A1 Refereed original research article in a scientific journal
A simple heterogeneous one-step assay for screening estrogenic compounds
Authors: Huovinen T, Rytkonen K, Lamminmaki U, Pellinen T
Publisher: SPRINGER
Publication year: 2013
Journal: Biotechnology Letters
Journal name in source: BIOTECHNOLOGY LETTERS
Journal acronym: BIOTECHNOL LETT
Number in series: 1
Volume: 35
Issue: 1
First page : 47
Last page: 53
Number of pages: 7
ISSN: 0141-5492
DOI: https://doi.org/10.1007/s10529-012-1050-0
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 ER alpha. In this assay, in vivo-biotinylated (E. coli) SRC-1, purified Renilla luciferase-ER alpha, 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.
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 ER alpha. In this assay, in vivo-biotinylated (E. coli) SRC-1, purified Renilla luciferase-ER alpha, 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.
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