An efficient one-step method for isolating immune complexes from whole serum using a monoclonal anti-C3g affinity immunosorbent

Abstract

A simple and efficient method of extracting complement-fixing immune complexes (IC) from whole serum and recovering them has been developed. 125I-labelled, in vitro prepared IC in whole serum were incubated with Sepharose 4B covalently linked to monoclonal anti-C3g or anti-C1q and the binding and recovery of IC was monitored by radioactivity. The anti-C3g immunosorbent bound 45% and 76% of HAGG and BSA-anti-BSA IC respectively, all of which were recovered by elution with 4M MgCl2. The anti-C1q immunosorbent only bound 14% and 12% of the same IC and only 64% were recovered by eluting with 4M MgCl2. The IC extracted by the anti-C3g immunosorbent included those capable of extraction by the anti-C1q. The anti-C3g monoclonal recognizes not only the iC3b fragment of C3 and will therefore bind IC with affinity for bovine conglutinin but also subsequent degradation products containing the C3g antigen. Its wide range of reactivity for IC plus its excellent recovery properties make it the immunosorbent of choice for isolating complement-fixing IC.