Antibody Constant Region Variant

Abstract

By altering amino acid sequences, the present inventors successfully produced constant regions that can confer antibodies with particularly favorable properties for pharmaceutical agents. When used to produce antibodies, the altered constant regions produced according to the present invention significantly reduce heterogeneity. Specifically, the antibody homogeneity can be achieved by using antibody heavy chain and light chain constant regions introduced with alterations provided by the present invention. More specifically, the alterations can prevent the loss of homogeneity of antibody molecules due to disulfide bond differences in the heavy chain. Furthermore, in a preferred embodiment, the present invention can improve antibody pharmacokinetics as well as prevent the loss of homogeneity due to C-terminal deletion in antibody constant region.

1 . An antibody constant region comprising an amino acid sequence in which Cys at position 14 (position 131 in the EU numbering), Arg at position 16 (position 133 in the EU numbering), Cys at position 103 (position 220 in the EU numbering), Glu at position 20 (position 137 in the EU numbering), Ser at position 21 (position 138 in the EU numbering), His at position 147 (position 268 in the EU numbering), Arg at position 234 (position 355 in the EU numbering), and Gln at position 298 (position 419 in the EU numbering) in the amino acid sequence of SEQ ID NO: 24 (IgG2 constant region) are substituted with other amino acids and, optionally, wherein Gly at position 325 (position 446 in the EU numbering) and Lys at position 326 (position 447 in the EU numbering) are deleted. 
     
     
         2 . The antibody constant region of  claim 1 , wherein Ser is substituted for Cys at position 14, Lys is substituted for Arg at position 16, Ser is substituted for Cys at position 103, Gly is substituted for Glu at position 20, Gly is substituted for Ser at position 21, Gln is substituted for His at position 147, Gln is substituted for Arg at position 234, and Glu is substituted for Gln at position 298. 
     
     
         3 . The antibody constant region of  claim 1 , which comprises an amino acid sequence additionally having deletion of Gly at position 325 (position 446 in the EU numbering) and Lys at position 326 (position 447 in the EU numbering). 
     
     
         4 . An antibody constant region comprising an amino acid sequence in which Cys at position 14 (position 131 in the EU numbering), Arg at position 16 (position 133 in the EU numbering), Cys at position 103 (position 220 in the EU numbering), Glu at position 20 (position 137 in the EU numbering), Ser at position 21 (position 138 in the EU numbering), His at position 147 (position 268 in the EU numbering), Arg at position 234 (position 355 in the EU numbering), Gln at position 298 (position 419 in the EU numbering), Ala at position 209 (position 330 in the EU numbering), Pro at position 210 (position 331 in the EU numbering), and Thr at position 218 (position 339 in the EU numbering) in the amino acid sequence of SEQ ID NO: 24 (IgG2 constant region) are substituted with other amino acids and, optionally, wherein Gly at position 325 (position 446 in the EU numbering) and Lys at position 326 (position 447 in the EU numbering) are deleted. 
     
     
         5 . The antibody constant region of  claim 4 , wherein Ser is substituted for Cys at position 14, Lys is substituted for Arg at position 16, Ser is substituted for Cys at position 103, Gly is substituted for Glu at position 20, Gly is substituted for Ser at position 21, Gln is substituted for His at position 147, Gln is substituted for Arg at position 234, Glu is substituted for Gln at position 298, Ser is substituted for Ala at position 209, Ser is substituted for Pro at position 210, and Ala is substituted for Thr at position 218. 
     
     
         6 . The antibody constant region of  claim 4 , which also comprises an amino acid sequence in which Gly at position 325 (position 446 in the EU numbering) and Lys at position 326 (position 447 in the EU numbering) are deleted. 
     
     
         7 . An antibody comprising the constant region of  claim 1 . 
     
     
         8 . A pharmaceutical composition comprising the antibody of  claim 7 . 
     
     
         9 . A human κ chain constant region comprising at least one Cys at positions 102 to 106. 
     
     
         10 . A human κ chain constant region which does not comprise Cys at position 107. 
     
     
         11 . A human κ chain constant region which comprises at least one Cys at positions 102 to 106 but does not comprise Cys at position 107. 
     
     
         12 . The human κ chain constant region of  claim 9 , in which at least one amino acid at positions 1 to 106 in the amino acid sequence of SEQ ID NO: 32 is deleted. 
     
     
         13 . The human κ chain constant region of  claim 12 , in which at least one amino acid at positions 102 to 106 is deleted. 
     
     
         14 . The human κ chain constant region of  claim 13 , in which the amino acid at position 105 is deleted. 
     
     
         15 . The human κ chain constant region of  claim 13 , in which the amino acid at position 106 is deleted. 
     
     
         16 . The human κ chain constant region of  claim 9 , in which at least one amino acid at position 102 to 106 is substituted with Cys. 
     
     
         17 . The human κ chain constant region of  claim 9 , in which at least one amino acid at positions 102 to 106 is substituted with Cys, and Cys at position 107 is deleted or substituted with another amino acid. 
     
     
         18 . An antibody comprising the human κ chain constant region of  claim 9 . 
     
     
         19 . A pharmaceutical composition comprising the antibody of  claim 18 . 
     
     
         20 . An antibody comprising a heavy chain constant region comprising:
 an amino acid sequence in which Cys at position 14 (position 131 in the EU numbering), Arg at position 16 (position 133 in the EU numbering), Cys at position 103 (position 220 in the EU numbering), Glu at position 20 (position 137 in the EU numbering), Ser at position 21 (position 138 in the EU numbering), His at position 147 (position 268 in the EU numbering), Arg at position 234 (position 355 in the EU numbering), and Gln at position 298 (position 419 in the EU numbering) in the amino acid sequence of SEQ ID NO: 24 (IgG2 constant region) are substituted with other amino acids and, optionally, wherein Gly at position 325 (position 446 in the EU numbering) and Lys at position 326 (position 447 in the EU numbering) are deleted;   and a light chain constant region selected from:   (1) a human κ chain constant region comprising at least one Cys at positions 102 to 106;   (2) a human κ chain constant region which does not comprise Cys at position 107; and   (3) A human κ chain constant region which comprises at least one Cys at positions 102 to 106 but does not comprise Cys at position 107.   
     
     
         21 . A pharmaceutical composition comprising the antibody of  claim 20 .