Monoclonal capture antibody ELISA for respiratory syncytial virus: detection of individual viral antigens and determination of monoclonal antibody specificities

Abstract

An enzyme-linked immunosorbent assay (ELISA) for respiratory syncytial virus (RSV) that employs solid-phase monoclonal antibodies was developed. RSV antigens bound by these monoclonal capture antibodies were detected by addition of a polyclonal bovine antiserum, followed anti-bovine enzyme conjugate and enzyme substrate. The sensitivity and specific of the assay were determined by titrations of the solid-phase antibodies and by antigen titrations with both unpurified RSV-infected cell culture material and purified RSV nucleocapsids. The addition of a competitive binding step prior to the addition of antigen to the solid-phase antibody provides further evidence of the assay's specificity. Furthermore, the competitive binding assay enables the antigen specificity of monoclonal antibodies to be determined or compared without the use of purified antigens. Monoclonal capture ELISA is a convenient, rapid, and sensitive assay that can be used to measure specific RSV antigens in unpurified preparations as well as to determine anti-RSV antibody specificity and should prove useful in examining other complex antigen-antibody systems.