MAPPING OF THE DOMINANT NEUTRALIZING ANTIGENIC SITE OF A VIRUS USING INFECTED-CELLS

Panels of neutralizing monoclonal antibodies (MAbs) and antisera to ve sicular stomatitis virus of the serotype Indiana (VSV-IND) were genera ted in mice and rats. They were used in competition studies to map epi topes on the viral glycoprotein that are involved in virus neutralizat ion. Since neutralizing antibodies bind to the viral glycoproteins on the surface of intact viruses and of infected cells, infected cells we re used for measuring the binding of competing antibodies by cytofluor ometric analysis. A single immunodominant neutralizing epitope was rec ognised by 90% (58) of the MAbs including all of strong neutralizing c apacity. 10% (6) of the neutralizing MAbs that all exhibited low neutr alizing titers. recognised spatially closely related epitopes. This ap proach offers a convenient method to determine antibody interaction wi th complex conformational epitopes of membrane proteins.