ABSTRACT

The ratio of hapten to bovine serum albumin (BSA) in an antigen conjugate was determined by matrix-assisted laser desorption/ionization-tof (MALDI-tof) mass spectrometry. A hybridoma secreting monoclonal antibody (MAb) was produced by fusing splenocytes immunized with an antigen-BSA conjugate with HAT-sensitive mouse myeloma cells. MAbs were screening and cloning, and used for ELISA. New Western blotting method of determination for solasodine glycosides and ginsenosides 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 glycoside-BSA conjugate on the PVDF membrane that was stained by the general Western blotting. Immunoaffinity column chromatography using anti-forskolin MAb appears to be far superior to previously published separation methods resulting in pure forskolin from the crude extracts of Coleus forskohlii root.