The major internal protein of the RD-114 virus that appeared in a human tumor cell line, RD, after passage through fetal cats was purified by isoelectric focusing and used to prepare antibody in guinea pigs.

The major internal protein of a rat type C virus pseudotype of murine sarcoma virus, MSV(RaLV), was purified by isoelectric focusing (pl = 8.6) and used to prepare antibody in guinea pigs. The protein was identified by its reaction with antisera reactive with the mammalian type C virus group-specific (gs) antigenic determinant, gs-3. The guinea pig antisera mainly contained species-specific (gs-1) antibody for reactions in gel diffusion with other type C viruses were limited to those of rat origin, whereas in complement fixation tests heterologous reactions could be eliminated by use of appropriate antiserum concentrations without affecting homologous reactions. Guinea pig antisera against mouse, hamster, or cat gs-1 determinants did not react with MSV(RaLV) purified gs protein or with any of several other rat type C viruses.

The mammalian C-type tumor viruses share an antigenic determinant, gs-3, located on the major internal polypeptide of the virion. Detection of this determined in gel diffusion assays by antiserums prepared in rats by immunization with rat tumor homogenates carrying murine virus and serums prepared in a rabbit by immunization with purified murine gs antigen depended on antibodies present in the fractions containing immunoglobulins M and G. The immunoglobulin G fraction by itself precipitated only the homologous murine antigen. Neither fraction alone precipitated heterologous (cat, rat, or hamster) antigen (definition of the gs-3 reaction), while a mixture of the two fractions did. The gs-3 reaction was eliminated by treatment of the serums with beta-mercaptoethanol, also indicating a requirement for immunoglobulin M antibodies.