Anti-cd3 antibodies, anti-cd123 antibodies and bispecific antibodies specifically binding to cd3 and/or cd123

Abstract

The present invention concerns antibody-like binding protein specifically binding to CD3 and binding specifically to at least one further antigen, for example CD123. The present invention also concerns antibody-like binding protein specifically binding to CD123 and binding specifically to at least one further antigen. The invention further concerns anti-CD3 antibodies and anti-CD123 antibodies. The invention also relates to pharmaceutical compositions comprising the antibody-like binding protein, anti-CD3 antibodies or anti-CD123 antibodies of the invention, and their use to treat cancer. The invention further relates to isolated nucleic acids, vectors and host cells comprising a sequence encoding said antibody-like binding protein, anti-CD3 or anti-CD123 antibody and the use of said anti-CD123 antibody as a diagnostic tool.

1 : A method of treating a disease or disorder in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of an antibody-like binding protein that specifically binds to CD3ε and CD123 comprising two polypeptide chains that form two antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the formula [I]:
   V D1 -L 1 -V D2 -L 2 -C L   [I]
 
   
       and the second polypeptide chain comprises a structure represented by the formula [II]:
   V D3 -L 3 -V D4 -L 4 -C H1   [II]
 
 
       wherein:
 V D1  is a light chain variable domain of a first immuoglobulin; 
 V D2  is a light chain variable domain of a second immunoglobulin; 
 V D3  is a heavy chain variable domain of the second immunoglobulin; 
 V D4  is a heavy chain variable domain of the first immunoglobulin: 
 C L  is a light chain constant domain of an immunoglobulin; 
 C H1  is a C H1  heavy chain constant domain of an immunoglobulin; and 
 L 1 , L 2 , L 3 , and L 4  are amino acid linkers; 
 wherein the first polypeptide chain and the second polypeptide chain form a cross-over light chain-heavy chain pair; and 
 wherein V D1  comprises a CDR1-L comprising an amino acid sequence as set forth in SEQ ID NO: 378, a CDR2-L comprising an amino acid sequence of WAS, and a CDR3-L comprising an amino acid sequence as set forth in SEQ ID NO: 379; 
 V D2  comprises a CDR4-L comprising an amino acid sequence as set forth in SEQ ID NO: 142, a CDR5-L comprising an amino acid sequence of KVS, and a CDR6-L comprising an amino acid sequence as set forth in SEQ ID NO: 11: 
 V D3  comprises a CDR1-H comprising an amino acid sequence as set forth in SEQ ID NO: 6, a CDR2-H comprising an amino acid sequence as set forth in SEQ ID NO: 7, and a CDR3-H comprising an amino acid sequence as set forth in SEQ ID NO: 8; and 
 V D4  comprises a CDR4-H comprising an amino acid sequence as set forth in SEQ ID NO: 381, a CDR5-H comprising an amino acid sequence as set forth in SEQ ID NO: 384, and a CDR6-H comprising an amino acid sequence as set forth in SEQ ID NO: 382. 
 
     
     
         2 : The method of  claim 1 , wherein:
 L 1  comprises an amino acid sequence as set forth in SEQ ID NO: 389:   L 2  comprises an amino acid sequence as set forth in SEQ ID NO: 389; and   L 3  and L 4  are 0 amino acids in length.   
     
     
         3 : The method of  claim 1 , wherein:
 V D1  comprises an amino acid sequence as set forth in SEQ ID NO: 385:   V D2  comprises an amino acid sequence as set forth in SEQ ID NO: 141;   V D3  comprises an amino acid sequence as set forth in SEQ ID NO: 138;   V D4  comprises an amino acid sequence as set forth in SEQ ID NO: 383;   C L  comprises an amino acid sequence as set forth in SEQ ID NO: 310; and   C H1  comprises an amino acid sequence as set forth in SEQ ID NO: 313.   
     
     
         4 : The method of  claim 1 , wherein:
 (a) the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 388, or
 an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 388, which comprises a CDR1-L comprising an amino acid sequence as set forth in SEQ ID NO: 378, a CDR2-L comprising an amino acid sequence WAS, a CDR3-L comprising an amino acid sequence as set forth in SEQ ID NO: 379, a CDR4-L comprising an amino acid sequence as set forth in SEQ ID NO: 142, a CDR5-L comprising an amino acid sequence of KVS, and a CDR6-L comprising an amino acid sequence as set forth in SEQ ID NO: 11; and 
   (b) the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 390, or
 an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 390, which comprises a CDR1-H comprising an amino acid sequence as set forth in SEQ ID NO: 6, a CDR2-H comprising an amino acid sequence as set forth in SEQ ID NO: 7, a CDR3-H comprising an amino acid sequence as set forth in SEQ ID NO: 8, a CDR4-H comprising an amino acid sequence as set forth in SEQ ID NO: 381, a CDR5-H comprising an amino acid sequence as set forth in SEQ ID NO: 384, and a CDR6-H comprising an amino acid sequence as set forth in SEQ ID NO: 382. 
   
     
     
         5 : The method of  claim 1 , wherein the second polypeptide chain further comprises a Fc domain (“Fc”). 
     
     
         6 : The method of  claim 1 , wherein the first polypeptide chain further comprises a Fc domain (“Fc”). 
     
     
         7 : A method of treating a disease or disorder in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of an antibody-like binding protein that specifically binds to CD3ε and CD123 comprising two polypeptide chains that form two antigen-binding sites, wherein the first polypeptide chain comprises a structure represented by the formula [IV]:
   V D1 -L 1 -V D2 -L 2 -C L -L 5 -Fc 2   [IV]
 
   
       and the second polypeptide chain comprises a structure represented by the formula [III]:
   V D3 -L 3 -V D4 -L 4 -C H1 -Fc  [III]
 
 
       wherein:
 V D1  is a light chain variable domain of a first immunoglobulin; 
 V D2  is a light chain variable domain of a second immuoglobulin; 
 V D3  is a heavy chain variable domain of the second immunoglobulin; 
 V D4  is a heavy chain variable domain of the first immunoglobulin; 
 C L  is a light chain constant domain of an immunoglobulin; 
 C H1  is a C H1  heavy chain constant domain of an immunoglobulin; 
 Fc is the immunoglobulin hinge region and CH 2 , CH 3  immunoglobulin heavy chain constant domains of a first immunoglobulin; 
 Fc 2  is the immunoglobulin hinge region and CH 2 , CH 3  immunoglobulin heavy chain constant domains of a second immunoglobulin; and 
 L 1 , L 2 , L 3 , L 4 , and L 5  are amino acid linkers; 
 wherein the first polypeptide chain of formula [IV] and the second polypeptide chain of formula [III] form a cross-over light chain-heavy chain pair; and 
 wherein V D1  comprises a CDR1-L comprising an amino acid sequence as set forth in SEQ ID NO: 378, a CDR2-L comprising an amino acid sequence of WAS, and a CDR3-L comprising an amino acid sequence as set forth in SEQ ID NO: 379; 
 V D2  comprises a CDR4-L comprising an amino acid sequence as set forth in SEQ ID NO: 142, a CDR5-L comprising an amino acid sequence of KVS, and a CDR6-L comprising an amino acid sequence as set forth in SEQ ID NO: 11; 
 V D3  comprises a CDR1-H comprising an amino acid sequence as set forth in SEQ ID NO: 6, a CDR2-H comprising an amino acid sequence as set forth in SEQ ID NO: 7, and a CDR3-H comprising an amino acid sequence as set forth in SEQ ID NO: 8; and 
 V D4  comprises a CDR4-H comprising an amino acid sequence as set forth in SEQ ID NO: 381, a CDR5-H comprising an amino acid sequence as set forth in SEQ ID NO: 384, and a CDR6-H comprising an amino acid sequence as set forth in SEQ ID NO: 382. 
 
     
     
         8 : The method of  claim 7 , wherein:
 L 1  comprises an amino acid sequence as set forth in SEQ ID NO: 389:   L 2  comprises an amino acid sequence as set forth in SEQ ID NO: 389; and   L 3 , L 4 , and L 5  are 0 amino acids in length.   
     
     
         9 : The method of  claim 7 , wherein:
 Fc comprises an amino acid sequence as set forth in SEQ ID NO: 394; and   Fc 2  comprises an amino acid sequence as set forth in SEQ ID NO: 392.   
     
     
         10 : The method of  claim 7 , wherein:
 V D1  comprises an amino acid sequence as set forth in SEQ ID NO: 385:   V D2  comprises an amino acid sequence as set forth in SEQ ID NO: 141;   V D3  comprises an amino acid sequence as set forth in SEQ ID NO: 138;   V D4  comprises an amino acid sequence as set forth in SEQ ID NO: 383;   C L  comprises an amino acid sequence as set forth in SEQ ID NO: 310; and   C H1  comprises an amino acid sequence as set forth in SEQ ID NO: 313.   
     
     
         11 : The method of  claim 7 , wherein:
 (a) the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 391, or   an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 391, which comprises a CDR1-L comprising an amino acid sequence as set forth in SEQ ID NO: 378, a CDR2-L comprising an amino acid sequence WAS, a CDR3-L comprising an amino acid sequence as set forth in SEQ ID NO: 379, a CDR4-L comprising an amino acid sequence as set forth in SEQ ID NO: 142, a CDR5-L comprising an amino acid sequence of KVS, and a CDR6-L comprising an amino acid sequence as set forth in SEQ ID NO: 11; and   (b) the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 393, or   an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 393, which comprises a CDR1-H comprising an amino acid sequence as set forth in SEQ ID NO: 6, a CDR2-H comprising an amino acid sequence as set forth in SEQ ID NO: 7, a CDR3-H comprising an amino acid sequence as set forth in SEQ ID NO: 8, a CDR4-H comprising an amino acid sequence as set forth in SEQ ID NO: 381, a CDR5-H comprising an amino acid sequence as set forth in SEQ ID NO: 384, and a CDR6-H comprising an amino acid sequence as set forth in SEQ ID NO: 382.   
     
     
         12 : A method of treating a disease or disorder in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of an antibody-like binding protein that binds specifically to CD3ε and CD123 comprising two polypeptide chains that form two antigen-binding sites, wherein a first polypeptide has a structure represented by the formula [I]:
   V D1 -L 1 -V D2 -L 2 -C L   [I]
 
   
       and a second polypeptide has a structure represented by the formula [II]:
   V D3 -L 3 -V D4 -L 4 -C H1   [II]
 
 
       wherein:
 V D1  is a light chain variable domain of a first immunoglobulin; 
 V D2  is a light chain variable domain of a second immuoglobulin; 
 V D3  is a heavy chain variable domain of the second immunoglobulin; 
 V D4  is a heavy chain variable domain of the first immunoglobulin; 
 C L  is a light chain constant domain of an immunoglobulin; 
 C H1  is a C H1  heavy chain constant domain of an immunoglobulin; 
 L 1 , L 2 , L 3 , and L 4  are amino acid linkers; and 
 wherein the first polypeptide and the second polypeptide form a cross-over light chain-heavy chain pair; and 
 wherein V D1  consists of the amino acid sequence as set forth in SEQ ID NO: 385, or an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 385, which comprises a CDR1-L of sequence SEQ ID NO: 378, a CDR2-L of sequence WAS, and a CDR3-L of sequence SEQ ID NO: 379: 
 V D2  consists of the amino acid sequence as set forth in SEQ ID NO: 141, or an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 141, which comprises a CDR1-L of sequence SEQ ID NO: 142, a CDR2-L of sequence KVS, and a CDR3-L of sequence SEQ ID NO: 11; 
 V D3  consists of the amino acid sequence as set forth in SEQ ID NO: 138, or an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 138, which comprises a CDR1-H of sequence SEQ ID NO: 6, a CDR2-H of sequence SEQ ID NO: 7, and a CDR3-H of sequence SEQ ID NO: 8; and 
 V D4  consists of the amino acid sequence as set forth in SEQ ID NO: 383, or an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 383, which comprises a CDR1-H of sequence SEQ ID NO: 381, a CDR2-H of sequence SEQ ID NO: 384, and a CDR3-H of sequence SEQ ID NO: 382. 
 
     
     
         13 : The method of  claim 12 , wherein:
 (a) the first polypeptide consists of the amino acid sequence as set forth in SEQ ID NO: 388, which comprises V D1  of amino acid sequence SEQ ID NO: 385, L 1  of amino acid sequence SEQ ID NO: 389, V D2  of amino acid sequence SEQ ID NO: 141, L 2  of amino acid sequence SEQ ID NO: 389, and C L  of amino acid sequence SEQ ID NO: 310: or   an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 388, wherein the three CDRs of amino acid sequences SEQ ID NO: 378, WAS, and SEQ ID NO: 379 of V D1 , and the three CDRs of amino acid sequences SEQ ID NO: 142, KVS, and SEQ ID NO: 11 of V D2  are unaltered; and   (b) the second polypeptide consists of the amino acid sequence as set forth in SEQ ID NO: 390, which comprises V D3  of amino acid sequence SEQ ID NO: 138, L 3  is 0 amino acids, V D4  of amino acid sequence SEQ ID NO: 383, L 4  is 0 amino acids, and C H1  of amino acid sequence SEQ ID NO: 313, or   an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 390, wherein the three CDRs of amino acid sequences SEQ ID NO: 381, SEQ ID NO: 384, and SEQ ID NO: 382 of V D4 , and the three CDRs of amino acid sequences SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8 of V D3  are unaltered.   
     
     
         14 : The method of  claim 12 , wherein the polypeptide of formula [II] further comprises a Fc domain and has a structure represented by formula [III]:
   V D3 -L 3 -V D4 -L 4 -C H1 -Fc  [III]
   wherein the polypeptide of formula [I] further comprises a Fc domain (Fc 2 ) and has a structure represented by formula [IV]:
   V D1 -L 1 -V D2 -L 2 -C L -L 5 -Fc 2   [IV]
 
   wherein:
 Fc is the immunoglobulin hinge region and CH 2 , CH 3  immunoglobulin heavy chain constant domains of an immunoglobulin, 
 Fc 2  is the immunoglobulin hinge region and CH 2 , CH 3  immunoglobulin heavy chain constant domains of an immunoglobulin, and 
 L 5  is an amino acid linker; and 
   wherein:
 (a) the polypeptide according to formula [IV] consists of the amino acid sequence as set forth in SEQ ID NO: 391, which comprises V D1  of amino acid sequence SEQ ID NO: 385, L 1  of amino acid sequence SEQ ID NO: 389, V D1  of amino acid sequence SEQ ID NO: 141, L 2  of amino acid sequence SEQ ID NO: 389, C L  of amino acid sequence SEQ ID NO: 310, L 5  which is 0 amino acids, and Fc 2  of amino acid sequence SEQ ID NO: 392; or 
 an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 391, wherein the three CDRs of amino acid sequences SEQ ID NO: 378, WAS, and SEQ ID NO: 379 of V D1 , and the three CDRs of amino acid sequences SEQ ID NO: 142, KVS, and SEQ ID NO: 11 of V D2  are unaltered; and 
 (b) the polypeptide according to formula [III] consists of the amino acid sequence as set forth in SEQ ID NO: 393, which comprises V D3  of amino acid sequence SEQ ID NO: 138, L 3  is 0 amino acids, V D4  of amino acid sequence SEQ ID NO: 383, L 4  is 0 amino acids, C H1  of amino acid sequence SEQ ID NO: 313, and Fc of amino acid sequence SEQ ID NO: 394: or 
 an amino acid sequence that is at least 85% identical to the amino acid sequence as set forth in SEQ ID NO: 393, wherein the three CDRs of amino acid sequences SEQ ID NO: 381, SEQ ID NO: 384, and SEQ ID NO: 382 of V D4 , and the three CDRs of amino acid sequences SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8 of V D3  are unaltered; and 
   wherein the polypeptide of formula [IV] and the polypeptide of formula [III] form a cross-over light chain-heavy chain pair.   
     
     
         15 : A method of treating a disease or disorder in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of an antibody-like binding protein comprising two polypeptide chains that form two antigen-binding sites, wherein:   a first polypeptide chain comprises:
 a CDR1-L comprising an amino acid sequence as set forth in SEQ ID NO: 378; 
 a CDR2-L comprising an amino acid sequence WAS: 
 a CDR3-L comprising an amino acid sequence as set forth in SEQ ID NO: 379; 
 a CDR4-L comprising an amino acid sequence as set forth in SEQ ID NO: 142: 
 a CDR5-L comprising an amino acid sequence of KVS; and 
 a CDR6-L comprising an amino acid sequence as set forth in SEQ ID NO: 11: and 
   wherein a second polypeptide chain comprises:
 a CDR1-H comprising an amino acid sequence as set forth in SEQ ID NO: 6: 
 a CDR2-H comprising an amino acid sequence as set forth in SEQ ID NO: 7; 
 a CDR3-H comprising an amino acid sequence as set forth in SEQ ID NO: 8; 
 a CDR4-H comprising an amino acid sequence as set forth in SEQ ID NO: 381; 
 a CDR5-H comprising an amino acid sequence as set forth in SEQ ID NO: 384; and 
 a CDR6-H comprising an amino acid sequence as set forth in SEQ ID NO: 382. 
   
     
     
         16 : The method of  claim 1 , wherein the disease or disorder is cancer or a pathological immune response. 
     
     
         17 : The method of  claim 16 , wherein the cancer is a hematological cancer associated with CD123 expression. 
     
     
         18 : The method of  claim 17 , wherein the hematological cancer is a leukemia, a malignant lymphoproliferative condition, a blastic plasmacytoid dendritic cell neoplasm (BPDCN), systemic mastocytosis, or a lymphoma. 
     
     
         19 : The method of  claim 18 , wherein the leukemia is acute myelogenous leukemia, chronic myelogenous leukemia, acute lymphoid leukemia, chronic lymphoid leukemia, hairy cell leukemia, or myelodysplasia syndrome. 
     
     
         20 : The method of  claim 19 , wherein the leukemia is acute myelogenous leukemia. 
     
     
         21 : The method of  claim 18 , wherein the lymphoma is multiple myeloma, non-Hodgkin's lymphoma, Burkitt's lymphoma, small cell-follicular lymphoma, or large cell-follicular lymphoma. 
     
     
         22 : The method of  claim 16 , wherein the pathological immune response comprises an autoimmune disease, a transplantation-related disease, or an inflammation-associated disease. 
     
     
         23 : The method of  claim 22 , wherein the autoimmune disease is Crohn's disease, ulcerative colitis, or type I diabetes. 
     
     
         24 : The method of  claim 22 , wherein the transplantation-related disease is graft-versus-host disease (GVHD). 
     
     
         25 : The method of  claim 1 , wherein the antibody-like binding protein is comprised within a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier. 
     
     
         26 : The method of  claim 1 , wherein the antibody-like binding protein induces T-cell mediated toxicity of CD123 positive tumor cells within the subject. 
     
     
         27 : The method of  claim 1 , wherein the antibody-like binding protein induces an immunosuppressive effect in the subject. 
     
     
         28 : A method of treating a disease or disorder in a subject in need thereof, comprising:
 administering to the subject a therapeutically effective amount of an antibody-like binding protein that specifically binds to CD3ε and CD123 comprising two polypeptide chains that form two antigen-binding sites,   wherein a first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 388 and a second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 390 or the first polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 391 and the second polypeptide chain comprises an amino acid sequence as set forth in SEQ ID NO: 393.   
     
     
         29 : An isolated antibody that binds to the extracellular domain of human CD3ε protein comprising:
 a) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence GFX 1 X 2 X 3 X 4 AW (SEQ ID NO: 331), wherein X 1  is T or S, X 2  is F or V, X 3  is S or T and X 4  is N, K, L, or Y, or any combination thereof; a CDR2-H comprising amino acid sequence IKX 1 X 2 X 3 NX 4 YX 5 T (SEQ ID NO: 332), wherein X 1  is A or D, X 2  is K or R, X 3  is S or A, X 4  is N or S, and X 5  is A or E, or any combination thereof; and a CDR3-H comprising amino acid sequence TWRHYYSSHTMDA (SEQ ID NO: 69), or RALTYYGYKRDAMDG (SEQ ID NO: 129), or RX 1 X 2 X 3 YX 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 DX 12  (SEQ ID NO: 333), wherein X 1  is Y, G, or A, X 2  is V, T, or L, X 3  is H, N, Y, or Q, X 4  is G, R, or A, X 5  is F or V or no amino acid, X 6  is R or no amino acid, X 7  is F, S, or I or no amino acid, X 8  is F, L, N, M, Y, S, A, or G, X 9  is Y, A, K, S, N, T, F, or L, X 10  is A, P, G, or T, X 11  is M, L, F, or S, and X 12  is A, V, or Y, or any combination thereof; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence QX 1 LX 2 HX 3 NGX 4 TY (SEQ ID NO: 334) wherein X 1  is R or S, X 2  is V or E, X 3  is N, D, or T, and X 4  is N or Y, or any combination thereof; and a CDR2-L comprising amino acid sequence KVS; and a CDR3-L comprising an amino acid sequence GQGX 1 X 2 YPFT (SEQ ID NO: 335) wherein X 1  is T, A or S and X 2  is H, E or Q, or any combination thereof; or 
 b) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 30 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 30 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 31 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 31 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 32 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 32 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 34 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 34 by one amino acid substitution; a CDR2-L comprising amino acid sequence RDD or an amino acid sequence differing from the amino acid sequence RDD by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 35 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 35 by one amino acid substitution; or 
 c) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 50 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 50 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 51 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 51 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 52 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 52 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 54 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 54 by one amino acid substitution; a CDR2-L comprising amino acid sequence NAN or an amino acid sequence differing from the amino acid sequence NAN by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 55 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 55 by one amino acid substitution; or 
 d) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 90 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 90 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 91 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 91 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 32 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 32 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 93 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 93 by one amino acid substitution; a CDR2-L comprising amino acid sequence GAS or an amino acid sequence differing from the amino acid sequence GAS by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 94 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 94 by one amino acid substitution; or 
 e) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 96 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 96 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 97 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 97 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 98 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 98 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 100 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 100 by one amino acid substitution, a CDR2-L comprising amino acid sequence NTN or an amino acid sequence differing from the amino acid sequence NTN by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 101 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 101 by one amino acid substitution; or 
 f) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 103 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 103 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 104 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 104 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 105 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 105 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: by one amino acid substitution; a CDR2-L comprising amino acid sequence KVS or an amino acid sequence differing from the amino acid sequence KVS by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 11 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 11 by one amino acid substitution; or 
 g) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 116 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 116 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 117 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 117 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 118 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 118 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 100 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 100 by one amino acid substitution; a CDR2-L comprising amino acid sequence VTN or an amino acid sequence differing from the amino acid sequence VTN by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 120 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 120 by one amino acid substitution; or 
 h) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 122 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 122 by one amino acid substitution; a CDR2-H comprising amino acid sequence SEQ ID NO: 123 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 123 by one or more amino acid substitutions; and a CDR3-H comprising amino acid sequence SEQ ID NO: 124 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 124 by one amino acid substitution; and 
 a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 126 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 126 by one amino acid substitution; a CDR2-L comprising amino acid sequence RDD or an amino acid sequence differing from the amino acid sequence RDD by one amino acid substitution; and a CDR3-L comprising amino acid sequence SEQ ID NO: 127 or an amino acid sequence differing from the amino acid sequence SEQ ID NO: 127 by one amino acid substitution. 
 
     
     
         30 : The isolated antibody according to  claim 29 , comprising:
 a) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 6, a CDR2-H comprising amino acid sequence SEQ ID NO: 7, and a CDR3-H of amino acid sequence SEQ ID NO: 8; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10 or SEQ ID NO: 142, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   b) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 14, and a CDR3-H comprising amino acid sequence SEQ ID NO: 15; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 17 or SEQ ID NO: 184, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   c) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 19, and a CDR3-H comprising amino acid sequence SEQ ID NO: 20; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 22, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   d) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 24, a CDR2-H comprising amino acid sequence SEQ ID NO: 19, and a CDR3-H comprising amino acid sequence SEQ ID NO: 25; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 27; a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 28; or   e) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 30, a CDR2-H comprising amino acid sequence SEQ ID NO: 31, and a CDR3-H comprising amino acid sequence SEQ ID NO: 32: and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 34, a CDR2-L comprising amino acid sequence ROD, and a CDR3-L comprising amino acid sequence SEQ ID NO: 35; or   f) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 38; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 28; or   g) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 41; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 17, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   h) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 44; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   i) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 47; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   j) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 50, a CDR2-H comprising amino acid sequence SEQ ID NO: 51, and a CDR3-H comprising amino acid sequence SEQ ID NO: 52; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 54, a CDR2-L comprising amino acid sequence NAN, and a CDR3-L comprising amino acid sequence SEQ ID NO: 55; or   k) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 57, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 58; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 28; or   l) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 61; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   m) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 64, a CDR2-H comprising amino acid sequence SEQ ID NO: 65, and a CDR3-H comprising amino acid sequence SEQ ID NO: 47: and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 67, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 28; or   n) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 69; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 71; or   o) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 84; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 17, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   p) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 75, a CDR2-H comprising amino acid sequence SEQ ID NO: 76, and a CDR3-H comprising amino acid sequence SEQ ID NO: 77; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   q) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 80, a CDR2-H comprising amino acid sequence SEQ ID NO: 76, and a CDR3-H comprising amino acid sequence SEQ ID NO: 81; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   r) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 84; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   s) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 47: and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 88; or   t) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 90, a CDR2-H comprising amino acid sequence SEQ ID NO: 91, and a CDR3-H comprising amino acid sequence SEQ ID NO: 32; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 93, a CDR2-L comprising amino acid sequence GAS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 94; or   u) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 96, a CDR2-H comprising amino acid sequence SEQ ID NO: 97, and a CDR3-H comprising amino acid sequence SEQ ID NO: 98; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 100, a CDR2-L comprising amino acid sequence NTN, and a CDR3-L comprising amino acid sequence SEQ ID NO: 101; or   v) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 103, a CDR2-H comprising amino acid sequence SEQ ID NO: 104, and a CDR3-H comprising amino acid sequence SEQ ID NO: 105; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   w) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 80, a CDR2-H comprising amino acid sequence SEQ ID NO: 19, and a CDR3-H comprising amino acid sequence SEQ ID NO: 108; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   x) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 37, and a CDR3-H comprising amino acid sequence SEQ ID NO: 111; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 113, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 114; or   y) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 116, a CDR2-H comprising amino acid sequence SEQ ID NO: 117, and a CDR3-H comprising amino acid sequence SEQ ID NO: 118; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 100, a CDR2-L comprising amino acid sequence VTN, and a CDR3-L comprising amino acid sequence SEQ ID NO: 120; or   z) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 122, a CDR2-H comprising amino acid sequence SEQ ID NO: 123, and a CDR3-H comprising amino acid sequence SEQ ID NO: 124; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 126, a CDR2-L comprising amino acid sequence ROD, and a CDR3-L comprising amino acid sequence SEQ ID NO: 127; or   aa) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 13, a CDR2-H comprising amino acid sequence SEQ ID NO: 19, and a CDR3-H comprising amino acid sequence SEQ ID NO: 129; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 10, a CDR2-L comprising amino acid sequence KVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 11; or   bb) a heavy chain variable domain comprising a CDR1-H comprising amino acid sequence SEQ ID NO: 103, a CDR2-H comprising amino acid sequence SEQ ID NO: 104, and a CDR3-H comprising amino acid sequence SEQ ID NO: 105; and a light chain variable domain comprising a CDR1-L comprising amino acid sequence SEQ ID NO: 133, a CDR2-L comprising amino acid sequence LVS, and a CDR3-L comprising amino acid sequence SEQ ID NO: 134.   
     
     
         31 : An antibody-like binding protein that binds specifically to human CD3ε comprising two polypeptide chains that form two antigen-binding sites, wherein a first polypeptide has a stricture represented by the formula [I]:
   VD-L 1 -V D1 -L 2 -C L   [I]
 
 and a second polypeptide has a structure represented by the formula [II]:
   V D3 -L 3 -V D4 -L 4 -C H1   [II]
 
 
 wherein:
 V D1  is a heavy or light chain variable domain of a first immunoglobulin; 
 V D2  is a heavy or light chain variable domain of a second immuoglobulin; 
 V D3  is a heavy or light chain variable domain of the second immunoglobulin; 
 V D4  is a heavy or light chain variable domain of the first immunoglobulin; 
 C L  is a light chain constant domain of an immunoglobulin; 
 C H1  is a C H1  heavy chain constant domain of an immunoglobulin; 
 L 1 , L 2 , L 3 , and L 4  are amino acid linkers; 
 wherein the first polypeptide and the second polypeptide form a cross-over light chain-heavy chain pair; and 
 wherein V D1  and V D4 , or V D2  and V D3  comprise a heavy chain variable domain and a light chain variable domain of an antibody according to  claim 29 . 
 
 
     
     
         32 : The antibody-like binding protein of  claim 31 , wherein at least one of polypeptide [III] and polypeptide [IV] further comprises an Fc domain.