Detection of monoclonal influenza antibodies synthesized in culture by hybridoma cells with a solid-phase indirect immunofluorometric assay

Abstract

A solid-phase indirect immunofluorometric assay for measuring reactions of mouse monoclonal antibodies with antigen has been developed, with influenza virus as a model. Purified IgG from hyperimmune rabbit sera is covalently linked to polyaminostyrene beads, to which influenza viruses are then bound immunologically to make solid-phase antigens. Alternatively, the virus is covalently coupled directly to the beads. Mouse antibodies, produced by hybridoma cells in culture, are reacted with constant amounts of solid-phase antigens, and then indirectly quantitated by adding FITC-labeled antimouse Ig and measuring the fluorescent intensity with a filter-fluorometer. The assay system permits rapid screening for low levels of antibodies synthesized by hybridoma cells in culture. It is about 25- to 150-fold more sensitive than hemagglutination inhibition tests in detecting monoclonal antibodies reactive with influenza virion HA protein.