Radioimmunoassay evaluation of anti‐C3d reactivity in broad spectrum commercial antiglobulin reagents

Abstract

The study was undertaken to assess the potential of a labeled C3d radioimmunoassay method as a standardization criterion for antiglobulin reagents containing anti-C3d reactivity. Over an 18-month interval, four different lots of broad-spectrum antiglobulin reagents were purchased from each of seven United States manufacturers. Anti-C3d antibody concentration and Ko in each lot were assayed initially and at four-month intervals over 16 to 21 months storage at 4 degrees C. The results permitted 1) comparison of anti-C3d antibody concentrations among different manufacturers, 2) comparison among different lots from the same manufacturer, and 3) assessment of stability of anti-C3d during storage. In addition, serial dilutions of reagents in each manufacturer's diluent were compared for their anti-C3d agglutinating properties by the spin-antiglobulin method against red blood cells (RBC) coated by C3d in vitro and against in vivo C3d-coated RBC from 16 patients. Most reagents were shown also to contain anti-C3c, anti-C4c and anti-C4d reactivities when tested against suitably coated RBC. Anti-C3d antibody concentrations ranged from approximately 1 to 3.5 microgram/ml for 27 of the 28 reagents and were stable over 16 to 21 months of storage. Anti-C3d agglutination titer scores showed a general correlation with anti-C3d antibody concentrations when tested against in vitro C3d-coated RBC; correlations were less good against C3d-coated cells from patients. The radioimmunoassay provided reproducible objective measurements and appeared to have merit as a standardization criterion.