Development of an enzyme-linked immunosorbent assay for the detection of isomiroestrol, an identical marker, in White Kwao Krua using a monoclonal antibody.
Pueraria candollei var. mirifica or White Kwao Krua (WKK) is a phytoestrogen-rich plant widely used among women to improve climacteric symptoms. Additionally, the tuberous roots of this plant have been added as an active ingredient for skin rejuvenation and breast enlargement effects in various functional foods. However, most of the products on the market containing WKK have not been sufficiently standardized with respect to the active compound or identical marker. To control the quality of these plant materials, an enzyme-linked immunosorbent assay (ELISA) using anti-isomiroestrol antibodies was established for the determination of isomiroestrol, an identical marker in WKK. Polyclonal and monoclonal antibodies against isomiroestrol were generated and their specificity characterized in this study. Monoclonal antibody 12C1 showed higher specificity to isomiroestrol and was thus selected to develop the ELISA. Based on the validation analysis and the tested performance of the developed ELISA in variably sourced WKK samples, the assay can provide an alternative approach that is reliable and highly sensitive for the quantitative analysis of isomiroestrol in plant.
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand.
Tharita
T
Kitisripanya
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand.
Kamonthip
K
Jutathis
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand.
Chadathorn
C
Inyai
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand.
Jukrapun
J
Komaikul
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand.
Orapin
O
Udomsin
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Hiroyuki
H
Tanaka
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand; Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University-Khon Kaen University, Khon Kaen, Thailand. Electronic address: waraporn@kku.ac.th.
Waraporn
W
Putalun
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015415:Biomarkers / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D048789:Phytoestrogens / Q000737:chemistry; D018517:Plant Roots / Q000033:anatomy & histology / Q000737:chemistry; D029908:Pueraria / Q000737:chemistry*; D011817:Rabbits; D012015:Reference Standards
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D048789:Phytoestrogens
ELISA; Isomiroestrol; Pueraria candollei var. mirifica; White Kwao Krua