The Detection of Antigens in Immune Complexes
- 1 May 1981
- journal article
- research article
- Published by Wiley in Scandinavian Journal of Immunology
- Vol. 13 (5), 441-446
- https://doi.org/10.1111/j.1365-3083.1981.tb00154.x
Abstract
A procedure is described for the purification of soluble immune complexes (IC) from biological fluids and for the detection of antibody-bound antigen. A model IC, prepared with various amounts of human serum albumin (HSA) and constant amounts of anti-HSA, gave a 44% recovery after gel filtration on Sephacryl S-300, affinity chromatography on protein A-Sepharose, and concentration of eluates therefrom before analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was calculated that 20 ng of antibody-bound HSA should be detectable when polyacrylamide gels are subjected to a direct radioimmunoassay involving the use of rabbit anti-HSA, 125I-protein A and autoradiography, thus defining the sensitivity of the procedure for detecting IC-bound antigen in a model system. The procedure has direct relevance to the examination of IC of unknown composition present in sera of individuals with various diseases.This publication has 18 references indexed in Scilit:
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