A novel solid-phase test to study the binding of IFN-gamma to its receptor.
A novel solid phase assay for interferon-gamma (IFN-gamma) binding to the human IFN-gamma receptor was developed. The receptor binding assay is carried out using a soluble form of the recombinant IFN-gamma receptor protein corresponding to the extracellular portion of the IFN-gamma receptor. Using different IFN molecules and anti-IFN monoclonal antibodies, we show that the specificity of the soluble IFN-gamma receptor coated to the plastic surface is not altered. In consequence, this new generation binding test can be used to characterize the interactions with the specific ligand under controlled conditions. In comparison with ELISA or RIA tests using antibodies specific for IFN-gamma, the solid-phase binding assay has the advantage of detecting only the active molecules. Finally, since the test has a large capacity, it is being applied for the screening of agents that are able to neutralize the IFN-gamma activity either by blocking the active site of the lymphokine or the binding site of the specific receptor.
Ozmen
L
L
P.R.T. Biology, Hoffmann-La Roche Ltd., Basel, Switzerland.
Fountoulakis
M
M
Garotta
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007158:Immunologic Techniques; D007371:Interferon-gamma / Q000378:metabolism*; D007372:Interferons / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D011863:Radioimmunoassay; D011971:Receptors, Immunologic / Q000378:metabolism*; D017471:Receptors, Interferon; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D008024:Ligands; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D007371:Interferon-gamma; D007372:Interferons