The antigenic specificity of different states of aggregation of tobacco mosaic virus protein

Monoclonal antibodies against a plant virus

“…I t was observed by MILTON and vA~¢ ~EGENMORTEL (13) that in highly purified TMV preparations, monomers were released spontaneously from the nucleocapsid so that in the preparation used for immunization as well as by probable degradation of the nucleocapsids in the immunized mouse also eapsid monomers can function as antigen. If one assumes the correctness of the multideterminant model suggested for TMV (13,15) antigenic determinants m a y be exposed by the capsid monomer which are hidden in polymers and in the assembled nucleoeapsid.…”

“…However, in monoclonal hybridoma mass cultures an incubation period of 1 hour with substrate was already sufficient for the detection of antibodies (Table 3). To ascertain that only TMV capsid monomers were employed in the respective ELISA the concentration of TMV protein was adjusted to 50 ~g/ml and the coating buffer to pH 9.6 (7,15).…”

Serological Methods in the Identification and Characterization of Viruses

“…If artifacts arising from temperature variations or unsuspected contaminants can be excluded, the formation of double bands by apparently single systems has usually been attributed to the fact that the antigen was degraded into fragments with different serological specificities. It is unlikely, however, that this explanation applies to the two bands commonly observed with the protein subunit of tobacco mosaic virus (Kleczkowski, 1961;Van Regenmortel and Lelarge, 1973). When this subunit is isolated in the monomeric state from a single band in SDS polyacrylamide gels, it still gives rise to double precipitin lines.…”

“…A possible explanation for this phenomenon is that antibody binding can induce, in the monomer, a conformational change similar to that observed during the polymerization of the subunit into capsids. Antibodies specific for each conformational state are known to be present in most antisera (Van Regenmortel and Lelarge, 1973), and it is possible that they induce mutually exclusive surface determinants in the monomer.…”

Examination of a New Cross-Linker in Monoclonal Antibody Reaction Against the C-Terminus of Tobacco Mosaic Virus Coat Protein

“…The coat protein was isolated according to the method described by Fraenkel-Conrat [11] and the monomers [12] used in ELISA.…”