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The Detection of Antigens in Immune Complexes
Author(s) -
HEIMER R.,
GLICK D. L.,
ABRUZZO J. L.
Publication year - 1981
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1981.tb00154.x
Subject(s) - chromatography , radioimmunoassay , antigen , size exclusion chromatography , antibody , chemistry , sepharose , polyacrylamide gel electrophoresis , human serum albumin , immunoprecipitation , immune complex , affinity chromatography , bovine serum albumin , monoclonal antibody , immune system , gel electrophoresis , biochemistry , biology , immunology , enzyme
A procedure is described for the purification of soluble immune complexes (IC) from biologica 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 sulphate‐polyacryl‐amide gel electrophoresis. It was calculated that 20 ng of antibody‐bound HSA should be detectable when polyacrylamide gets are subjected to a direct radioimmunoassay involving of the use of rabbit anti‐HSA. 125 I‐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.