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. 1. Introduction Immunoassay systems using monoclonal antibody (MAb) against drugs and small molecular weight bioactive compounds have become an important tool for studies on receptor binding analysis, enzyme assay, and quantitative and/or qualitative analytical techniques in animals or plants. The immunoblotting method is based on the western blotting technique that utilizes the antigen-antibody binding properties and provides a specific and sensitive detection of higher molecule analytes like peptides and proteins. In our ongoing study on MAb, previously we prepared various kinds of MAb against natural products like forskolin [1], solamargine [2], crocin [3], marihuana compounds [4], opium alkaloids [5], ginsenosides [6, 7], berberine [8], sennosides [9], paeoniflorin [10], glycyrrhizin [11, 12], ginkgolic acid [13], aconitine alkaloid [14], baicalin [15], and so on and developed individual competitive enzyme-linked immunosorbent assay (ELISA) as a highly sensitive, specific, and simple methodology. The confirmation of hapten number in synthesized antigens is most important in the first stage of MAb preparation. Therefore, its determination method will be discussed first of all. As an application of MAb, the MAb against ginsenosides and glycyrrhizin has been prepared resulting in the development of two new techniques that we have named the eastern blotting method [12] and the knockout extract preparation [16]. They will be introduced in this paper. 2. Preparation of MAb against Natural
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