A simple two-step procedure for the preparation of the first component of human complement (C1) in its native form.
Native human C1 was purified from fresh human serum by affinity chromatography on protein A-bound Sepharose in the presence of 4-nitrophenyl-4-guanidinobenzoate hydrochloride (NPGB) taking advantage of the successive binding of IgG to protein A followed by C1 binding to IgG. After elution the C1 preparation contained IgG as a major contaminant as shown by SDS-PAGE. C1 was further purified by gel filtration. The yield of C1 was 12% and less than 4% of this C1 was activated during purification as assessed by a C4 consumption assay.
D002846:Chromatography, Affinity / Q000379:methods; D003172:Complement C1 / Q000302:isolation & purification*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000378:metabolism
D003172:Complement C1; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A