Development of an enzyme-linked immunosorbent assay (ELISA) using highly-specific monoclonal antibodies against plumbagin |
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Authors: | Sakamoto Seiichi Putalun Waraporn Tsuchihashi Ryota Morimoto Satoshi Kinjo Junei Tanaka Hiroyuki |
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Affiliation: | a Department of Medicinal Plant Breeding, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan b Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 4002, Thailand c Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka 814-0180, Japan |
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Abstract: | Plumbagin (PL; 5-hydroxy-2-methyl-1,4-naphthoquinone) is a natural compound mainly isolated from Plumbago zeylanica. This plant is distributed in Southeast Asia, and well known as Ayurvedic medicine in India for its medicinal properties. PL has been shown to have various pharmacological activities. We have successfully prepared monoclonal antibodies against PL, and developed an enzyme-linked immunosorbent assay (ELISA) system for determination of PL. 3-(5-Hydroxy-2-methyl-1,4-naphthoquinone-3-yl) propanoic acid was synthesized and purified to prepare PL-bovine serum albumin conjugate (PL-BSA), which was used as an immunogen. PL-BSA conjugate was administered into BALB/c male mice for production of monoclonal antibodies against PL. The monoclonal antibody against PL which is secreted from established hybridoma cell line 3A3 (MAb 3A3) has been proven to have highly-specific to PL resulting from cross-reactivities test. The range for calibration of PL by ELISA was 0.2-25 μg mL−1. Based on validation analysis, this analytical method by ELISA is a precise, accurate, and sensitive method for the determination of PL in plant. |
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Keywords: | Plumbagin Plumbago zeylanica Enzyme-linked immunosorbent assay Monoclonal antibody |
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