TY - JOUR
T1 - Development of an enzyme-linked immunosorbent assay (ELISA) using highly-specific monoclonal antibodies against plumbagin
AU - Sakamoto, Seiichi
AU - Putalun, Waraporn
AU - Tsuchihashi, Ryota
AU - Morimoto, Satoshi
AU - Kinjo, Junei
AU - Tanaka, Hiroyuki
N1 - Funding Information:
The research in this paper was supported in part by a Grant-in-Aid from JSPS's Asian CORE Program, the Ministry of Education, Culture, Sports, Science and technology of Japan, the research fund of Kyushu University Foundation and National Center for Genetic and Biotechnology (BIOTEC), Thailand.
PY - 2008/1/21
Y1 - 2008/1/21
N2 - 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.
AB - 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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U2 - 10.1016/j.aca.2007.11.021
DO - 10.1016/j.aca.2007.11.021
M3 - Article
C2 - 18155415
AN - SCOPUS:37249018589
SN - 0003-2670
VL - 607
SP - 100
EP - 105
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
IS - 1
ER -