Abstract
Hybridoma cells producing monoclonal antibodies were prepared by fusion of mouse myeloma cells and spleen cells derived from a BALB/c mouse immunized with purified preparations of a Japanese common strain of tobacco mosaic virus (TMV-OM). Ascites fluid containing high-titred antibodies against TMV-OM was obtained after injecting 18 cloned hybridoma lines into the peritoneal cavity of pristan-primed BALB/c mice. Antibody titres of these ascites fluid were 100 to 1, 000-fold higher than the titres of culture fluid derived from the hybridoma and had an end point of 1.6×107 in the passive hemagglutination test. Using these monoclonal antibodies, antigenicity of TMV-OM was analyzed by double-immunodiffusion, reverse passive hemagglutination inhibition, and passive hemagglutination. Of 18 monoclonal antibodies from hybridoma lines against TMV-OM, 5 were specific for TMV-OM, 9 reacted with both TMV-OM and a tomato strain of TMV (TMV-T), and 4 were specific for TMV-T. These results indicated that at least three antigenic determinants are present in TMV-OM. In the further test for the monospecificity of these three monoclonal antibodies against. TMV-OM using passive hemagglutination inhibition, it was found that of 6 epitopes, two are associated with TMV-OM specific antibodies, three for antibodies specific to both TMV-OM and TMV-T, and one for TMV-T specific antibodies.
