A general method for affinity purification of complement component C3b using factor H-sepharose.
Complement component C3b has been purified from human, rabbit and bovine serum by affinity chromatography on human factor H-Sepharose after preliminary fractionation by poly(ethylene glycol) and DEAE-Sepharose. The yields are high (35--40%) and the whole process is rapid (3 days). Binding of C3b to factor H-Sepharose is equimolar, has a sharp optimum pH at 7.6 and is quite sensitive to ionic strength.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange; D003179:Complement C3b / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D011485:Protein Binding; D011817:Rabbits; D012685:Sepharose
D003179:Complement C3b; D012685:Sepharose