title: Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody.
abstract: Miroestrol is a chromene with potent estrogenic activity present inPueraria candollei, commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action ofP. candolleiproducts. As a tool for further studies about the efficacy and safety ofP. candolleias a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes ofP. candollei. The developed ELISA against miroestrol has a calibration range of 10-780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root ofP. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.|Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Tharita
authors.initials: T
authors.lastname: Kitisripanya
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.|Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Supaluk
authors.initials: S
authors.lastname: Krittanai
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.|Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Orapin
authors.initials: O
authors.lastname: Udomsin
authors.affiliation: Regional Medical Sciences Center 7, Department of Medical Sciences, Khon Kaen, Thailand.
authors.forename: Kamonthip
authors.initials: K
authors.lastname: Jutathis
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.|Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Jukrapun
authors.initials: J
authors.lastname: Komaikul
authors.affiliation: Faculty of Pharmaceutical Sciences, UbonRatchathani University, Ubon Ratchathani, Thailand.
authors.forename: Thaweesak
authors.initials: T
authors.lastname: Juengwatanatrakul
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Sakamoto
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Morimoto
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand.|Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D048789:Phytoestrogens / Q000032:analysis* / Q000276:immunology; D029908:Pueraria / Q000737:chemistry*; D011786:Quality Control; D013256:Steroids / Q000032:analysis* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D048789:Phytoestrogens; D013256:Steroids; C571572:miroestrol