Production of antiserum to respiratory syncytial virus polypeptides: application in enzyme-linked immunosorbent assay

By use of crossed immunoelectrophoresis techniques, respiratory syncytial (RS) virus-specific precipitates were produced between RS virus cellular antigen [solubilized in tris(hydroxymethyl)aminomethane-glycine buffer, pH 9] and antiserum raised in rabbits against semipurified RS virus. When these precipitates were employed as antigens for further immunizations in rabbits, antibodies (anti-RSV-precip.I) were produced which reacted with only one RS virus antigen when tested by the crossed immunoelectrophoresis technique. Precipitates obtained between RS virus cellular antigen (labeled with L-[35S]methionine) and anti-RSV-precip.I were examined by polyacrylamide gel electrophoresis, which showed that anti-RSV-precip.I precipitated RS virus polypeptides of molecular weights 28,000 to 84,000. Anti-RSV-precip.I was employed as capture antibodies in the enzyme-linked immunosorbent assay, in which RS virus cellular antigen was used as the second layer. Determination of human RS virus immunoglobulin G antibodies by this enzyme-linked immunosorbent assay technique showed a high degree of sensitivity, specificity, and reproducibility.