A two-site sandwich radioimmunoassay of human gamma interferon with monoclonal antibodies.

Two monoclonal antibodies (nos. 6008 and 6016) were raised against human gamma interferon (IFN-gamma) derived from E. coli harboring the recombinant cDNA for IFN-gamma, and one (3710) against a synthetic peptide representing its C-terminus amino acid sequence of 20 residues. The monoclonal antibody against the synthetic peptide (3710) reacted either with IFN-gamma or the synthetic peptide. One monoclonal anti-IFN-gamma (6008) did not react with the synthetic peptide, while the other (6016) showed a weak binding with the peptide. The binding of the monoclonal antibody against the synthetic peptide (3710) with IFN-gamma was not inhibited by 6008, but to a certain extent by 6016. A 2-site '1-step' radioimmunoassay was developed in which 6008 was fixed on a solid-phase support, and the test sample together with radiolabeled 3710 was added for the binding with it. The assay was rapid with a sensitivity capable of detecting a few ng/ml of IFN-gamma.