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International Journal of Analytical Chemistry
Volume 2012 (2012), Article ID 260425, 10 pages
http://dx.doi.org/10.1155/2012/260425
Review Article

Application of Monoclonal Antibodies against Bioactive Natural Products: Eastern Blotting and Preparation of Knockout Extract

1Graduate School of Pharmaceutical Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-0855, Japan
2Faculty of Pharmaceutical Science, Nagasaki International University, 2827-7 Huis Ten Bosch, Sasebo 859-3298, Japan
3Faculty of Health Management, Nagasaki International University, 2827-7 Huis Ten Bosch, Sasebo 859-3298, Japan
4Phytochemistry Department, Centre for Scientific Research into Plant Medicine, University of Ghana, P.O. Box 73, Mampong-Akuapem, Ghana
5State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No.38 Xue-yuan Road, Haidian District, Beijing 100191, China
6Faculty of Pharmaceutical Science, Khon Kaen University, Khon Kaen 40002, Thailand

Received 30 August 2011; Accepted 18 October 2011

Academic Editor: Norberto Peporine Lopes

Copyright © 2012 Hiroyuki Tanaka et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 NaIO4 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 Rb1 and Rg1 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.