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A nonisotopic estrogen receptor–based assay to detect estrogenic compounds

Nature Biotechnology volume 17, pages 1219–1222 (1999)Cite this article

Abstract

We have used the ligand binding domain of the recombinant human estrogen receptor (hER) to develop a nonisotopic assay for detection of estrogenic compounds. The assay is based on competition of the estrogenic ligand with 17β-estradiol for binding to the receptor, which leaves 17β-estradiol free to bind to an anti–17β-estradiol antibody. Unbound anti–17β-estradiol antibody then binds to immobilized 17β-estradiol–protein conjugate (to which hER is unable to bind for steric reasons), and is detected by an enzyme-labeled anti-rabbit IgG antibody. We used the assay to detect estrogenic compounds (mainly members of the flavonoid group of plant polyphenols) in a variety of commonly consumed plant foods.

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Figure 1: Competition of various estrogenic compounds with [3H]–17β–estradiol for the hER–LBD.
Figure 2
Figure 3: Diagrammatic representation of the receptor–based microtitration plate assay.
Figure 4: Competitive binding curves of various estrogenic compounds in the nonisotopic receptor–based assay.

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Acknowledgements

The financial support of the Ministry of Agriculture, Fisheries, and Food is gratefully acknowledged. We thank Elif Buyukpamukcu for making the 17β–estradiol–BTG conjugate; Geoffrey Greene of the Ben May Institute, University of Chicago for providing the GST–LBD plasmid; Jason Liggins of the Dunn Nutrition Centre, University of Cambridge, UK, for helpful discussions and isoflavone measurements of some foods; Keith Price of the Institute of Food Research (Norwich, UK) for valuable discussions on flavonoids in foods; and Hugh Makin of Queen Mary & Westfield College, University of London, London, UK, for helpful discussions and his support.

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  1. Institute of Food Research, Norwich Research Park, Colney, NR4 7UA, Norwich, United Kingdom

    Stephen D. Garrett, Heather A. Lee & Michael R.A. Morgan

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  1. Stephen D. Garrett
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Correspondence to Heather A. Lee.

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Garrett, S., Lee, H. & Morgan, M. A nonisotopic estrogen receptor–based assay to detect estrogenic compounds. Nat Biotechnol 17, 1219–1222 (1999). https://doi.org/10.1038/70773

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