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.
Silim
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Département de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, St-Hyacinthe, Quebec.
Kheyar
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D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology*; D002522:Chlorocebus aethiops; D007553:Isotope Labeling; D008715:Methionine; D009174:Mycoplasma / Q000276:immunology*; D015531:Radioimmunoprecipitation Assay / Q000379:methods*; D013462:Sulfur Radioisotopes; D014709:Vero Cells
D000942:Antigens, Bacterial; D013462:Sulfur Radioisotopes; D008715:Methionine