title: Performance of the BG1Luc ER TA method in a qHTS format.
abstract: In 2012, the BG1Luc4E2 estrogen receptor (ER) transactivation (TA) method (BG1Luc ER TA) was accepted by U.S. regulatory agencies and the Organisation for Economic Co-operation and Development to detect substances with ER agonist activity. The method is now part of the Tier 1 testing battery in the Environmental Protection Agency's Endocrine Disruptor Screening Program. The BG1Luc ER TA method uses the BG1 ovarian cell line that endogenously expresses full-length ER (α and β) and is stably transfected with a plasmid containing four estrogen responsive elements upstream of a luciferase reporter gene. To allow increased throughput and testing efficiency, the BG1Luc ER TA ("BG1 manual") method was adapted for quantitative high-throughput screening (BG1 qHTS) in the U.S. Tox21 testing program. The BG1 qHTS test method was used to test approximately 10,000 chemicals three times each, and concentration-response data (n=15) were analyzed to evaluate test method performance. The balanced accuracy of the BG1 qHTS test method (97% [32/33]) was determined by comparing results to ER TA performance standards for the BG1 manual method. Concordance between the BG1 manual and qHTS methods was 92% (57/62) when calculated for a larger set of non-reference chemicals tested in both methods. These data demonstrate that the performance of the BG1 qHTS is similar to the currently accepted BG1 manual method, thereby establishing the utility of the BG1 qHTS method for identifying ER active environmental chemicals.
authors.affiliation: Integrated Laboratory Systems, Inc., Research Triangle Park, North Carolina, USA.
authors.forename: Patricia
authors.initials: P
authors.lastname: Ceger
authors.affiliation: Integrated Laboratory Systems, Inc., Research Triangle Park, North Carolina, USA.
authors.forename: David
authors.initials: D
authors.lastname: Allen
authors.affiliation: National Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA.
authors.forename: Ruili
authors.initials: R
authors.lastname: Huang
authors.affiliation: National Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA.
authors.forename: Menghang
authors.initials: M
authors.lastname: Xia
authors.affiliation: National Toxicology Program Interagency Center for the Evaluation of Alternative Toxicological Methods, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA.
authors.forename: Warren
authors.initials: W
authors.lastname: Casey
mesh_terms: D000826:Animal Testing Alternatives; D002460:Cell Line; D052244:Endocrine Disruptors / Q000737:chemistry*; D047628:Estrogen Receptor alpha / Q000819:agonists*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D011485:Protein Binding / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014481:United States; D014484:United States Environmental Protection Agency
chemical_list: D052244:Endocrine Disruptors; D047628:Estrogen Receptor alpha
keywords: BG1Luc ER TA; EDSP; Tox21; quantitative high-throughput screening