Pharmacologically Active Saikosaponin in Bupleurum falcatum Detected by Competitive ELISA and Eastern Blotting Using Monoclonal Antibodies

Abstract

This study showed the effective microanalytic methods for detection of saikosaponin by a competitive enzyme-linked immunosorbent assay (ELISA) using anti-saikosaponin a (SSa) monoclonal antibody (MAb). The results showed that the competitive ELISA had higher sensitivity than high performance liquid chromatography (HPLC) for the detection of saikosaponin. ELISA showed a highly significant difference in SSa concentrations between the roots of Bupleurum falcatum with different origins. In addition, a difference in SSa concentrations was observed between cytologically different cultivars originating from Japan (2n=26) and Korea (2n=20). It also showed excellent performance in quantitative analysis of SSa in the small samples from regenerated plantlets and calli. In addition, Eastern blotting using anti-SSa MAb 3G10 multispecific to saikosaponins was applied to analyze the distribution of saikosaponins in Bupleurum falcatum roots and leaves. The localized distribution of saikosaponins in whole leaves and cork layer of the roots in B. falcatum was successfully demonstrated by this novel immunoassay.