CHARACTERIZATION OF POLYCLONAL AND MONOCLONAL-ANTIBODIES TO PHYSALIS MOTTLE (TYMO) VIRUS

Polyclonal antibodies were raised against the Physalis mottle virus (P hMV) and its denatured coat protein (PhMV-P). Analysis of the reactivi ty of the polyclonal antibodies with tryptic peptides of PhMV-P in dot -blot assays revealed that many of the epitopes were common to intact virus and denatured coat protein. Five monoclonal antibodies to the in tact virus were obtained using hybridoma technology. These monoclonal antibodies reacted well with the denatured coat protein. Epitope analy sis suggested that probably these monoclonal antibodies recognize over lapping epitopes. This was substantiated by epitope mapping using the CNBr digest of PhMV-P in western blots. All the five monoclonals recog nized the N-terminal 15 K fragment. Attempts to further delineate the specific region recognized by the monoclonals by various enzymatic cle avages resulted in the loss of reactivity in all the cases. The result s indicate that these monoclonals probably recognize epitopes within t he N-terminal 15 K fragment of the coat protein.