Characterization of Murine Monoclonal Antibodies to Human Interferon‐Gamma (IFN‐γ) and Their Application for Sandwich Enzyme‐Linked Immunosorbent Assay (ELISA)

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Abstract
The three murine monoclonal antibodies (MAb), D1G2, D9D10, and D13C8, are specific for human interferon-.gamma. (IFN-.gamma.), but not human IFN-.alpha. and IFN-.beta.. They react weakly with heat-treated IFN-.gamma.. The three antibodies recognize different epitopes of the IFN-.gamma. molecule, as evaluated by antibody-binding inhibition experiments. We have used these three monoclonal antibodies to construct a sandwich enzyme-linked immunosorbent assay (ELISA). The best result was obtained when we used D1G2 or D9D10 MAb as a solid-phase immunosorbent and D1G2 or D9D10 MAb as a tracer. When we measured IFN-.gamma. in sera by a combination of D1G2 (a solid-phase) and D1G2 (a tracer), a result similar to the one by a combination of D9D10 (a solid-phase) and D1G2 (a tracer), was obtained. This may suggest that human IFN-.gamma. exists in oligomeric form. Recombinant human IFN-.gamma. expressed in Escherichia coli is detectable at a concentration of 1 ng/ml in this sandwich ELISA. This assay can be employed for the analysis of the structural characteristics of the human IFN-.gamma. molecule as well as measurement of IFN-.gamma. in human sera and tissue culture fluids.