Changes in antibody avidity after virus infections: detection by an immunosorbent assay in which a mild protein-denaturing agent is employed

Abstract

In titrating serum immunoglobulin G antibody to viruses by enzyme-linked immunosorbent assay, we used two rows of wells for serial twofold dilutions of the serum; in one row, a low concentration of a protein denaturant, 0.5 or 1.0 M guanidine hydrochloride, was added to the diluent so that the binding of low-avidity antibodies to viral antigens on the solid phase was inhibited. We then compared the antibody titration curves obtained in the two rows. We found that the addition of the reagent resulted in a parallel leftward shift of the curves and that the extent of the shift was greater in early than in late sera from all of the three infections studied (Japanese encephalitis virus, rotavirus, and rubella virus infections). This procedure may be useful for estimation of the avidity of antibody in serum and, with further evaluation, may prove to be applicable to single-serum diagnosis of virus infections.