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Characterization of Murine Monoclonal Antibodies to Human Interferon‐Gamma (IFN‐ γ ) and Their Application for Sandwich Enzyme‐Linked Immunosorbent Assay (ELISA)
Author(s) -
Jitsukawa Tomofumi,
Nakajima Satoko,
Sugawara Isamu,
Mori Shigeo,
De Ley Marc
Publication year - 1987
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.1987.tb03142.x
Subject(s) - monoclonal antibody , interferon gamma , antibody , microbiology and biotechnology , epitope , recombinant dna , biology , interferon , virology , cytokine , immunology , biochemistry , gene
The three murine monoclonal antibodies (MAb), D1G2, D9D10, and D13C8, are specific for human interferon‐ γ (IFN‐ γ ), but not human IFN‐ α and IFN‐ β . They react weakly with heat‐treated IFN‐ γ . The three antibodies recognize different epitopes of the IFN‐ γ 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‐ γ 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‐ γ exists in oligomeric form. Recombinant human IFN‐ γ expressed in E. 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‐ γ molecule as well as measurement of IFN‐ γ in human sera and tissue culture fluids.