Reisolation of immunoreactive radioiodinated antigens using glutaraldehyde-insolubilized antibody preparations.
Radioiodination of antigens for use in radioimmunoassay can result in substantial losses of antigenic reactivity with the corresponding antibody and antisera preparations. We describe a method whereby antigens iodinated with the chloramine-T procedure are bound to and eluted from glutaraldehyde-insolubilized antibody. Unfractionated antisera, an ammonium sulfate precipitated fraction or the IgG fraction of antisera may be used as insolubilized immunoadsorbents. The method has been applied for the reisolation of a radioiodinated peptide, a low molecular weight protein and the fibronectin molecule. The total binding of 125I-antigens to antibody in radioimmunoassays can be increased from such low amounts before reisolation that the assay is not feasible, to 85% above background binding, after adsorption and elution from the insolubilized antibody preparations.
D000327:Adsorption; D000447:Aldehydes / Q000276:immunology*; D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis; D000941:Antigens / Q000276:immunology*; D005976:Glutaral / Q000276:immunology*; D007457:Iodine Radioisotopes / Q000276:immunology*; D011817:Rabbits; D011863:Radioimmunoassay
D000447:Aldehydes; D000936:Antigen-Antibody Complex; D000941:Antigens; D007457:Iodine Radioisotopes; D005976:Glutaral