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Journal of Biomolecular Screening
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A High-Throughput Ligand Competition Binding Assay for the Androgen Receptor and Other Nuclear Receptors

Clémentine Féau

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee

Leggy A. Arnold

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee

Aaron Kosinski

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee

R. Kiplin Guy

Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, Tennessee, kip.guy{at}stjude.org

Standardized, automated ligand-binding assays facilitate evaluation of endocrine activities of environmental chemicals and identification of antagonists of nuclear receptor ligands. Many current assays rely on fluorescently labeled ligands that are significantly different from the native ligands. The authors describe a radiolabeled ligand competition scintillation proximity assay (SPA) for the androgen receptor (AR) using Ni-coated 384-well FlashPlates® and liganded AR-LBD protein. This highly reproducible, low-cost assay is well suited for automated high-throughput screening. In addition, the authors show that this assay can be adapted to measure ligand affinities for other nuclear receptors (peroxisome proliferation-activated receptor {gamma}, thyroid receptors {alpha} and β). (Journal of Biomolecular Screening 2009:43-48)

Key Words: scintillation proximity assay • androgen receptor • high-throughput screening • endocrine disrupting chemicals • nuclear receptors

Journal of Biomolecular Screening, Vol. 14, No. 1, 43-48 (2009)
DOI: 10.1177/1087057108326662
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