Preparation of monoclonal antibodies to C3b by immunization with C3b(i)-sepharose.
We have prepared and characterized four monoclonal antibodies (MAbs) to human C3b of high specificity and affinity. Our procedure did not require a purified source of C3b for immunization. Instead, C3b and C3bi were deposited on Sepharose 4B via the alternative pathway of complement activation in normal human serum, and this C3b(i)-Sepharose served as the immunogen. C3b(i)-Sepharose was also prepared from a number of primate and non-primate sources, and this allowed us to demonstrate that the anti-human C3b MAbs cross-reacted with primate-derived C3b, but not with C3b from non-primates. The procedures we have developed may be useful in the further investigation of species-specific C3 fragment-binding proteins from both primate and non-primate sources.
Tosic
L
L
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908.
Sutherland
W M
WM
Kurek
J
J
Edberg
J C
JC
Taylor
R P
RP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens; D003176:Complement C3 / Q000276:immunology; D003179:Complement C3b / Q000276:immunology*; D015933:Complement C3d; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D012685:Sepharose
D000911:Antibodies, Monoclonal; D000941:Antigens; D003176:Complement C3; D003179:Complement C3b; D015933:Complement C3d; D012685:Sepharose