The first component of complement. I. Purification and properties of native C1.
The first component of complement has been purified by using affinity chromatography on Sepharose-bound IgG. Unlike earlier procedures that yield the activated form of C1, in this method C1 is maintained in the native form by the protease inhibitor p-nitrophenyl, p'-guanidinobenzoate (NPGB). The procedure requires only two steps and yields pure C1 as judged both by SDS-PAGE analysis and by effective molecule calculations. The yields have varied from 30 to 50% in over 50 preparations. The functional properties of the purified native C1 correspond to those of C1 in serum. The dose-response activity profile is nonlinear, but becomes linear when C1 IS ALLOWED TO SELF-ACTIVATE. From SDS-PAGE analysis of the self-activated C1, all the C1r and C1s subcomponents are converted to the activated split products, indicating that all C1 molecules are biologically active. The recovery of C1 activity is dependent on the use of a heterologous source for the IgG on the affinity absorbant. The conditions of binding and elution from the Sepharose-IgG column are critical, indicating that immunoglobulin-bound C1 is rapidly inactivated under physiologic conditions by serum inactivators. The activation of the purified C1 in fluid phase has been explored both in the presence and absence of C1-inhibitor.
D001565:Benzoates / Q000494:pharmacology; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003167:Complement Activation; D003172:Complement C1 / Q000302:isolation & purification*; D003174:Complement C1 Inactivator Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D006146:Guanidines / Q000494:pharmacology; D006461:Hemolysis; D006801:Humans; D007074:Immunoglobulin G; D008970:Molecular Weight; D012685:Sepharose; D013395:Sucrose / Q000494:pharmacology
D001565:Benzoates; D003172:Complement C1; D003174:Complement C1 Inactivator Proteins; D006146:Guanidines; D007074:Immunoglobulin G; C002238:4-nitrophenyl 4'-guanidinobenzoate; D013395:Sucrose; D012685:Sepharose