Production of monoclonal antibodies to Shigella. Enzyme-linked immunosorbent assay for screening hybridoma antibodies with intact bacteria

A simple and rapid enzyme-linked immunosorbent assay (ELISA)-type assay has been developed to screen hybridoma supernatant fluids with whole viable or killed bacteria as the antigen. The optimum concentration of acetone-killed and dried cell antigen for coating was 25–100 μg/ml. Screening of hybridoma supernatant fluids against whole cells, both with and without fixation, was assessed and both were equally sensitive. The data indicate that bacteria] fixation is detrimental in ELISA probably because of loss of antigenic structure. A highly specific monoclonal antibody (laM3) was produced against Shigella flexneri la and was employed to optimize the assay procedure.