A nonisotopic estrogen receptor−based assay to detect estrogenic
compounds
Stephen D. Garrett, Heather A. Lee
& Michael R.A. Morgan
Institute of Food Research, Norwich Research Park,
Colney, Norwich, NR4 7UA, United Kingdom.
Correspondence should be addressed to Heather A. Lee heather.lee@bbsrc.ac.ukhuman estrogen receptorphytoestrogensestrogensrapid screeningflavonoids
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
17-estradiol for binding to the receptor, which leaves 17-estradiol
free to bind to an anti−17-estradiol antibody. Unbound anti−17-estradiol
antibody then binds to immobilized 17-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.
-estradiol for binding to the receptor, which leaves 17
