protein

<213> Artificial sequence

<400> 28

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 Gly 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 Phe 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 Tyr Gly Ser Ser Pro

85 90 95

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 29

<211> 119

<212> Protein

<213> Artificial sequence

<400> 29

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Thr Gln Pro Gly Lys

1 5 10 15

Ser Leu Lys Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Ser Ser Phe

20 25 30

Thr Met His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Phe Ile Arg Ser Gly Ser Gly Ile Val Phe Tyr Ala Asp Ala Val

50 55 60

Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Leu Leu Phe

65 70 75 80

Leu Gln Met Asn Asp Leu Lys Ser Glu Asp Thr Ala Met Tyr Tyr Cys

85 90 95

Ala Arg Arg Pro Leu Gly His Asn Thr Phe Asp Ser Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 30

<211> 113

<212> Protein

<213> Artificial sequence

<400> 30

Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Met Thr Cys Lys Ser Ser Gln Ser Leu Tyr Tyr Ser

20 25 30

Gly Val Lys Glu Asn Leu Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Ser Pro Lys Leu Leu Ile Tyr Tyr Ala Ser Ile Arg Phe Thr Gly Val

50 55 60

Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr

65 70 75 80

Ile Thr Ser Val Gln Ala Glu Asp Met Gly Gln Tyr Phe Cys Gln Gln

85 90 95

Gly Ile Asn Asn Pro Leu Thr Phe Gly Asp Gly Thr Lys Leu Glu Ile

100 105 110

Lys

<210> 31

<211> 123

<212> Protein

<213> Artificial sequence

<400> 31

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln

1 5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Glu Ser Gly

20 25 30

Leu Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

35 40 45

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

50 55 60

Leu Lys Ser Arg Ala Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

85 90 95

Cys Ala Arg Asp Gly Val Leu Ala Leu Asn Lys Arg Ser Phe Asp Ile

100 105 110

Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser

115 120

<210> 32

<211> 109

<212> Protein

<213> Artificial sequence

<400> 32

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 His Thr Val Arg Pro

85 90 95

Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105

<210> 33

<211> 120

<212> Protein

<213> Artificial sequence

<400> 33

Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu

1 5 10 15

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Asp Tyr

20 25 30

Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn His Arg Gly Ser Thr Asn Ser Asn Pro Ser Leu Lys

50 55 60

Ser Arg Val Thr Leu Ser Leu Asp Thr Ser Lys Asn Gln Phe Ser Leu

65 70 75 80

Lys Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala

85 90 95

Phe Gly Tyr Ser Asp Tyr Glu Tyr Asn Trp Phe Asp Pro Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 34

<211> 107

<212> Protein

<213> Artificial sequence

<400> 34

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Asn Leu Glu Ile Lys

100 105

<210> 35

<211> 119

<212> Protein

<213> Artificial sequence

<400> 35

Glu Val Gln Leu Leu Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Ala Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Leu Met Ala Thr Gly Gly Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 36

<211> 110

<212> Protein

<213> Artificial sequence

<400> 36

Gln Ser Val Leu Thr Gln Pro Ala Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asn Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Asn Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 37

<211> 113

<212> Protein

<213> Artificial sequence

<400> 37

Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Asp Cys Lys Ala Ser Gly Ile Thr Phe Ser Asn Ser

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Tyr Asp Gly Ser Lys Arg Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Phe

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Asn Asp Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

100 105 110

Ser

<210> 38

<211> 107

<212> Protein

<213> Artificial sequence

<400> 38

Glu Ile Val Leu Thr Gln Ser Pro Ala 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 Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Ser Asn Trp Pro Arg

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 39

<211> 120

<212> Protein

<213> Artificial sequence

<400> 39

Gln Val Gln Leu Val Gln Ser Gly Val Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Asn Pro Ser Asn Gly Gly Thr Asn Phe Asn Glu Lys Phe

50 55 60

Lys Asn Arg Val Thr Leu Thr Thr Asp Ser Ser Thr Thr Thr Ala Tyr

65 70 75 80

Met Glu Leu Lys Ser Leu Gln Phe Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Asp Tyr Arg Phe Asp Met Gly Phe Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 40

<211> 111

<212> Protein

<213> Artificial sequence

<400> 40

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Lys Gly Val Ser Thr Ser

20 25 30

Gly Tyr Ser Tyr Leu His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Arg Leu Leu Ile Tyr Leu Ala Ser Tyr Leu Glu Ser Gly Val Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln His Ser Arg

85 90 95

Asp Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 41

<211> 118

<212> Protein

<213> Artificial sequence

<400> 41

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 Ser

115

<210> 42

<211> 107

<212> Protein

<213> Artificial sequence

<400> 42

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

100 105

<210> 43

<211> 120

<212> Protein

<213> Artificial sequence

<400> 43

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 Ser Tyr

20 25 30

Ile Met Met Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Tyr Pro Ser Gly Gly Ile Thr Phe Tyr Ala Asp Thr 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 Ile Lys Leu Gly Thr Val Thr Thr Val Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 44

<211> 110

<212> Protein

<213> Artificial sequence

<400> 44

Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu

65 70 75 80

Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser

85 90 95

Ser Thr Arg Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu

100 105 110

<210> 45

<211> 123

<212> Protein

<213> Artificial sequence

<400> 45

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Thr Ser Gly Asp Thr Phe Ser Thr Tyr

20 25 30

Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys

85 90 95

Ala Arg Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val

100 105 110

Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 46

<211> 106

<212> Protein

<213> Artificial sequence

<400> 46

Glu Ile Val Leu Thr Gln Ser Pro Ala 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 Tyr

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile

35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro

65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Thr

85 90 95

Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 47

<211> 117

<212> Protein

<213> Artificial sequence

<400> 47

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Arg Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Gly Ile Asn Trp Asn Gly Gly Ser Thr Gly Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser 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 Gly Arg Ser Leu Leu Phe Asp Tyr Trp Gly Gln Gly Thr Leu

100 105 110

Val Thr Val Ser Arg

115

<210> 48

<211> 107

<212> Protein

<213> Artificial sequence

<400> 48

Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln Thr

1 5 10 15

Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser

20 25 30

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr Gly

35 40 45

Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser

50 55 60

Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp

65 70 75 80

Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His Val

85 90 95

Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105

<210> 49

<211> 119

<212> Protein

<213> Artificial sequence

<400> 49

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 Asp Phe Ser Arg Tyr

20 25 30

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Ile

35 40 45

Gly Glu Ile Asn Pro Asp Ser Ser Thr Ile Asn Tyr Ala Pro Ser Leu

50 55 60

Lys Asp Lys Phe Ile Ile Ser Arg Asp Asn Ala Lys Asn Ser 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 Pro Asp Gly Asn Tyr Trp Tyr Phe Asp Val Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 50

<211> 107

<212> Protein

<213> Artificial sequence

<400> 50

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 Lys Ala Ser Gln Asp Val Gly Ile Ala

20 25 30

Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile

35 40 45

Tyr Trp Ala Ser Thr Arg His Thr Gly Val Pro Asp 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 Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Tyr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 51

<211> 121

<212> Protein

<213> Artificial sequence

<400> 51

Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Val Phe

20 25 30

Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Thr Lys Thr Gly Glu Ala Thr Tyr Val Glu Glu Phe

50 55 60

Lys Gly Arg Phe Val Phe Ser Leu Asp Thr Ser Val Ser Thr Ala Tyr

65 70 75 80

Leu Gln Ile Ser Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Trp Asp Phe Tyr Asp Tyr Val Glu Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Thr Val Thr Val Ser Ser

115 120

<210> 52

<211> 108

<212> Protein

<213> Artificial sequence

<400> 52

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 Lys Ala Ser Gln Asn Val Gly Thr Asn

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 Tyr Arg Tyr Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Ile Ala Thr Tyr Tyr Cys His Gln Tyr Tyr Thr Tyr Pro Leu

85 90 95

Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 53

<211> 123

<212> Protein

<213> Artificial sequence

<400> 53

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 Tyr Thr Phe Thr Asn Tyr

20 25 30

Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Ala Asp Phe

50 55 60

Lys Arg Arg Phe Thr Phe Ser Leu Asp Thr Ser Lys Ser 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 Lys Tyr Pro His Tyr Tyr Gly Ser Ser His Trp Tyr Phe Asp Val

100 105 110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 54

<211> 107

<212> Protein

<213> Artificial sequence

<400> 54

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 Ser Ala Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile

35 40 45

Tyr Phe Thr Ser Ser Leu His 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 Ser Thr Val Pro Trp

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 55

<211> 121

<212> Protein

<213> Artificial sequence

<400> 55

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 Ser Ile Asn Gly Ser

20 25 30

Trp Ile Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Gly Ala Ile Trp Pro Phe Gly Gly Tyr Thr His 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 Trp Gly His Ser Thr Ser Pro Trp Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 56

<211> 107

<212> Protein

<213> Artificial sequence

<400> 56

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 Val Ile Arg Arg Ser

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Asn Leu Ala 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 Asn Thr Ser Pro Leu

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 57

<211> 120

<212> Protein

<213> Artificial sequence

<400> 57

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 Ile Ser Asp Tyr

20 25 30

Trp Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Gly Ile Thr Pro Ala Gly Gly Tyr 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 Phe Val Phe Phe Leu Pro Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 58

<211> 107

<212> Protein

<213> Artificial sequence

<400> 58

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 Gly Tyr Gly Asn Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 59

<211> 120

<212> Protein

<213> Artificial sequence

<400> 59

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ser Thr Leu Gly Leu Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 60

<211> 108

<212> Protein

<213> Artificial sequence

<400> 60

Glu Thr 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 Leu Gly 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 Pro 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 Tyr Ala Asp Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 61

<211> 120

<212> Protein

<213> Artificial sequence

<400> 61

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Ser Asn

20 25 30

Val Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Gly Val Ile Pro Ile Val Asp Ile Ala Asn Tyr Ala Gln Arg Phe

50 55 60

Lys Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Thr Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Leu Pro Arg Ala Phe Val Leu Asp Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 62

<211> 108

<212> Protein

<213> Artificial sequence

<400> 62

Glu Thr 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 Leu Gly 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 Pro 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 Tyr Ala Asp Ser Pro

85 90 95

Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys

100 105

<210> 63

<211> 117

<212> Protein

<213> Artificial sequence

<400> 63

Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Leu Gln Pro Ser Gln

1 5 10 15

Ser Leu Ser Ile Ser Cys Thr Val Ser Gly Phe Ser Leu Ala Thr Tyr

20 25 30

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met

50 55 60

Ser Arg Leu Ser Ile Thr Lys Asp Asn Ser Lys Ser Gln Val Phe Phe

65 70 75 80

Lys Met Asn Ser Leu Gln Ala Asp Asp Thr Ala Ile Tyr Phe Cys Ala

85 90 95

Lys Gln Ala Tyr Gly His Tyr Met Asp Tyr Trp Gly Gln Gly Thr Ser

100 105 110

Val Thr Val Ser Ser

115

<210> 64

<211> 112

<212> Protein

<213> Artificial sequence

<400> 64

Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Thr Arg Leu Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 65

<211> 117

<212> Protein

<213> Artificial sequence

<400> 65

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 Ser Phe Ala Thr Tyr

20 25 30

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Arg Gly Gly Ser Thr Asp Tyr Ser Ala Ala Phe Met

50 55 60

Ser Arg Leu Thr Ile Ser Lys Asp Asn Ser Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Phe Cys Ala

85 90 95

Lys Gln Ala Tyr Gly His Tyr Met Asp Tyr Trp Gly Gln Gly Thr Ser

100 105 110

Val Thr Val Ser Ser

115

<210> 66

<211> 112

<212> Protein

<213> Artificial sequence

<400> 66

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Asn Ile Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Arg Pro Gly Gln Ser

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly

85 90 95

Ser His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105 110

<210> 67

<211> 118

<212> Protein

<213> Artificial sequence

<400> 67

Glu Val Lys Leu Asp Glu Thr Gly Gly Gly Leu Val Gln Pro Gly Arg

1 5 10 15

Pro Met Lys Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Trp Met Asn Trp Val Arg Gln Ser Pro Glu Lys Gly Leu Glu Trp Val

35 40 45

Ala Gln Ile Arg Asn Lys Pro Tyr Asn Tyr Glu Thr Tyr Tyr Ser Asp

50 55 60

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Ser Ser

65 70 75 80

Val Tyr Leu Gln Met Asn Asn Leu Arg Val Glu Asp Met Gly Ile Tyr

85 90 95

Tyr Cys Thr Gly Ser Tyr Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Ser Val Thr Val Ser Ser

115

<210> 68

<211> 112

<212> Protein

<213> Artificial sequence

<400> 68

Asp Val Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly

1 5 10 15

Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu Arg Trp Tyr Leu Gln Lys Pro Gly Gln Ser

35 40 45

Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Phe Cys Ser Gln Ser

85 90 95

Thr His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 69

<211> 2

<212> Protein

<213> Artificial sequence

<400> 69

Ser Ser

1

<210> 70

<211> 2

<212> Protein

<213> Artificial sequence

<400> 70

Ala Ser

1

<210> 71

<211> 5

<212> Protein

<213> Artificial sequence

<400> 71

Gly Gly Gly Gly Ser

15

<210> 72

<211> 7

<212> Protein

<213> Artificial sequence

<400> 72

Gly Gly Gly Ser Ala Ala Ala

15

<210> 73

<211> 7

<212> Protein

<213> Artificial sequence

<400> 73

Gly Gly Gly Gly Ser Ala Ser

15

<210> 74

<211> 8

<212> Protein

<213> Artificial sequence

<400> 74

Gly Tyr Pro Gly Gly Gly Gly Ser

15

<210> 75

<211> 10

<212> Protein

<213> Artificial sequence

<400> 75

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10

<210> 76

<211> 12

<212> Protein

<213> Artificial sequence

<400> 76

Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro Ala Pro

1 5 10

<210> 77

<211> 12

<212> Protein

<213> Artificial sequence

<400> 77

Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala

1 5 10

<210> 78

<211> 12

<212> Protein

<213> Artificial sequence

<400> 78

Gly Lys Ser Ser Gly Ser Gly Ser Glu Ser Lys Ser

1 5 10

<210> 79

<211> 14

<212> Protein

<213> Artificial sequence

<400> 79

Gly Ser Thr Ser Gly Ser Gly Lys Ser Ser Glu Gly Lys Gly

1 5 10

<210> 80

<211> 14

<212> Protein

<213> Artificial sequence

<400> 80

Glu Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser

1 5 10

<210> 81

<211> 14

<212> Protein

<213> Artificial sequence

<400> 81

Gly Gly Gly Gly Ser Asp Lys Thr His Thr Ser Pro Pro Ser

1 5 10

<210> 82

<211> 15

<212> Protein

<213> Artificial sequence

<400> 82

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<210> 83

<211> 14

<212> Protein

<213> Artificial sequence

<400> 83

Gly Ala Ala Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10

<210> 84

<211> 15

<212> Protein

<213> Artificial sequence

<400> 84

Glu Ala Ala Ala Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

<210> 85

<211> 15

<212> Protein

<213> Artificial sequence

<400> 85

Gly Gly Gly Gly Ser Glu Ala Ala Ala Lys Gly Gly Gly Gly Ser

1 5 10 15

<210> 86

<211> 15

<212> Protein

<213> Artificial sequence

<400> 86

Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Gly Gly Gly Gly Ser

1 5 10 15

<210> 87

<211> 17

<212> Protein

<213> Artificial sequence

<400> 87

Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

1 5 10 15

Gly

<210> 88

<211> 18

<212> Protein

<213> Artificial sequence

<400> 88

Gly Ser Thr Ser Gly Ser Gly Lys Ser Ser Glu Gly Ser Gly Ser Thr

1 5 10 15

Lys Gly

<210> 89

<211> 19

<212> Protein

<213> Artificial sequence

<400> 89

Gly Ala Ala Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

1 5 10 15

Gly Gly Ser

<210> 90

<211> 20

<212> Protein

<213> Artificial sequence

<400> 90

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

1 5 10 15

Gly Gly Gly Ser

20

<210> 91

<211> 7

<212> Protein

<213> Artificial sequence

<400> 91

Asp Lys Thr His Thr Cys Pro

15

<210> 92

<211> 9

<212> Protein

<213> Artificial sequence

<400> 92

Glu Arg Lys Cys Cys Val Glu Cys Pro

15

<210> 93

<211> 60

<212> Protein

<213> Artificial sequence

<400> 93

Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro Arg Cys

1 5 10 15

Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro

20 25 30

Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro Glu

35 40 45

Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg

50 55 60

<210> 94

<211> 12

<212> Protein

<213> Artificial sequence

<400> 94

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro

1 5 10

<210> 95

<211> 5

<212> Protein

<213> Artificial sequence

<400> 95

Asp Lys Thr His Thr

15

<210> 96

<211> 12

<212> Protein

<213> Artificial sequence

<400> 96

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro

1 5 10

<210> 97

<211> 12

<212> Protein

<213> Artificial sequence

<400> 97

Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro

1 5 10

<210> 98

<211> 12

<212> Protein

<213> Artificial sequence

<400> 98

Gly Gly Gly Gly Ser Asp Lys Thr His Thr Cys Pro

1 5 10

<210> 99

<211> 12

<212> Protein

<213> Artificial sequence

<400> 99

Arg Gly Arg Gly Ser Asp Lys Thr His Thr Cys Pro

1 5 10

<210> 100

<211> 12

<212> Protein

<213> Artificial sequence

<400> 100

Asp Gly Asp Gly Ser Asp Lys Thr His Thr Cys Pro

1 5 10

<210> 101

<211> 13

<212> Protein

<213> Artificial sequence

<400> 101

Gly Arg Gly Arg Gly Ser Asp Lys Thr His Thr Cys Pro

1 5 10

<210> 102

<211> 15

<212> Protein

<213> Artificial sequence

<400> 102

Ala Ser Thr Arg Gly Arg Gly Ser Asp Lys Thr His Thr Cys Pro

1 5 10 15

<210> 103

<211> 15

<212> Protein

<213> Artificial sequence

<400> 103

Gly Gln Pro Asp Gly Asp Ala Ser Asp Lys Thr His Thr Cys Pro

1 5 10 15

<210> 104

<211> 107

<212> Protein

<213> Artificial sequence

<400> 104

Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu

1 5 10 15

Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe

20 25 30

Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln

35 40 45

Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser

50 55 60

Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu

65 70 75 80

Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser

85 90 95

Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 105

<211> 106

<212> Protein

<213> Artificial sequence

<400> 105

Gly Gln Pro Lys Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro

35 40 45

Val Lys Ala Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

100 105

<210> 106

<211> 106

<212> Protein

<213> Artificial sequence

<400> 106

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro

35 40 45

Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

100 105

<210> 107

<211> 106

<212> Protein

<213> Artificial sequence

<400> 107

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro

35 40 45

Ala Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Lys Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

100 105

<210> 108

<211> 106

<212> Protein

<213> Artificial sequence

<400> 108

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Lys Val Ala Trp Lys Ala Asp Gly Ser Pro

35 40 45

Val Asn Thr Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Ala Glu Cys Ser

100 105

<210> 109

<211> 106

<212> Protein

<213> Artificial sequence

<400> 109

Gly Gln Pro Lys Ala Ala Pro Thr Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

20 25 30

Phe Tyr Pro Gly Ala Val Lys Val Ala Trp Lys Ala Asp Ser Ser Pro

35 40 45

Ala Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Lys Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

100 105

<210> 110

<211> 105

<212> Protein

<213> Artificial sequence

<400> 110

Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu

1 5 10 15

Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro

20 25 30

Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly

35 40 45

Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr

50 55 60

Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His

65 70 75 80

Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val

85 90 95

Thr Lys Ser Phe Asn Arg Gly Glu Cys

100 105

<210> 111

<211> 103

<212> Protein

<213> Artificial sequence

<400> 111

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Lys Val Glu Pro Lys Ser Cys

100

<210> 112

<211> 98

<212> Protein

<213> Artificial sequence

<400> 112

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Thr Val

<210> 113

<211> 98

<212> Protein

<213> Artificial sequence

<400> 113

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val

<210> 114

<211> 98

<212> Protein

<213> Artificial sequence

<400> 114

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Arg Val

<210> 115

<211> 113

<212> Protein

<213> Artificial sequence

<400> 115

Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe

1 5 10 15

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

20 25 30

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe

35 40 45

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

50 55 60

Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr

65 70 75 80

Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

85 90 95

Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala

100 105 110

Lys

<210> 116

<211> 112

<212> Protein

<213> Artificial sequence

<400> 116

Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro

1 5 10 15

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

20 25 30

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn

35 40 45

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

50 55 60

Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val

65 70 75 80

Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

85 90 95

Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys

100 105 110

<210> 117

<211> 112

<212> Protein

<213> Artificial sequence

<400> 117

Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro

1 5 10 15

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

20 25 30

Cys Val Val Val Asp Val Ser His Glu Ala Pro Glu Val Gln Phe Asn

35 40 45

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

50 55 60

Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val

65 70 75 80

Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

85 90 95

Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys

100 105 110

<210> 118

<211> 112

<212> Protein

<213> Artificial sequence

<400> 118

Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro

1 5 10 15

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

20 25 30

Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Lys

35 40 45

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

50 55 60

Glu Glu Gln Tyr Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val

65 70 75 80

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

85 90 95

Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys

100 105 110

<210> 119

<211> 110

<212> Protein

<213> Artificial sequence

<400> 119

Ala Pro Glu Phe Leu 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 Gln Glu Asp Pro Glu Val Gln 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 Phe 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

Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys

100 105 110

<210> 120

<211> 107

<212> Protein

<213> Artificial sequence

<400> 120

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 Lys

100 105

<210> 121

<211> 107

<212> Protein

<213> Artificial sequence

<400> 121

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu

1 5 10 15

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

20 25 30

Tyr Pro Ser Asp Ile Ser Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

35 40 45

Asn Asn Tyr Lys Thr Thr Pro Pro Met 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 Lys

100 105

<210> 122

<211> 107

<212> Protein

<213> Artificial sequence

<400> 122

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu

1 5 10 15

Glu Met 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 Ser Gly Gln Pro Glu

35 40 45

Asn Asn Tyr Asn Thr Thr Pro Pro Met 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 Ile Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn Arg Phe

85 90 95

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

100 105

<210> 123

<211> 107

<212> Protein

<213> Artificial sequence

<400> 123

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu

1 5 10 15

Glu Met 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 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu 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 Leu Gly Lys

100 105

<210> 124

<211> 106

<212> Protein

<213> Artificial sequence

<400> 124

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> 125

<211> 106

<212> Protein

<213> Artificial sequence

<400> 125

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu

1 5 10 15

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

20 25 30

Tyr Pro Ser Asp Ile Ser Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

35 40 45

Asn Asn Tyr Lys Thr Thr Pro Pro Met 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> 126

<211> 106

<212> Protein

<213> Artificial sequence

<400> 126

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu

1 5 10 15

Glu Met 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 Ser Gly Gln Pro Glu

35 40 45

Asn Asn Tyr Asn Thr Thr Pro Pro Met 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 Ile Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn Arg Phe

85 90 95

Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

100 105

<210> 127

<211> 106

<212> Protein

<213> Artificial sequence

<400> 127

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu

1 5 10 15

Glu Met 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 Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu 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 Leu Gly

100 105

<210> 128

<211> 107

<212> Protein

<213> Artificial sequence

<400> 128

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 Ala

100 105

<210> 129

<211> 258

<212> Protein

<213> Artificial sequence

<400> 129

Val Pro Arg Trp Arg Gln Gln Trp Ser Gly Pro Gly Thr Thr Lys Arg

1 5 10 15

Phe Pro Glu Thr Val Leu Ala Arg Cys Val Lys Tyr Thr Glu Ile His

20 25 30

Pro Glu Met Arg His Val Asp Cys Gln Ser Val Trp Asp Ala Phe Lys

35 40 45

Gly Ala Phe Ile Ser Lys His Pro Cys Asn Ile Thr Glu Glu Asp Tyr

50 55 60

Gln Pro Leu Met Lys Leu Gly Thr Gln Thr Val Pro Cys Asn Lys Ile

65 70 75 80

Leu Leu Trp Ser Arg Ile Lys Asp Leu Ala His Gln Phe Thr Gln Val

85 90 95

Gln Arg Asp Met Phe Thr Leu Glu Asp Thr Leu Leu Gly Tyr Leu Ala

100 105 110

Asp Asp Leu Thr Trp Cys Gly Glu Phe Asn Thr Ser Lys Ile Asn Tyr

115 120 125

Gln Ser Cys Pro Asp Trp Arg Lys Asp Cys Ser Asn Asn Pro Val Ser

130 135 140

Val Phe Trp Lys Thr Val Ser Arg Arg Phe Ala Glu Ala Ala Cys Asp

145 150 155 160

Val Val His Val Met Leu Asn Gly Ser Arg Ser Lys Ile Phe Asp Lys

165 170 175

Asn Ser Thr Phe Gly Ser Val Glu Val His Asn Leu Gln Pro Glu Lys

180 185 190

Val Gln Thr Leu Glu Ala Trp Val Ile His Gly Gly Arg Glu Asp Ser

195 200 205

Arg Asp Leu Cys Gln Asp Pro Thr Ile Lys Glu Leu Glu Ser Ile Ile

210 215 220

Ser Lys Arg Asn Ile Gln Phe Ser Cys Lys Asn Ile Tyr Arg Pro Asp

225 230 235 240

Lys Phe Leu Gln Cys Val Lys Asn Pro Glu Asp Ser Ser Cys Thr Ser

245 250 255

Glu Ile

<210> 130

<211> 54

<212> Protein

<213> Artificial sequence

<400> 130

Met Leu Gln Met Ala Gly Gln Cys Ser Gln Asn Glu Tyr Phe Asp Ser

1 5 10 15

Leu Leu His Ala Cys Ile Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr

20 25 30

Pro Pro Leu Thr Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr Asn Ser

35 40 45

Val Lys Gly Thr Asn Ala

50

<210> 131

<211> 124

<212> Protein

<213> Artificial sequence

<400> 131

Met His Val Ala Gln Pro Ala Val Val Leu Ala Ser Ser Arg Gly Ile

1 5 10 15

Ala Ser Phe Val Cys Glu Tyr Ala Ser Pro Gly Lys Ala Thr Glu Val

20 25 30

Arg Val Thr Val Leu Arg Gln Ala Asp Ser Gln Val Thr Glu Val Cys

35 40 45

Ala Ala Thr Tyr Met Met Gly Asn Glu Leu Thr Phe Leu Asp Asp Ser

50 55 60

Ile Cys Thr Gly Thr Ser Ser Gly Asn Gln Val Asn Leu Thr Ile Gln

65 70 75 80

Gly Leu Arg Ala Met Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu

85 90 95

Met Tyr Pro Pro Pro Tyr Tyr Leu Gly Ile Gly Asn Gly Thr Gln Ile

100 105 110

Tyr Val Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp

115 120

<210> 132

<211> 422

<212> Protein

<213> Artificial sequence

<400> 132

Val Pro Val Val Trp Ala Gln Glu Gly Ala Pro Ala Gln Leu Pro Cys

1 5 10 15

Ser Pro Thr Ile Pro Leu Gln Asp Leu Ser Leu Leu Arg Arg Ala Gly

20 25 30

Val Thr Trp Gln His Gln Pro Asp Ser Gly Pro Pro Ala Ala Ala Pro

35 40 45

Gly His Pro Leu Ala Pro Gly Pro His Pro Ala Ala Pro Ser Ser Trp

50 55 60

Gly Pro Arg Pro Arg Arg Tyr Thr Val Leu Ser Val Gly Pro Gly Gly

65 70 75 80

Leu Arg Ser Gly Arg Leu Pro Leu Gln Pro Arg Val Gln Leu Asp Glu

85 90 95

Arg Gly Arg Gln Arg Gly Asp Phe Ser Leu Trp Leu Arg Pro Ala Arg

100 105 110

Arg Ala Asp Ala Gly Glu Tyr Arg Ala Ala Val His Leu Arg Asp Arg

115 120 125

Ala Leu Ser Cys Arg Leu Arg Leu Arg Leu Gly Gln Ala Ser Met Thr

130 135 140

Ala Ser Pro Pro Gly Ser Leu Arg Ala Ser Asp Trp Val Ile Leu Asn

145 150 155 160

Cys Ser Phe Ser Arg Pro Asp Arg Pro Ala Ser Val His Trp Phe Arg

165 170 175

Asn Arg Gly Gln Gly Arg Val Pro Val Arg Glu Ser Pro His His His

180 185 190

Leu Ala Glu Ser Phe Leu Phe Leu Pro Gln Val Ser Pro Met Asp Ser

195 200 205

Gly Pro Trp Gly Cys Ile Leu Thr Tyr Arg Asp Gly Phe Asn Val Ser

210 215 220

Ile Met Tyr Asn Leu Thr Val Leu Gly Leu Glu Pro Pro Thr Pro Leu

225 230 235 240

Thr Val Tyr Ala Gly Ala Gly Ser Arg Val Gly Leu Pro Cys Arg Leu

245 250 255

Pro Ala Gly Val Gly Thr Arg Ser Phe Leu Thr Ala Lys Trp Thr Pro

260 265 270

Pro Gly Gly Gly Pro Asp Leu Leu Val Thr Gly Asp Asn Gly Asp Phe

275 280 285

Thr Leu Arg Leu Glu Asp Val Ser Gln Ala Gln Ala Gly Thr Tyr Thr

290 295 300

Cys His Ile His Leu Gln Glu Gln Gln Leu Asn Ala Thr Val Thr Leu

305 310 315 320

Ala Ile Ile Thr Val Thr Pro Lys Ser Phe Gly Ser Pro Gly Ser Leu

325 330 335

Gly Lys Leu Leu Cys Glu Val Thr Pro Val Ser Gly Gln Glu Arg Phe

340 345 350

Val Trp Ser Ser Leu Asp Thr Pro Ser Gln Arg Ser Phe Ser Gly Pro

355 360 365

Trp Leu Glu Ala Gln Glu Ala Gln Leu Leu Ser Gln Pro Trp Gln Cys

370 375 380

Gln Leu Tyr Gln Gly Glu Arg Leu Leu Gly Ala Ala Val Tyr Phe Thr

385 390 395 400

Glu Leu Ser Ser Pro Gly Ala Gln Arg Ser Gly Arg Ala Pro Gly Ala

405 410 415

Leu Pro Ala Gly His Leu

420

<210> 133

<211> 120

<212> Protein

<213> Artificial sequence

<400> 133

Met Met Thr Gly Thr Ile Glu Thr Thr Gly Asn Ile Ser Ala Glu Lys

1 5 10 15

Gly Gly Ser Ile Ile Leu Gln Cys His Leu Ser Ser Thr Thr Ala Gln

20 25 30

Val Thr Gln Val Asn Trp Glu Gln Gln Asp Gln Leu Leu Ala Ile Cys

35 40 45

Asn Ala Asp Leu Gly Trp His Ile Ser Pro Ser Phe Lys Asp Arg Val

50 55 60

Ala Pro Gly Pro Gly Leu Gly Leu Thr Leu Gln Ser Leu Thr Val Asn

65 70 75 80

Asp Thr Gly Glu Tyr Phe Cys Ile Tyr His Thr Tyr Pro Asp Gly Thr

85 90 95

Tyr Thr Gly Arg Ile Phe Leu Glu Val Leu Glu Ser Ser Val Ala Glu

100 105 110

His Gly Ala Arg Phe Gln Ile Pro

115 120

<210> 134

<211> 150

<212> Protein

<213> Artificial sequence

<400> 134

Pro Gly Trp Phe Leu Asp Ser Pro Asp Arg Pro Trp Asn Pro Pro Thr

1 5 10 15

Phe Ser Pro Ala Leu Leu Val Val Thr Glu Gly Asp Asn Ala Thr Phe

20 25 30

Thr Cys Ser Phe Ser Asn Thr Ser Glu Ser Phe Val Leu Asn Trp Tyr

35 40 45

Arg Met Ser Pro Ser Asn Gln Thr Asp Lys Leu Ala Ala Phe Pro Glu

50 55 60

Asp Arg Ser Gln Pro Gly Gln Asp Cys Arg Phe Arg Val Thr Gln Leu

65 70 75 80

Pro Asn Gly Arg Asp Phe His Met Ser Val Val Arg Ala Arg Arg Asn

85 90 95

Asp Ser Gly Thr Tyr Leu Cys Gly Ala Ile Ser Leu Ala Pro Lys Ala

100 105 110

Gln Ile Lys Glu Ser Leu Arg Ala Glu Leu Arg Val Thr Glu Arg Arg

115 120 125

Ala Glu Val Pro Thr Ala His Pro Ser Pro Ser Pro Arg Pro Ala Gly

130 135 140

Gln Phe Gln Thr Leu Val

145 150

<210> 135

<211> 220

<212> Protein

<213> Artificial sequence

<400> 135

Phe Thr Val Thr Val Pro Lys Asp Leu Tyr Val Val Glu Tyr Gly Ser

1 5 10 15

Asn Met Thr Ile Glu Cys Lys Phe Pro Val Glu Lys Gln Leu Asp Leu

20 25 30

Ala Ala Leu Ile Val Tyr Trp Glu Met Glu Asp Lys Asn Ile Ile Gln

35 40 45

Phe Val His Gly Glu Glu Asp Leu Lys Val Gln His Ser Ser Tyr Arg

50 55 60

Gln Arg Ala Arg Leu Leu Lys Asp Gln Leu Ser Leu Gly Asn Ala Ala

65 70 75 80

Leu Gln Ile Thr Asp Val Lys Leu Gln Asp Ala Gly Val Tyr Arg Cys

85 90 95

Met Ile Ser Tyr Gly Gly Ala Asp Tyr Lys Arg Ile Thr Val Lys Val

100 105 110

Asn Ala Pro Tyr Asn Lys Ile Asn Gln Arg Ile Leu Val Val Asp Pro

115 120 125

Val Thr Ser Glu His Glu Leu Thr Cys Gln Ala Glu Gly Tyr Pro Lys

130 135 140

Ala Glu Val Ile Trp Thr Ser Ser Asp His Gln Val Leu Ser Gly Lys

145 150 155 160

Thr Thr Thr Thr Asn Ser Lys Arg Glu Glu Lys Leu Phe Asn Val Thr

165 170 175

Ser Thr Leu Arg Ile Asn Thr Thr Thr Asn Glu Ile Phe Tyr Cys Thr

180 185 190

Phe Arg Arg Leu Asp Pro Glu Glu Asn His Thr Ala Glu Leu Val Ile

195 200 205

Pro Glu Leu Pro Leu Ala His Pro Pro Asn Glu Arg

210 215 220

<210> 136

<211> 204

<212> Protein

<213> Artificial sequence

<400> 136

Ser Gly Pro Val Lys Glu Leu Val Gly Ser Val Gly Gly Ala Val Thr

1 5 10 15

Phe Pro Leu Lys Ser Lys Val Lys Gln Val Asp Ser Ile Val Trp Thr

20 25 30

Phe Asn Thr Thr Pro Leu Val Thr Ile Gln Pro Glu Gly Gly Thr Ile

35 40 45

Ile Val Thr Gln Asn Arg Asn Arg Glu Arg Val Asp Phe Pro Asp Gly

50 55 60

Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys Lys Asn Asp Ser Gly Ile

65 70 75 80

Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu Gln Gln Pro Ser Thr Gln

85 90 95

Glu Tyr Val Leu His Val Tyr Glu His Leu Ser Lys Pro Lys Val Thr

100 105 110

Met Gly Leu Gln Ser Asn Lys Asn Gly Thr Cys Val Thr Asn Leu Thr

115 120 125

Cys Cys Met Glu His Gly Glu Glu Asp Val Ile Tyr Thr Trp Lys Ala

130 135 140

Leu Gly Gln Ala Ala Asn Glu Ser His Asn Gly Ser Ile Leu Pro Ile

145 150 155 160

Ser Trp Arg Trp Gly Glu Ser Asp Met Thr Phe Ile Cys Val Ala Arg

165 170 175

Asn Pro Val Ser Arg Asn Phe Ser Ser Pro Ile Leu Ala Arg Lys Leu

180 185 190

Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser Ser Met

195 200

<210> 137

<211> 651

<212> Protein

<213> Artificial sequence

<400> 137

Lys Leu Thr Ile Glu Ser Thr Pro Phe Asn Val Ala Glu Gly Lys Glu

1 5 10 15

Val Leu Leu Leu Val His Asn Leu Pro Gln His Leu Phe Gly Tyr Ser

20 25 30

Trp Tyr Lys Gly Glu Arg Val Asp Gly Asn Arg Gln Ile Ile Gly Tyr

35 40 45

Val Ile Gly Thr Gln Gln Ala Thr Pro Gly Pro Ala Tyr Ser Gly Arg

50 55 60

Glu Ile Ile Tyr Pro Asn Ala Ser Leu Leu Ile Gln Asn Ile Ile Gln

65 70 75 80

Asn Asp Thr Gly Phe Tyr Thr Leu His Val Ile Lys Ser Asp Leu Val

85 90 95

Asn Glu Glu Ala Thr Gly Gln Phe Arg Val Tyr Pro Glu Leu Pro Lys

100 105 110

Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro Val Glu Asp Lys Asp Ala

115 120 125

Val Ala Phe Thr Cys Glu Pro Glu Thr Gln Asp Ala Thr Tyr Leu Trp

130 135 140

Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg Leu Gln Leu Ser

145 150 155 160

Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn Val Thr Arg Asn Asp Thr

165 170 175

Ala Ser Tyr Lys Cys Glu Thr Gln Asn Pro Val Ser Ala Arg Arg Ser

180 185 190

Asp Ser Val Ile Leu Asn Val Leu Tyr Gly Pro Asp Ala Pro Thr Ile

195 200 205

Ser Pro Leu Asn Thr Ser Tyr Arg Ser Gly Glu Asn Leu Asn Leu Ser

210 215 220

Cys His Ala Ala Ser Asn Pro Pro Ala Gln Tyr Ser Trp Phe Val Asn

225 230 235 240

Gly Thr Phe Gln Gln Ser Thr Gln Glu Leu Phe Ile Pro Asn Ile Thr

245 250 255

Val Asn Asn Ser Gly Ser Tyr Thr Cys Gln Ala His Asn Ser Asp Thr

260 265 270

Gly Leu Asn Arg Thr Thr Val Thr Thr Ile Thr Val Tyr Ala Glu Pro

275 280 285

Pro Lys Pro Phe Ile Thr Ser Asn Asn Ser Asn Pro Val Glu Asp Glu

290 295 300

Asp Ala Val Ala Leu Thr Cys Glu Pro Glu Ile Gln Asn Thr Thr Tyr

305 310 315 320

Leu Trp Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg Leu Gln

325 330 335

Leu Ser Asn Asp Asn Arg Thr Leu Thr Leu Leu Ser Val Thr Arg Asn

340 345 350

Asp Val Gly Pro Tyr Glu Cys Gly Ile Gln Asn Lys Leu Ser Val Asp

355 360 365

His Ser Asp Pro Val Ile Leu Asn Val Leu Tyr Gly Pro Asp Asp Pro

370 375 380

Thr Ile Ser Pro Ser Tyr Thr Tyr Tyr Arg Pro Gly Val Asn Leu Ser

385 390 395 400

Leu Ser Cys His Ala Ala Ser Asn Pro Pro Ala Gln Tyr Ser Trp Leu

405 410 415

Ile Asp Gly Asn Ile Gln Gln His Thr Gln Glu Leu Phe Ile Ser Asn

420 425 430

Ile Thr Glu Lys Asn Ser Gly Leu Tyr Thr Cys Gln Ala Asn Asn Ser

435 440 445

Ala Ser Gly His Ser Arg Thr Thr Val Lys Thr Ile Thr Val Ser Ala

450 455 460

Glu Leu Pro Lys Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro Val Glu

465 470 475 480

Asp Lys Asp Ala Val Ala Phe Thr Cys Glu Pro Glu Ala Gln Asn Thr

485 490 495

Thr Tyr Leu Trp Trp Val Asn Gly Gln Ser Leu Pro Val Ser Pro Arg

500 505 510

Leu Gln Leu Ser Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn Val Thr

515 520 525

Arg Asn Asp Ala Arg Ala Tyr Val Cys Gly Ile Gln Asn Ser Val Ser

530 535 540

Ala Asn Arg Ser Asp Pro Val Thr Leu Asp Val Leu Tyr Gly Pro Asp

545 550 555 560

Thr Pro Ile Ile Ser Pro Pro Asp Ser Ser Tyr Leu Ser Gly Ala Asn

565 570 575

Leu Asn Leu Ser Cys His Ser Ala Ser Asn Pro Ser Pro Gln Tyr Ser

580 585 590

Trp Arg Ile Asn Gly Ile Pro Gln Gln His Thr Gln Val Leu Phe Ile

595 600 605

Ala Lys Ile Thr Pro Asn Asn Asn Gly Thr Tyr Ala Cys Phe Val Ser

610 615 620

Asn Leu Ala Thr Gly Arg Asn Asn Ser Ile Val Lys Ser Ile Thr Val

625 630 635 640

Ser Ala Ser Gly Thr Ser Pro Gly Leu Ser Ala

645 650

<210> 138

<211> 104

<212> Protein

<213> Artificial sequence

<400> 138

Asp Gly Asn Glu Glu Met Gly Gly Ile Thr Gln Thr Pro Tyr Lys Val

1 5 10 15

Ser Ile Ser Gly Thr Thr Val Ile Leu Thr Cys Pro Gln Tyr Pro Gly

20 25 30

Ser Glu Ile Leu Trp Gln His Asn Asp Lys Asn Ile Gly Gly Asp Glu

35 40 45

Asp Asp Lys Asn Ile Gly Ser Asp Glu Asp His Leu Ser Leu Lys Glu

50 55 60

Phe Ser Glu Leu Glu Gln Ser Gly Tyr Tyr Val Cys Tyr Pro Arg Gly

65 70 75 80

Ser Lys Pro Glu Asp Ala Asn Phe Tyr Leu Tyr Leu Arg Ala Arg Val

85 90 95

Cys Glu Asn Cys Met Glu Met Asp

100

<210> 139

<211> 192

<212> Protein

<213> Artificial sequence

<400> 139

Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro

1 5 10 15

Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln

20 25 30

Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu

35 40 45

Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr

50 55 60

Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu

65 70 75 80

Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln

85 90 95

Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys

100 105 110

His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn

115 120 125

Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro

130 135 140

Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Phe

145 150 155 160

Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln

165 170 175

Gly Leu Ala Val Ser Thr Ile Ser Ser Phe Phe Pro Pro Gly Tyr Gln

180 185 190

<210> 140

<211> 112

<212> Protein

<213> Artificial sequence

<400> 140

Ala Leu Asp Thr Asn Tyr Cys Phe Ser Ser Thr Glu Lys Asn Cys Cys

1 5 10 15

Val Arg Gln Leu Tyr Ile Asp Phe Arg Lys Asp Leu Gly Trp Lys Trp

20 25 30

Ile His Glu Pro Lys Gly Tyr His Ala Asn Phe Cys Leu Gly Pro Cys

35 40 45

Pro Tyr Ile Trp Ser Leu Asp Thr Gln Tyr Ser Lys Val Leu Ala Leu

50 55 60

Tyr Asn Gln His Asn Pro Gly Ala Ser Ala Ala Pro Cys Cys Val Pro

65 70 75 80

Gln Ala Leu Glu Pro Leu Pro Ile Val Tyr Tyr Val Gly Arg Lys Pro

85 90 95

Lys Val Glu Gln Leu Ser Asn Met Ile Val Arg Ser Cys Lys Cys Ser

100 105 110

<210> 141

<211> 112

<212> Protein

<213> Artificial sequence

<400> 141

Ala Leu Asp Ala Ala Tyr Cys Phe Arg Asn Val Gln Asp Asn Cys Cys

1 5 10 15

Leu Arg Pro Leu Tyr Ile Asp Phe Lys Arg Asp Leu Gly Trp Lys Trp

20 25 30

Ile His Glu Pro Lys Gly Tyr Asn Ala Asn Phe Cys Ala Gly Ala Cys

35 40 45

Pro Tyr Leu Trp Ser Ser Asp Thr Gln His Ser Arg Val Leu Ser Leu

50 55 60

Tyr Asn Thr Ile Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val Ser

65 70 75 80

Gln Asp Leu Glu Pro Leu Thr Ile Leu Tyr Tyr Ile Gly Lys Thr Pro

85 90 95

Lys Ile Glu Gln Leu Ser Asn Met Ile Val Lys Ser Cys Lys Cys Ser

100 105 110

<210> 142

<211> 112

<212> Protein

<213> Artificial sequence

<400> 142

Ala Leu Asp Thr Asn Tyr Cys Phe Arg Asn Leu Glu Glu Asn Cys Cys

1 5 10 15

Val Arg Pro Leu Tyr Ile Asp Phe Arg Gln Asp Leu Gly Trp Lys Trp

20 25 30

Val His Glu Pro Lys Gly Tyr Tyr Ala Asn Phe Cys Ser Gly Pro Cys

35 40 45

Pro Tyr Leu Arg Ser Ala Asp Thr Thr His Ser Thr Val Leu Gly Leu

50 55 60

Tyr Asn Thr Leu Asn Pro Glu Ala Ser Ala Ser Pro Cys Cys Val Pro

65 70 75 80

Gln Asp Leu Glu Pro Leu Thr Ile Leu Tyr Tyr Val Gly Arg Thr Pro

85 90 95

Lys Val Glu Gln Leu Ser Asn Met Val Val Lys Ser Cys Lys Cys Ser

100 105 110

<210> 143

<211> 206

<212> Protein

<213> Artificial sequence

<400> 143

Ala Pro Met Ala Glu Gly Gly Gly Gln Asn His His Glu Val Val Lys

1 5 10 15

Phe Met Asp Val Tyr Gln Arg Ser Tyr Cys His Pro Ile Glu Thr Leu

20 25 30

Val Asp Ile Phe Gln Glu Tyr Pro Asp Glu Ile Glu Tyr Ile Phe Lys

35 40 45

Pro Ser Cys Val Pro Leu Met Arg Cys Gly Gly Cys Cys Asn Asp Glu

50 55 60

Gly Leu Glu Cys Val Pro Thr Glu Glu Ser Asn Ile Thr Met Gln Ile

65 70 75 80

Met Arg Ile Lys Pro His Gln Gly Gln His Ile Gly Glu Met Ser Phe

85 90 95

Leu Gln His Asn Lys Cys Glu Cys Arg Pro Lys Lys Asp Arg Ala Arg

100 105 110

Gln Glu Lys Lys Ser Val Arg Gly Lys Gly Lys Gly Gln Lys Arg Lys

115 120 125

Arg Lys Lys Ser Arg Tyr Lys Ser Trp Ser Val Tyr Val Gly Ala Arg

130 135 140

Cys Cys Leu Met Pro Trp Ser Leu Pro Gly Pro His Pro Cys Gly Pro

145 150 155 160

Cys Ser Glu Arg Arg Lys His Leu Phe Val Gln Asp Pro Gln Thr Cys

165 170 175

Lys Cys Ser Cys Lys Asn Thr Asp Ser Arg Cys Lys Ala Arg Gln Leu

180 185 190

Glu Leu Asn Glu Arg Thr Cys Arg Cys Asp Lys Pro Arg Arg

195 200 205

<210> 144

<211> 159

<212> Protein

<213> Artificial sequence

<400> 144

Pro Gly Gln Gly Thr Gln Ser Glu Asn Ser Cys Thr His Phe Pro Gly

1 5 10 15

Asn Leu Pro Asn Met Leu Arg Asp Leu Arg Asp Ala Phe Ser Arg Val

20 25 30

Lys Thr Phe Phe Gln Met Lys Asp Gln Leu Asp Asn Leu Leu Leu Lys

35 40 45

Glu Ser Leu Leu Glu Asp Phe Lys Gly Tyr Leu Gly Cys Gln Ala Leu

50 55 60

Ser Glu Met Ile Gln Phe Tyr Leu Glu Glu Val Met Pro Gln Ala Glu

65 70 75 80

Asn Gln Asp Pro Asp Ile Lys Ala His Val Asn Ser Leu Gly Glu Asn

85 90 95

Leu Lys Thr Leu Arg Leu Arg Leu Arg Arg Cys His Arg Phe Leu Pro

100 105 110

Cys Glu Asn Lys Ser Lys Ala Val Glu Gln Val Lys Asn Ala Phe Asn

115 120 125

Lys Leu Gln Glu Lys Gly Ile Tyr Lys Ala Met Ser Glu Phe Asp Ile

130 135 140

Phe Ile Asn Tyr Ile Glu Ala Tyr Met Thr Met Lys Ile Arg Asn

145 150 155

<210> 145

<211> 214

<212> Protein

<213> Artificial sequence

<400> 145

His Gly Thr Glu Leu Pro Ser Pro Pro Ser Val Trp Phe Glu Ala Glu

1 5 10 15

Phe Phe His His Ile Leu His Trp Thr Pro Ile Pro Asn Gln Ser Glu

20 25 30

Ser Thr Cys Tyr Glu Val Ala Leu Leu Arg Tyr Gly Ile Glu Ser Trp

35 40 45

Asn Ser Ile Ser Asn Cys Ser Gln Thr Leu Ser Tyr Asp Leu Thr Ala

50 55 60

Val Thr Leu Asp Leu Tyr His Ser Asn Gly Tyr Arg Ala Arg Val Arg

65 70 75 80

Ala Val Asp Gly Ser Arg His Ser Asn Trp Thr Val Thr Asn Thr Arg

85 90 95

Phe Ser Val Asp Glu Val Thr Leu Thr Val Gly Ser Val Asn Leu Glu

100 105 110

Ile His Asn Gly Phe Ile Leu Gly Lys Ile Gln Leu Pro Arg Pro Lys

115 120 125

Met Ala Pro Ala Asn Asp Thr Tyr Glu Ser Ile Phe Ser His Phe Arg

130 135 140

Glu Tyr Glu Ile Ala Ile Arg Lys Val Pro Gly Asn Phe Thr Phe Thr

145 150 155 160

His Lys Lys Val Lys His Glu Asn Phe Ser Leu Leu Thr Ser Gly Glu

165 170 175

Val Gly Glu Phe Cys Val Gln Val Lys Pro Ser Val Ala Ser Arg Ser

180 185 190

Asn Lys Gly Met Trp Ser Lys Glu Glu Cys Ile Ser Leu Thr Arg Gln

195 200 205

Tyr Phe Thr Val Thr Asn

210

<210> 146

<211> 120

<212> Protein

<213> Artificial sequence

<400> 146

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 Asn Ile Lys Asp Thr

20 25 30

Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Arg Ile Tyr Pro Thr Asn Gly Tyr Thr Arg 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

Ser Arg Trp Gly Gly Asp Gly Phe Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 147

<211> 107

<212> Protein

<213> Artificial sequence

<400> 147

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 Asn 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 Arg 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 His Tyr Thr Thr Pro Pro

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<210> 148

<211> 119

<212> Protein

<213> Artificial sequence

<400> 148

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 Thr Asp Tyr

20 25 30

Thr Met Asp Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Asp Val Asn Pro Asn Ser Gly Gly Ser Ile Tyr Asn Gln Arg Phe

50 55 60

Lys Gly Arg Phe Thr Leu Ser Val Asp Arg 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 Asn Leu Gly Pro Ser Phe Tyr Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser

115

<210> 149

<211> 107

<212> Protein

<213> Artificial sequence

<400> 149

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 Lys Ala Ser Gln Asp Val Ser Ile Gly

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 Tyr Arg Tyr 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 Tyr Tyr Ile Tyr Pro Tyr

85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100 105

<---

Claims (99)

1. A bispecific antibody that binds two different epitopes, containing two heavy chains and two light chains, characterized in that the two heavy chains and two light chains are two identical hybrid heavy chains and two identical hybrid light chains, where the two hybrid heavy chains form a pair and a hybrid light chain and a hybrid heavy chain form a pair; wherein the hybrid heavy chain includes the variable region of the heavy chain of antibody a (VHa), the first constant region CH1, the variable region 1 of antibody b and an Fc fragment, the variable region 1 of antibody b is linked to CH1 linker 2 or a peptide bond, and to Fc linker 3 or a peptide bond; The hybrid light chain of a bispecific antibody includes an antibody light chain variable region a (VLa), a light chain constant region CL, and an antibody b variable region 2, wherein antibody b variable region 2 is linked to the C-terminus of CL via linker 1 or a peptide bond, wherein (1) antibody b variable region 1 is an antibody b heavy chain variable region (VHb), and antibody b variable region 2 is an antibody b light chain variable region (VLb); or (2) antibody b variable region 1 is the antibody b light chain variable region (VLb), and antibody b variable region 2 is the antibody b heavy chain variable region (VHb), wherein the VHa-VLa pair targets the first epitope, and the VHb-VLb pair targets the second epitope, and wherein linker 1 and linker 2 are SEQ ID NO: 85. 2. Bispecific antibody according to claim 1, where the bispecific antibody is a tetravalent symmetric bispecific antibody with the structure F(ab) ₂ -(Fv) ₂ -Fc, where F(ab) ₂ includes two VHa, two CH1, two VLa and two CL; (Fv) ₂ includes two VHb and two VLb, where the positions of VHb and VLb are interchangeable; and the Fc portion includes a hinge region, a second CH2 constant region, and a third CH3 constant region. 3. The bispecific antibody according to claim 1, wherein the VHa-VLa pair forms one or more interchain disulfide bonds and the VHb-VLb pair forms one or more interchain disulfide bonds. 4. Bispecific antibody according to any one of paragraphs. 1-3, wherein antibody b variable region 1 is an antibody b light chain variable region (VLb) and antibody b variable region 2 is an antibody b heavy chain variable region (VHb). 5. Bispecific antibody according to any one of paragraphs. 1-4, where linkers 1-3 may be the same or different from each other. 6. Bispecific antibody according to any one of paragraphs. 1-5, wherein the CL sequence is selected from the group consisting of SEQ ID NO: 104-110. 7. Bispecific antibody according to any one of paragraphs. 1-6, wherein the CH1 sequence is selected from the group consisting of SEQ ID NO: 111-114. 8. Bispecific antibody according to any one of paragraphs. 1-7, wherein the CH2 sequence is selected from the group consisting of SEQ ID NO: 115-119. 9. Bispecific antibody according to any one of paragraphs. 1-8, wherein the CH3 sequence is selected from the group consisting of SEQ ID NO: 120-128. 10. Bispecific antibody according to any one of paragraphs. 1-9, wherein the first epitope and the second epitope are each independently selected from the group consisting of: immune cell surface antigens, tumor antigens, viruses, bacteria, endotoxins, cytokines, or combinations thereof. 11. Bispecific antibody according to any one of paragraphs. 1-10, where the first epitope and/or the second epitope is selected from PD-L1, PD-1, VEGFA, IL-10, IL-10R, BCMA, VEGF, TGF-β, CTLA-4, LAG-3, TIGIT, CEA, CD38, SLAMF7, B7-H3, Her2, EpCAM, CD19, CD20, CD30, CD33, CD47, CD52, CD133, EGFR, GD2, GD3, GM2, RANKL, CD3 and/or CD16a. 12. Bispecific antibody according to any one of paragraphs. 1-11, where the first epitope and/or the second epitope is selected from SEQ ID NO: 129-145. 13. Bispecific antibody according to any one of paragraphs. 1-12, where the first epitope and the second epitope are selected from the group consisting of: PD-L1 and VEGF, PD-1 and VEGF, PD-L1 and TGF-β, PD-1 and TGF-β, PD-1 and CTLA-4, PD-1 and LAG-3, PD-1 and TIGIT, PD-1 and IL-10, SLAMF7 and CD16a, and Her2 and Her2. 14. Bispecific antibody according to any one of paragraphs. 1-11, which is selected from the group consisting of: (1) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 42; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 41, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (2) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 46; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 45, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (3) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (4) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (5) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 54; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 53, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (6) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (7) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (8) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (9) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (10) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (11) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 34; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 33, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (12) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 34; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 33, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (13) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 28; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 27, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (14) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 28; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 27, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (15) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 30; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 29, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (16) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 30; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 29, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (17) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 58; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 57, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (18) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (19) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 60; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 59, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (20) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (21) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 64; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 63, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (22) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 48; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 47, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (23) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 48; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 47, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (24) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 41; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 42, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (25) the hybrid light chain includes SEQ ID NO: 58, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 45; the hybrid heavy chain includes SEQ ID NO: 57, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 46, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (26) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (27) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (28) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 53; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 54, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (29) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (30) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 67; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 68, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (31) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (32) the hybrid light chain includes SEQ ID NO: 68, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 67, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (33) the hybrid light chain includes SEQ ID NO: 42, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 41, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (34) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (35) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 27; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 28, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (36) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 27; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 28, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (37) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 29; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 30, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (38) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 29; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 30, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (39) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 57; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 58, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (40) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (41) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 59; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 60, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (42) the hybrid light chain includes SEQ ID NO: 38, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 37, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (43) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (44) the hybrid light chain includes SEQ ID NO: 46, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 63; the hybrid heavy chain includes SEQ ID NO: 45, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 64, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (45) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 47; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 48, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (46) the hybrid light chain includes SEQ ID NO: 50, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 47; the hybrid heavy chain includes SEQ ID NO: 49, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 48, SEQ ID NO: 74, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (47) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 149; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 148, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (48) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 147; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 146, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (49) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 149; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 148, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (50) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 147; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 146, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; (51) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 148; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 149, SEQ ID NO: 87, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (52) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 146; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 147, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 117 and SEQ ID NO: 120; (53) the hybrid light chain includes SEQ ID NO: 147, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 148; the hybrid heavy chain includes SEQ ID NO: 146, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 149, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120; or (54) the hybrid light chain includes SEQ ID NO: 149, SEQ ID NO: 104, SEQ ID NO: 85 and SEQ ID NO: 146; the hybrid heavy chain includes SEQ ID NO: 148, SEQ ID NO: 111, SEQ ID NO: 85, SEQ ID NO: 147, SEQ ID NO: 82, SEQ ID NO: 91, SEQ ID NO: 115 and SEQ ID NO: 120. 15. Pharmaceutical composition for the treatment of a tumor associated with the first epitope and the second epitope, including a bispecific antibody according to any one of paragraphs. 1-14. 16. The pharmaceutical composition according to claim 15, which is a dosage form for gastrointestinal administration or a dosage form for parenteral administration. 17. The pharmaceutical composition according to claim 16, wherein the dosage form of the pharmaceutical composition is an injection, including intravenous injection, intravenous infusion, subcutaneous injection, local injection, intramuscular injection, intratumoral injection, intraperitoneal injection, intracranial injection or intracavitary injection. 18. A conjugate that binds the first epitope and the second epitope, including a bispecific antibody according to any one of paragraphs. 1-14 and a therapeutic agent, prodrug, protein, virus, lipid, biological response modifier, PEG, hormone, oligonucleotide, diagnostic agent or cytotoxic agent. 19. A fusion protein that binds a first epitope and a second epitope, comprising a bispecific antibody according to any one of claims. 1-14 and a therapeutic agent, prodrug, protein, biological response modifier, hormone, diagnostic agent or cytotoxic agent. 20. The conjugate or fusion protein of claim 18 or 19, wherein the protein is an enzyme and the biological response modifier is an immunomodulator. 21. The conjugate or fusion protein of claim 18 or 19, wherein the diagnostic agent is selected from an ultrasound enhancing agent, a non-radioactive marker, a detectable marker. 22. The conjugate or fusion protein of claim 21, wherein the detectable marker is a chemiluminescent labeling compound or a fluorescent emitting metal. 23. The conjugate or fusion protein of claim 22, wherein the chemiluminescent labeling compound is selected from luminol, isoluminol, thermoluminescent acridinium ester, imidazole, acridinium salt and oxalate, and the fluorescent emitting metal is 152Eu or a lanthanide label. 24. The conjugate or fusion protein of claim 18 or 19, wherein the cytotoxic agent is selected from a drug and a toxin. 25. The conjugate or fusion protein of claim 18 or 19, wherein the protein is an enzyme and the biological response modifier is an immunomodulator. 26. A polynucleotide encoding a bispecific antibody that binds two different epitopes, according to any one of paragraphs. 1-14. 27. A cell expressing a bispecific antibody according to any one of paragraphs. 1-14, including the polynucleotide according to claim 26. 28. Use of a bispecific antibody according to any one of paragraphs. 1-14 or a conjugate or fusion protein according to any one of paragraphs. 18-25 for treating a tumor associated with the first epitope and the second epitope. 29. Use of a bispecific antibody according to any one of paragraphs. 1-14 or a conjugate or fusion protein according to any one of paragraphs. 18-25 to obtain a reagent or kit for detecting a tumor associated with a first epitope and a second epitope. 30. Use according to claim 28 or 29, wherein the tumor is cancer.

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