ELISA, western blotting, immunocytolocalization and immunoaffinity column for naturally occuring bioactive compounds using monoclonal antibodies

Anti-sennoside A (SA) and anti-glycyrrhizin (GC) MAbs were used for screening and cloning. And they were also used for ELISA. New western blotting method of determination for GC and ginsenoside was established that compounds separated by silica gel TLC were transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was treated with NaIO₄ solution followed by BSA, resulting in a ginsenoside-BSA conjugate on the PVDF membrane which was stained by the general western blotting. Immunocytolocalization of GC in the fresh root of Glycyrrhiza species was conducted using anti-GC MAb after blotting to PVDF membrane. Immunoaffinity column chromatography using anti-ginsenoside Rb1 (GRb1)MAb performs better than previously published separation methods resulting in pure GRb1 from the crude extracts of ginseng. On the other hand, total solasodine glycosides were separated by an immunoaffinity column using a wide-cross reactive anti-solamargine (SM)MAb.