Monoclonal antibodies and enzyme immunoassays specific for human interferon (IFN) omega 1: evidence that IFN-omega 1 is a component of human leukocyte IFN.

Four hybridoma cell lines secreting monoclonal IgG antibodies to human interferon (IFN) omega 1 (= IFN-alpha II 1) were developed, using spleen cells of mice immunized with IFN-omega 1 and/or a novel hybrid interferon, IFN-omega 1/alpha 2. All antibodies (OMG-2, -4, -5, and -7) neutralize the antiviral activity of IFN-omega 1 and show distinct patterns of reactivity with the hybrid proteins, IFN-omega 1/alpha 2 and IFN-alpha 2c/omega 1. However, none of the antibodies is able to neutralize human IFN-alpha, confirming earlier observations that IFN-omega 1 and IFN-alpha are antigenically unrelated. The epitope specificities of the antibodies were further characterized in direct and competitive enzyme immunoassays (ELISAs). All binary antibody combinations were tested for their suitability for a two-site ("sandwich") ELISA for IFN-omega 1, using horse radish peroxidase as the marker enzyme. A configuration employing OMG-2 for antigen capture and OMG-7 as the detector antibody resulted in the highest assay sensitivity (approximately 10 pg IFN-omega 1/ml) and was studied further. This one-step assay is highly specific for IFN-omega 1 and does not recognize human IFN-alpha, -beta, and -gamma, thus allowing for determination of IFN-omega 1 levels in natural mixtures of human IFNs. Using this ELISA, it was found that IFN-omega 1 is present in IFN preparations derived from virus-induced human peripheral blood leukocytes and may constitute as much as 15% of the total leukocyte IFN activity. IFN-omega 1 was also detected at somewhat lower levels in preparations of human "lymphoblastoid" IFN.