Radioimmunoprecipitation of avian reovirus polypeptides using virus-specific IgM and IgG murine monoclonal and chicken polyclonal antibodies.
A simple and improved procedure for radioimmunoprecipitation (RIPA) for the identification of major immunogenic proteins of avian reovirus using murine monoclonal and chicken polyclonal antibodies is described. Bacterial proteins (Staphylococcus aureus-Protein A or Streptococcus species-Protein G) commonly used in RIPA procedures lack reactivities with low avidity monoclonals belonging to immunoglobulin (Ig)G or IgM subtypes, as well as avian Ig. Hence we used an indirect approach utilizing species-specific anti-IgM, IgG or chicken Ig antibodies followed by the precipitation of the immune complexes using Protein A. This improved the sensitivity of the RIPA enabling the antigenic analysis of the major antigenic proteins of avian reovirus. The results indicated that the virus is highly immunogenic in the natural host, chicken than in mice. Furthermore, there exists a direct correlation between a strong neutralizing antibody response and an increased precipitation of the sigma (sigma) proteins of the virus. The results also demonstrate a strong association between the conformational viral epitopes of the three classes of proteins, large (lambda), medium (mu) and small (sigma).
Reddy
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Département de pathologie et microbiologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Quebec, Canada.
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests / Q000379:methods; D011233:Precipitin Tests / Q000379:methods*; D011863:Radioimmunoassay / Q000379:methods*; D012087:Reoviridae; D014764:Viral Proteins / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D014764:Viral Proteins