An improved technique for the isolation and analysis of immune complexes.
A technique for isolating and analyzing immune complexes (IC) using rheumatoid factor (RF) as immunoadsorbent is described. It contains several modifications to a previously published original method and has given greatly improved results: a small amount of BSA-anti-BSA (aBSA) model IC was added to human serum and the mixture was filtered through a column of Sephacryl S-300, the large molecular weight fraction was concentrated and added to tubes coated with RF. The bound IC were eluted, radioiodinated and freed to the major contaminants (HSA and immunoglobulins) by adding the corresponding antisera and removing the resulting complexes by coprecipitation or with the aid of protein A-containing Staphylococci. The purified preparations were analyzed by SDS-polyacrylamide gel electrophoresis (PAGE) and autoradiography. BSA was clearly identified in the autoradiograms. An example is given of the application of this technique to the isolation and analysis of IC from an abdominal effusion obtained from a patient with ovarian cancer.
D000005:Abdomen; D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000302:isolation & purification*; D001665:Binding Sites; D002417:Cattle; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D006333:Heart Failure / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007074:Immunoglobulin G; D010051:Ovarian Neoplasms / Q000276:immunology; D012217:Rheumatoid Factor / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000276:immunology; D013997:Time Factors
D000936:Antigen-Antibody Complex; D007106:Immune Sera; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D012217:Rheumatoid Factor