A small scale indirect 125iodine-labelled protein A binding assay for detection of monoclonal antibodies against avian oncoviral proteins.
A small scale solid-phase radioimmunoassay employing 125I-labelled protein A is described which is suitable for screening large numbers of monoclonal antibodies directed against antigens which can be prepared in small amounts only, for example oncoviral proteins. The use of polystyrene Terasaki microtest plates instead of 96-well microtitre plates reduces the amount of antigen required for screening hybridoma supernatants to less than 30 ng/well. This method facilitates washing procedures and reduces the quantity of radioactive waste. The sensitivity and specificity of the method is demonstrated by the isolation and initial characterization of monoclonal antibodies specific for avian oncoviral transforming, structural and polymerase proteins.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000276:immunology; D001358:Avian Sarcoma Viruses / Q000276:immunology*; D007457:Iodine Radioisotopes; D051379:Mice; D008815:Mice, Inbred Strains; D015688:Oncogene Protein pp60(v-src); D011494:Protein Kinases / Q000276:immunology*; D012194:RNA-Directed DNA Polymerase / Q000276:immunology*; D013205:Staphylococcal Protein A; D014764:Viral Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D007457:Iodine Radioisotopes; D013205:Staphylococcal Protein A; D014764:Viral Proteins; D011494:Protein Kinases; D015688:Oncogene Protein pp60(v-src); D012194:RNA-Directed DNA Polymerase