Detection of immune complexes using a solid-phase C1q polystyrene ball assay.
A polystyrene ball C1q solid-phase assay (PSB C1q SPA) has been developed for quantitating immune complexes in human serum. While similar to the previously reported solid-phase C1q assays in principle, the use of polystyrene balls with a specular finish has resulted in an assay with significantly improved accuracy, sensitivity and reproducibility. The sensitivity of the assay based on the amount of AHGG bound per microgram C1q added was approximately 12-fold higher in the PSB assay compared to the polystyrene tube solid-phase C1q method. In order to correct for variable background contributions in different samples, values obtained with heat-inactivated C1q were subtracted from each experimental result. Reproducibility studies yielded a coefficient of variation (CV) of 10 and 4% in day-to-day assays using 25 microgram and 100 microgram AHGG/ml normal serum, respectively compared to 11--15% for the tube technique. Within run measurements gave a CV of 37 and 20% at the low and high levels of AHGG. Aggregated human gamma-globulin (AHGG) was used as a model immune complex and when chromatographed on Biogel A-15m yielded major fractions at greater than or equal to 15,000,000 and 150,000 daltons. Maximum binding of AHGG to PSB C1q occurred with aggregates greater than 15,000,000 daltons. Optimum binding of human albumin-anti-albumin complexes in the PSB C1q SPA occurred at a molar ratio of 1 : 1.5. The size distribution of this complex active in the assay determined by sucrose density gradients was 14--32 S with peaks at 21 and 27 S. A normal range of immune complexes was determined as 15 +/- 8 microgram AHGG equivalents (+/- 2 S.D., n = 65). Approximately 70% of rheumatoid arthritis, linear scleroderma, vasculitis, Sjögren's and glomerulonephritis and 40% of SLE patients were above 2 S.D. of normal. SLE patients demonstrated elevations in immune complexes only during periods of increased disease severity. The assay was a useful monitor of plasmapheresis in an SLE patient, showing decreases in immune complexes after each plasmapheresis. DNA did not interfere with AHGG binding whereas lipemia prevented detection of added AHGG.
D000936:Antigen-Antibody Complex; D001793:Blood Preservation; D001800:Blood Specimen Collection; D003172:Complement C1 / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods*
D000936:Antigen-Antibody Complex; D003172:Complement C1