Metabolic radiolabelling of Mycoplasma gallisepticum on Vero cells and radioimmunoprecipitation assay

A monoclonal antibody (MAb) A2 was produced against a major polypeptide of Mycoplasma gallisepticum with a molecular mass of 64 kDa. MAb A2 reacted in immunoblot at a titre of 10 4.33 and had a titre of 10 4.5 in an enzyme-linked immunosorbent assay. In a radioimmunoprecipitation assay (RIPA) using metabolic [ 35S]methionine radiolabelling of M. gallisepticum suspension in Vero cell culture, MAb A2 was able to precipitate the 64 kDa protein and another protein of 47 kDa. The present study involving [ 35S]methionine labelling of M. gallisepticum in Vero cells represents a novel approach for labelling and characterizing the conformation-dependent mycoplasmal antigens in a RIPA system.