Related Experiment Video
Updated: May 18, 2026
Screening for Phytoestrogens using a Cell-based Estrogen Receptor β Reporter Assay
Published on: June 7, 2020
A simple heterogeneous one-step assay for screening estrogenic compounds
Tuomas Huovinen1, Kalle Rytkönen, Urpo Lamminmäki
1Department of Biotechnology, University of Turku, Turku, Finland. tuomas.huovinen@utu.fi
Abstract:
Estrogen receptor (ER) modulators are a serious health issue but estrogenic compounds, especially antagonists of ER function, are widely screened for in search of novel therapeutics against hormonal diseases such as the breast cancer. Here we report a novel and a simple bioassay for estrogenic and anti-estrogenic compounds based on ligand-dependent recruitment of ER co-activator steroid receptor co-activator 1 (SRC-1) to purified Renilla luciferase-tagged ERα. In this assay, in vivo-biotinylated (E. coli) SRC-1, purified Renilla luciferase-ERα, and the analyte sample are mixed and incubated for 2 h in a streptavidin-coated microtiter wells, and after one washing step, luminescence is measured with a simple instrument. The assay does not require chemical labeling of the components and shows good sensitivity (25 pM E(2)) and wide dynamic range of more than four orders of magnitude.
Insights
A new bioassay detects estrogenic and anti-estrogenic compounds by measuring co-activator recruitment to estrogen receptor alpha (ERα). This simple assay aids in discovering therapeutics for hormonal diseases like breast cancer.
Area of Science:
- Biochemistry
- Molecular Biology
- Endocrinology
Background:
- Estrogen receptor (ER) modulators pose health risks, yet ER antagonists are crucial for developing therapeutics against hormonal diseases, including breast cancer.
- Screening for novel estrogenic and anti-estrogenic compounds is vital for advancing treatments for hormone-dependent conditions.
Purpose of the Study:
- To develop a novel, simple bioassay for detecting estrogenic and anti-estrogenic compounds.
- To utilize ligand-dependent co-activator recruitment for ERα activity assessment.
Main Methods:
- The assay employs purified Renilla luciferase-tagged ERα and in vivo-biotinylated steroid receptor co-activator 1 (SRC-1).
- Compounds are incubated with ERα and SRC-1 in streptavidin-coated microtiter wells, followed by a washing step and luminescence measurement.
- The method avoids chemical labeling of assay components.
Main Results:
- The bioassay demonstrates high sensitivity, detecting estradiol (E2) at concentrations as low as 25 pM.
- The assay exhibits a wide dynamic range, spanning over four orders of magnitude.
- Ligand-dependent recruitment of SRC-1 to ERα is effectively measured.
Conclusions:
- This novel bioassay provides a simple and sensitive method for screening estrogenic and anti-estrogenic compounds.
- The assay's capabilities support the search for new therapeutics targeting hormonal diseases.
- The method's efficiency and broad dynamic range make it valuable for drug discovery efforts.
More Related Videos
14:13Detecting Estrogenic Ligands in Personal Care Products using a Yeast Estrogen Screen Optimized for the Undergraduate Teaching Laboratory
Published on: January 1, 2018
08:00Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays
Published on: December 4, 2016
