Humanization of an antibody against human protein C and calcium-dependence involving framework residues.
A murine monoclonal antibody to the zymogen form of human protein C that blocks protein C activation by thrombin-thrombomodulin both in vitro and in vivo has been humanized using the consensus and resurfacing methods. While the binding of the parent murine antibody to protein C is calcium-dependent (1.5-2-fold decrease in binding without calcium), the humanized antibody exhibited a significant increase in its calcium-dependence (5-fold decrease in binding without calcium). Two exposed human framework residues in the variable light domain of the humanized antibody, aspartic acid L1 and glutamine L3, are responsible for the increase in calcium-dependence.
O'Connor
S J
SJ
Department of Immunology, Genentech, Inc., South San Francisco, CA 94080, USA.
Meng
Y G
YG
Rezaie
A R
AR
Presta
L G
LG
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D002118:Calcium / Q000378:metabolism*; D003001:Cloning, Molecular; D003198:Computer Simulation; D018076:DNA, Complementary; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011486:Protein C / Q000276:immunology*; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011486:Protein C; D002118:Calcium