Influence of immune-complex size and antigen-antibody ratio on immune complex detection with monoclonal rheumatoid factor and C1q.
Stabilized aggregates of human IgG were prepared over a wide range of molecular weights. These fractions with increasing molecular weight were adjusted to the same molarity or the same protein concentration and were tested in the solid phase C1q and monoclonal rheumatoid factor immune complex assays. At constant molarity results were linearly correlated with the size of the aggregates. At constant protein concentration, results were also linearly correlated with the size of the aggregates in the lower molecular weight fractions, although the number of aggregates in the fractions decreased with increasing molecular weight. It is concluded that results in these assays can only be compared with respect to concentration if the immune complexes have identical sizes. Consequently, we studied the relative affinity of C1q and monoclonal rheumatoid factor for antigen/antibody immune complexes of different sizes or different antigen/antibody ratios, using model immune complexes composed of tetanus toxoid/anti-toxoid and streptolysine O/anti-streptolysine O. Compared to C1q, monoclonal rheumatoid factor was found to have higher affinity for smaller complexes, and for complexes with higher antigen/antibody ratio. Immune complex containing sera of 5 patients with connective tissue diseases, and one normal serum, were fractionated on a Sepharose 4B column. The binding patterns of the different fractions to solid phase C1q and monoclonal rheumatoid factor were quite variable and in only one of these sera monoclonal rheumatoid factor had the expected preference for small complexes and C1q for large complexes. Moreover a remarkably high binding to C1q and/or monoclonal rheumatoid factor of the monomeric IgG fraction was found in 4 out of 5 patient sera.
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis*; D000937:Antigen-Antibody Reactions; D003166:Complement Activating Enzymes; D015922:Complement C1q; D003240:Connective Tissue Diseases / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G; D008970:Molecular Weight; D012217:Rheumatoid Factor
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D015922:Complement C1q; D012217:Rheumatoid Factor; D003166:Complement Activating Enzymes