Inert format

Abstract

Described herein are, proteins comprising amino acid substitutions in at least one of a first and a second polypeptide chain. Furthermore, is described the uses and methods related to said proteins.

1 . A protein comprising a first polypeptide and a second polypeptide, wherein said first and second polypeptide each comprises at least a hinge region, a CH2 region and a CH3 region of an immunoglobulin heavy chain, wherein in at least one of said first and second polypeptide the amino acids in the positions corresponding to positions L234, L235 and D265 in a human IgG1 heavy chain are not L, L, and D, respectively. 
     
     
         2 . The protein according to  claim 1 , wherein in at least one of said first and second polypeptides the amino acid in the positions corresponding to positions L234, L235 and D265 in a human IgG1 heavy chain are not L, L, and D, respectively, and wherein the amino acids in the positions corresponding to positions N297 and P331 in a human IgG1 heavy chain are not Q and S, respectively. 
     
     
         3 . (canceled) 
     
     
         4 . The protein according to  claim 1 , wherein said first polypeptide and second polypeptide are a first heavy chain and a second heavy chain of an immunoglobulin, respectively. 
     
     
         5 . The protein according to  claim 1 , wherein said first polypeptide and second polypeptide further comprises a first binding region and a second binding region, respectively. 
     
     
         6 . (canceled) 
     
     
         7 . The protein according to  claim 5 , wherein at least one of said first and second binding regions bind CD3. 
     
     
         8 - 13 . (canceled) 
     
     
         14 . The protein according to  claim 1 , wherein in at least one of said first and second polypeptides, the amino acids corresponding to positions L234, L235 and D265 in a human IgG1 heavy chain are F, E, and A; or A, A, and A, respectively. 
     
     
         15 - 18 . (canceled) 
     
     
         19 . The protein according to  claim 5 , wherein said first binding region binds a different target than said second binding region. 
     
     
         20 . (canceled) 
     
     
         21 . The protein according to  claim 1 , wherein in said first polypeptide at least one of the amino acids in the positions corresponding to a position selected from the group consisting of T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and in said second polypeptide at least one of the amino acids in the positions corresponding to a position selected from the group consisting of; T366, L368, K370, D399, F405, Y407, and K409 in a human IgG1 heavy chain has been substituted, and wherein said substitutions of said first and said second polypeptides are not in the same positions. 
     
     
         22 . The protein according to  claim 21 , wherein the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first polypeptide, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second polypeptide, or vice versa. 
     
     
         23 . The protein according to  claim 1 , which is an antibody. 
     
     
         24 . The protein according to  claim 1 , wherein the protein is a bispecific antibody. 
     
     
         25 . The protein according to  claim 24 , wherein both said first and second polypeptide the amino acids in the positions corresponding to L234, L235 and D265 in a human IgG1 heavy chain are F, E, and A, respectively, said first binding region binds CD3, and said second binding region binds a cancer-specific target, and (i) wherein the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said first polypeptide, and the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said second polypeptide, or (ii) wherein the amino acid in the position corresponding to K409 in a human IgG1 heavy chain is R in said first polypeptide, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said second polypeptide. 
     
     
         26 - 28 . (canceled) 
     
     
         29 . A pharmaceutical composition comprising the protein of  claim 1  and a pharmaceutical acceptable carrier. 
     
     
         30 . A method of treatment of a disease comprising administering to a subject in need thereof an effective amount of the protein according to  claim 1 . 
     
     
         31 . (canceled) 
     
     
         32 . The method according to  claim 30 , wherein the disease is cancer, an infectious disease, or an autoimmune disease. 
     
     
         33 . A nucleic acid encoding (i) the first polypeptide, (ii) the second polypeptide, or (i) the first and second polypeptides of  claim 1 . 
     
     
         34 . A vector comprising the nucleic acid of  claim 33 . 
     
     
         35 . A host cell comprising the nucleic acid of  claim 33 . 
     
     
         36 . A method of detecting a target of interest in a sample comprising:
 (i) contacting the sample with the protein of  claim 1  under conditions that allow formation of a complex between the target of interest and protein, and   (ii) detecting the complex.   
     
     
         37 . The method of  claim 36 , wherein the sample is a biological sample.