title: Application of Monoclonal Antibodies against Bioactive Natural Products: Eastern Blotting and Preparation of Knockout Extract.
abstract: Matrix-assisted laser desorption/ionization (MALDI) tof mass spectrometry was used for the confirmation of hapten number in synthesized antigen. As application of MAb, the MAbs against ginsenosides and glycyrrhizin have been prepared resulting in the development of two new techniques that we named the eastern blotting method and the knockout extract preparation. In eastern blotting technique, glycosides like ginsenosides and glycyrrhizin separated by silica gel TLC were blotted to PVDF membrane that was treated with a NaIO(4) solution followed by BSA resulted in glycoside-BSA conjugate on a PVDF membrane. The blotted spots were stained by MAb. Double staining of eastern blotting for ginsenosides using antiginsenoside Rb(1) and Rg(1) MAbs promoted complete identification of ginsenosides in Panax species. The immunoaffinity concentration of glycyrrhizin was determined by immunoaffinity column conjugated with antiglycyrrhizin MAb resulting in the glycyrrhizin-knockout extract, which was determined by the synergic effect with glycyrrhizin on NO production using the cell line.
authors.affiliation: Graduate School of Pharmaceutical Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-0855, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Osamu
authors.initials: O
authors.lastname: Morinaga
authors.forename: Takuhiro
authors.initials: T
authors.lastname: Uto
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Fuji
authors.forename: Frederick Asare
authors.initials: FA
authors.lastname: Aboagye
authors.forename: Nguyen Huu
authors.initials: NH
authors.lastname: Tung
authors.forename: Xiao Wei
authors.initials: XW
authors.lastname: Li
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Shoyama