Immunoquantitative analysis for berberine and its related compounds using monoclonal antibodies in herbal medicines.

The monoclonal antibody (MAb) against berberine, a bioactive constituent of Coptis japonica M., Phellodendron amurense R. and Hydrastis canadensis L., was produced and characterized. As immunogen, the derivative of berberine, 9-O-carboxymethyl berberrubine was synthesized and conjugated to carrier protein, bovine serum albumin (BSA). In order to confirm its immunogenicity, the ratio of hapten in berberine-BSA conjugate was determined by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). After immunization, hybridomas secreting MAbs against berberine were produced by fusing splenocytes with mouse myeloma cell line, P3-X63-Ag8-653. After the screening, anti-berberine MAb 1D5-3B-7 was obtained. Subsequently, a quantitative ELISA system for berberine and its related compounds using the MAb was established and evaluated comparing with HPLC method. The ELISA method described in this study can be available as an analytical tool for quality control and standardization of medicinal plants and its prescriptions containing berberine and its related compounds.