A Radioimmunoassay for Virus Antibody using Binding of 125I-labelled Protein A

Abstract

An assay for virus antibodies using protein A from Staphylococcus aureus is described. Type B and type C RNA tumour viruses adsorbed on to polystryrene microtitre plate wells were incubated with antiserum and then with ¹²⁵I-labelled protein A (I-pA) and bound radioactivity was determined. Technical details such as labelling, antigen concentration, storage of I-pA are reported. The specificity of the reaction was investigated in detail by competition experiments with purified unbound homologous viruses. This assay also proved to be sensitive for demonstration of autogenous immunity to both type B and type C RNA tumour viruses. A study using antisera against purified core and envelope virus proteins of mammary tumour and leukaemia viruses suggested that the reaction mainly involves surface antigens of the intact virions.