Anti-cd3-binding domains and antibodies comprising them, and methods for their generation and use

Abstract

Anti-CD3 binding domains and antibodies comprising them, including multispecific antibodies, with, inter alia, desirable T-cell activation and (re)directed target cell killing potency and developability, profiles are provided, as well as methods for their identification, isolation, and generation, and methods for their preparation and use. Reagents for identifying, isolating, selecting, generating and characterizing CD3 binding domains and antibodies comprising them are also provided.

1 . An antibody or antigen-binding polypeptide comprising at least one of the following:
 a CDRH1 comprising an amino acid sequence of FNIKDYYMH (SEQ ID NO: 6720), YTFTSYTIH (SEQ ID NO: 6721), or FTFX 1 TYAMN (SEQ ID NO: 6722), wherein X 1  is any amino acid;   (ii) a CDRH2 comprising an amino acid sequence of WIDLENX 1 NTX 2 YDX 3 KFOG (SEQ ID NO: 6723), wherein X 1 , X 2 , and X 3  independently are any amino acid;   (iii) a CDRH3 comprising an amino acid sequence of X 1 QSYSX 2 RT (SEQ ID NO: 6724) or X 3 RDX 4 YGX 5 YFYDV (SEQ ID NO: 6725), wherein X 1 , X 2 , X 3 , X 4 , and X 5  are independently are any amino acid;   (iv) a CDRL1 comprising an amino acid sequence of X 1 KSSQX 2 LLX 3 X 4 RTGKX 5 YLA (SEQ ID NO: 6726), wherein X 1 , X 2 , X 3 , X 4 , and X 5  independently are any amino acid;   (v) a CDRL2 comprising an amino acid sequence of WASTRES (SEQ ID NO: 6727) or GTX1KRAP (SEQ ID NO: 6728), wherein X 1  is any amino acid; or   (vi) a CDRL3 comprising an amino acid sequence of KQSYSX 1 RT (SEQ ID NO: 6729), wherein X 1  is any amino acid.   
     
     
         2 . The antibody or antigen-binding polypeptide according to  claim 1 , wherein in said CDRH1 X 1  is N or D. 
     
     
         3 . (canceled) 
     
     
         4 . The antibody or antigen-binding polypeptide according to  claim 1 , wherein in said CDRH2:
 (i) X 1  is not G;   (ii) X 2  is V or I; and/or   (iii) X 3  is not G. 5-7. (Canceled)   
     
     
         8 . The antibody or antigen-binding polypeptide of  claim 1 , wherein in said CDRH3:
 (i) X 1  is K or T;   (ii) X 1  is T;   (iii) X 2  is R or L;   (iv) X 3  is A or G;   (v) X 4  is not G; and/or   (vi) X 5  is R, A, G, or L.   
     
     
         9 - 14 . (canceled) 
     
     
         15 . The antibody or antigen-binding polypeptide according to  claim 1 , wherein in said CDRL1:
 (i) X 1  is K or R;   (ii) X 2  is S or N;   (iii) X 3  is E or N;   (iv) X 4  is A or S; and/or   (v) X 5  is N or S.   
     
     
         16 - 20 . (canceled) 
     
     
         21 . The antibody or antigen-binding polypeptide according to  claim 1 , wherein in said CDRL2:
 X 1  is N or D.   
     
     
         22 . (canceled) 
     
     
         23 . The antibody or antigen-binding polypeptide according to  claim 1 , wherein in said CDRL3:
 (i) X 1  is R; or   (ii) X 1  is L or I.   
     
     
         24 . (canceled) 
     
     
         25 . An antibody or antigen binding polypeptide comprising at least two of a CDRH1, a CDRH2, and a CDRH3 according  claim 1 . 
     
     
         26 . An antibody or antigen-binding polypeptide comprising at least two of a CDRL1, a CDRL2, and a CDRL3 according to  claim 1 . 
     
     
         27 . An antibody or antigen-binding polypeptide comprising a CDRH1, a CDRH2, a CDRH3, and a CDRL1, a CDRL2, and a CDRL3 according to  claim 1 .