A protein A-binding, polyethylene glycol precipitation-based immunoradiometric assay. Application to the detection of immune complexes and C3 in human sera and of private antigens in cross-reacting parasite extracts.
An immunoradiometric assay, based on the precipitation of antigen-antibody complexes by polyethylene glycol (PEG) and on the subsequent binding of PEG-soluble radiolabelled staphylococcal protein A to the PEG-insoluble complexes, is described. The assay can be applied to the detection of naturally occurring, circulating immune complexes, and of complexes artificially created by mixing antigen and antibody solutions, which makes it of potential use for the detection of either antigen or antibody in several situations. Pre-treatment of the antibody-containing sera with 3% PEG greatly reduced the background values and increased the sensitivity of the assay. The assay was also applied to the detection and isolation of Leishmania donovani antigens that did not cross-react with antigens of the related parasite Trypanosoma cruzi (private antigens) and private antigens of insect-derived metacyclic trypomastigotes of T. cruzi in relation to culture-derived metacyclic trypomastigotes of T. cruzi. A simple and extremely effective procedure for washing precipitates with just one centrifugation is also described.
D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis*; D000953:Antigens, Protozoan / Q000276:immunology; D003176:Complement C3 / Q000032:analysis*; D006801:Humans; D007893:Leishmania donovani / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000276:immunology; D011092:Polyethylene Glycols; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D013205:Staphylococcal Protein A / Q000276:immunology*; D014349:Trypanosoma cruzi / Q000276:immunology
D000936:Antigen-Antibody Complex; D000953:Antigens, Protozoan; D003176:Complement C3; D013205:Staphylococcal Protein A; D011092:Polyethylene Glycols