Detection and differentiation of immune complexes and IgG aggregates by a complement consumption assay.
The applicability of a complement consumption assay as a means by which to detect IgG aggregates and immune complexes in serum was examined. Both heavy (greater than or equal to 19S) and intermediate (11-17S) IgG aggregates were detected and the sensitivity of the assay was greater than or equal to 10 mug aggregated IgG/ml. BSA anti-BSA complexes, formed in slight antibody excess, were detected at a BSA concentration of 200 ng/ml. NHS stored at 4degreesC for greater than or equal to 2-3 weeks or at -20degreesC for more than 3 months developed distinct anticomplementarity (AC). This background AC, due to IgG aggregate formation, was reduced by heating the serum at 56degreesC for 50 min prior to testing. A similar reduction of AC and C1q fixation was observed when IgG aggregated at 61degreesC or 63degreesC was heated further at 56degreesC for 50 min. The abatement of AC could not be correlated to a change in IgG aggregation size. In contrast, AC of preformed antigen-antibody complexes was not reduced by this heat treatment.
D000936:Antigen-Antibody Complex; D002850:Chromatography, Gel; D003165:Complement System Proteins; D006358:Hot Temperature; D006801:Humans; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000032:analysis; D012710:Serum Albumin, Bovine / Q000276:immunology*; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D003165:Complement System Proteins