Download PDF
Planta Med 2017; 83(10): 855-861
DOI: 10.1055/s-0043-102689
Natural Product Chemistry and Analytical Studies
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

Tharita Kitisripanya
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Supaluk Krittanai
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Orapin Udomsin
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Kamonthip Jutathis
3   Regional Medical Sciences Center 7, Department of Medical Sciences, Khon Kaen, Thailand
,
Jukrapun Komaikul
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
,
Thaweesak Juengwatanatrakul
4   Faculty of Pharmaceutical Sciences, UbonRatchathani University, Ubon Ratchathani, Thailand
,
Seiichi Sakamoto
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Hiroyuki Tanaka
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Satoshi Morimoto
5   Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan
,
Waraporn Putalun
1   Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, Thailand
2   Research Group for Pharmaceutical Activities of Natural Products Using Pharmaceutical Biotechnology (PANPB), National Research University – Khon Kaen University, Khon Kaen, Thailand
› Author Affiliations
Further Information

Publication History

received 31 October 2016
revised 29 December 2016

accepted 25 January 2017

Publication Date:
10 February 2017 (online)

Also available at
    Permissions and Reprints

Abstract

Miroestrol is a chromene with potent estrogenic activity present in Pueraria 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 of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as 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 of P. 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 of P. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

Key words

miroestrol - monoclonal antibody - enzyme-linked immunosorbent assay (ELISA) - immunoaffinity column - White Kwao Krua - Pueraria candollei - Fabaceae
 

  • References

  • 1 Malaivijitnond S. Medical applications of phytoestrogens from the Thai herb Pueraria mirifica . Front Med 2012; 6: 8-21
    CrossrefPubMedGoogle Scholar
  • 2 Suntara A. The Remedy Pamphlet of Kwao Krua Tuber of Luang Anusarnsuntarakromkarnphiset. 1st ed. Chiang Mai: Upatipongsa Press; 1931
    Google Scholar
  • 3 Urasopon N, Hamada Y, Asaoka K, Poungmali U, Malaivijitnond S. Isoflavone content of rodent diets and its estrogenic effect on vaginal cornification in Pueraria mirifica-treated rats. Sci Asia 2008; 34: 371-376
    CrossrefPubMedGoogle Scholar
  • 4 Lee YS, Park JS, Cho SD, Son JK, Cherdshewasart W, Kang KS. Requirement of metabolic activation for estrogenic activity of Pueraria mirifica . J Vet Sci 2002; 3: 273-277
    CrossrefPubMedGoogle Scholar
  • 5 Trisomboon H, Malaivijitnond S, Watanabe G, Taya K. Estrogenic effects of Pueraria mirifica on the menstrual cycle and hormone-related ovarian functions in cyclic female cynomolgus monkeys. J Pharmacol Sci 2004; 94: 51-59
    CrossrefPubMedGoogle Scholar
  • 6 Chatuphonprasert W, Udomsuk L, Monthakantirat O, Churikhit Y, Putalun W, Jarukamjorna K. Effects of Pueraria mirifica and miroestrol on the antioxidation-related enzymes in ovariectomized mice. J Pharm Pharmacol 2013; 65: 447-456
    CrossrefPubMedGoogle Scholar
  • 7 Urasopon N, Hamada Y, Cherdshewasart W, Malaivijitnond S. Preventive effects of Pueraria mirifica on bone loss in ovariectomized rats. Maturitas 2008; 59: 137-148
    CrossrefPubMedGoogle Scholar
  • 8 Udomsuk L, Chatuphonprasert W, Monthakantirat O, Churikhit Y, Jarukamjorn K. Impact of Pueraria candollei var. mirifica and its potent phytoestrogen miroestrol on expression of bone-specific genes in ovariectomized mice. Fitoterapia 2012; 83: 1687-1692
    CrossrefPubMedGoogle Scholar
  • 9 Van der Maesen LJG. Pueraria: botanical Characteristics. In: Keung WM. ed. Pueraria: The Genus Pueraria . London: Taylor & Francis; 2002: 1-28
    Google Scholar
  • 10 Chandeying V, Lamlertkittikul S. Challenges in the conduct of Thai herbal scientific study: efficacy and safety of phytoestrogen, Pueraria mirifica (Kwao Keur Kao), phase I, in the alleviation of climacteric symptoms in perimenopausal women. J Med Assoc Thai 2007; 90: 1274-1280
    PubMedGoogle Scholar
  • 11 Lamlertkittikul S, Chandeying V. Efficacy and safety of Pueraria mirifica (Kwao Kruea Khao) for the treatment of vasomotor symptoms in perimenopausal women: Phase II study. J Med Assoc Thai 2004; 87: 33-40
    PubMedGoogle Scholar
  • 12 Manosroi A, Manosroi J. Determination of bioactive compounds in roots of different ages Pueraria mirifica, airy shaw suvatabhandhu and Butea superba, roxb. from various locations in Thailand. Acta Hortic 2005; 680: 93-96
    PubMedGoogle Scholar
  • 13 Pongkitwitoon B, Sakamoto S, Tanaka H, Tsuchihashi R, Kinjo J, Morimoto S, Putalun W. Enzyme-linked immunosorbent assay for total isoflavonoids in Pueraria candollei using anti-puerarin and anti-daidzin polyclonal antibodies. Planta Med 2010; 76: 831-836
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Matsumura A, Ghosh A, Pope GS, Darbre PD. Comparative study of oestrogenic properties of eight phytoestrogens in MCF7 human breast cancer cells. J Steroid Biochem Mol Biol 2005; 94: 431-443
    CrossrefPubMedGoogle Scholar
  • 15 Chansakaow S, Ishikawa T, Seki H, Sekine K, Okada M, Chaichantipyuth C. (née Yoshizawa) Identification of deoxymiroestrol as the actual rejuvenating principle of “Kwao Keur”, Pueraria mirifica. The known miroestrol may be an artifact. J Nat Prod 2000; 63: 173-175
    CrossrefPubMedGoogle Scholar
  • 16 Udomsuk L, Juengwattanatrakul T, Jarukamjorn K, Putalun W. Increased miroestrol, deoxymiroestrol and isoflavonoid accumulation in callus and cell suspension cultures of Pueraria candollei var. mirifica . Acta Physiol Plant 2012; 34: 1093-1100
    CrossrefPubMedGoogle Scholar
  • 17 Shimokawa S, Kumamoto T, Ishikawa T, Takashi M, Higuchi Y, Chaichantipyuth C, Chansakaow S. Quantitative analysis of miroestrol and kwakhurin for standardisation of Thai miracle herb ʼKwao Keurʼ (Pueraria mirifica) and establishment of simple isolation procedure for highly estrogenic miroestrol and deoxymiroestrol. Nat Prod Res 2013; 27: 371-378
    CrossrefPubMedGoogle Scholar
  • 18 Yusakul G, Putalun W, Udomsin O, Juengwatanatrakul T, Chaichantipyuth C. Comparative analysis of the chemical constituents of two varieties of Pueraria candollei . Fitoterapia 2011; 82: 203-207
    CrossrefPubMedGoogle Scholar
  • 19 Yusakul G, Udomsin O, Juengwatanatrakul T, Tanaka H, Chaichantipyuth C, Putalun W. High performance enzyme-linked immunosorbent assay for determination of miroestrol, a potent phytoestrogen from Pueraria candollei . Anal Chim Acta 2013; 785: 104-110
    CrossrefPubMedGoogle Scholar
  • 20 Yusakul G, Udomsin O, Tanaka H, Morimoto S, Juengwatanatrakul T, Putalun W. Enzyme-linked immunosorbent assay by enhanced chemiluminescence detection for the standardization of estrogenic miroestrol in Pueraria candollei Graham ex Benth. Luminescence 2015; 30: 568-575
    CrossrefPubMedGoogle Scholar
  • 21 Kitisripanya T, Jutathis K, Inyai C, Komaikul J, Udomsin O, Yusakul G, Tanaka H, Putalun W. Anti-miroestrol polyclonal antibodies: a comparison of immunogen preparations used to obtain desired antibody properties. J Nat Med 2016; 70: 296-299
    CrossrefPubMedGoogle Scholar
  • 22 Moser AC, Hage DS. Immunoaffinity chromatography: an introduction to applications and recent developments. Bioanalysis 2010; 2: 769-790
    CrossrefPubMedGoogle Scholar
  • 23 Yanagihara H, Sakata R, Minami H, Tanaka H, Shoyama Y, Murakami H. Immunoaffinity column chromatography against forskolin using an anti-forskolin monoclonal antibody and its application. Anal Chim Acta 1996; 335: 63-70
    CrossrefPubMedGoogle Scholar
  • 24 Putalun W, Tanaka H, Yukihira S. Rapid separation of solasodine glycosides by an immunoaffinity column using anti-solamargine monoclonal antibody. Cytotechnology 1999; 31: 151-156
    PubMedGoogle Scholar
  • 25 Uto T, Tung NH, Morinaga O, Shoyama Y. Preparation of knockout extract by immunoaffinity column and its application. Antibodies (Basel) 2012; 1: 294-307
    CrossrefPubMedGoogle Scholar
  • 26 Qu H, Zhang G, Li Y, Sun H, Sun Y, Zhao Y, Wang Q. Development of an enzyme-linked immunosorbent assay based on anti-puerarin monoclonal antibody and its applications. J Chromatogr B Analyt Technol Biomed Life Sci 2014; 953–954: 120-125
    PubMedGoogle Scholar
  • 27 Pongkitwitoon B, Sakamoto S, Tanaka H, Tsuchihashi R, Kinjo J, Morimoto S, Putalun W. Development of an enzyme-linked immunosorbent assay to determine puerarin and its aglycone daidzein. J Nat Med 2011; 65: 31-36
    CrossrefPubMedGoogle Scholar
  • 28 OʼFegan PO. Validation. In: Gosling JP. ed. Immunoassays. New York: Oxford University Press; 2000: 211-238
    Google Scholar
  • 29 Muckerheide A, Apple RJ, Pesce AJ, Michael JG. Cationization of protein antigens. I. Alteration of immunogenic properties. J Immunol 1987; 138: 833-837
    PubMedGoogle Scholar
  • 30 Fuller SA, Takahashi M, Hurrell JG. Fusion of myeloma cells with immune spleen cells. Curr Protoc Mol Biol 2001; Chapter 11: Unit 11.7
    PubMedGoogle Scholar
  • 31 Fuller SA, Takahashi M, Hurrell JG. Cloning of hybridoma cell lines by limiting dilution. Curr Protoc Mol Biol 2001; Chapter 11: Unit 11.8
    PubMedGoogle Scholar
  • 32 Udomsin O, Juengwatanatrakul T, Yusakul G, Putalun W. Chromene stability: The most potent estrogenic compounds in White Kwao Krua (Pueraria candollei var. mirifica) crude extract. J Funct Foods 2015; 19A: 269-277
    PubMedGoogle Scholar