A comparison of ELISA screening methods for the production of monoclonal antibodies against soluble protein antigens.

A library of monoclonal antibodies against pig heart citrate synthase has been raised. Twelve solid-phase immunoassay systems, employing different methods of antigen immobilization, have been compared for their ability to detect the various members of this library. It was found that a sandwich immunoassay, in which the citrate synthase antigen was immobilized on the solid support via a polyclonal antibody preparation, was the only system capable of detecting all the monoclonal antibodies tested. It is suggested that such a sandwich assay should be used in preference to direct assays for the initial screening of monoclonal antibodies.