Monoclonal-based enzyme-linked immunosorbent assay and immunochromatographic rapid assay for dihydrostreptomycin in milk

Abstract

Monoclonal antibody (MAb) against streptomycin was prepared by using a streptomycin–keyhole limpet hemocyanin conjugate for the immunization of mice. Splenocytes from BALB/c immunized mice were fused with P3X63Ag8U.1 myeloma cells. This resulted in one hybridoma cell line. Using this MAb, we developed quantitative assays for dihydrostreptomycin by means of an enzyme-linked immunosorbent assay (ELISA). Fifty percent inhibition concentration (IC 50) for the MAb was 2.0 ng ml −1. The detection limit was 0.1 ng ml −1 and the standard deviations were 0.5–4.7% for intra-assay and 0.9–5.9% for inter-assay, respectively. The detection limit using peroxidase was 10 ng ml −1 in milk. Using the MAb produced, a rapid test kit based on the immunochromatographic method was developed. The detection limit using the kit was 100 ng ml −1 in milk.