Ligand competition binding assay for the androgen receptor

Area of Science:

• Endocrinology and Pharmacology
• Molecular Biology and Biochemistry

Background:

• Accurate evaluation of endocrine-disrupting chemicals requires robust androgen receptor (AR) assays.
• Standardized procedures are essential for screening environmental chemicals and identifying novel AR modulators.

Purpose of the Study:

• To develop and validate a standardized, reliable, and cost-effective assay for AR ligand competition binding.
• To adapt the assay for high-throughput screening (HTS) applications.

Main Methods:

• Utilized a radiolabeled ligand competition binding assay.
• Implemented the assay in a 384-well format.
• Automated the assay for high-throughput screening (HTS).

Main Results:

• Developed a highly reproducible scintillation proximity assay (SPA) for AR ligand binding.
• The assay is suitable for screening small molecule modulators and environmental chemicals.
• Demonstrated the assay's suitability for automation and HTS.

Conclusions:

• The developed SPA provides a standardized, reliable, and low-cost method for AR ligand binding assays.
• This assay facilitates efficient screening for endocrine activity and identification of AR modulators.
• Automation enables large-scale high-throughput screening (HTS) critical for environmental and drug discovery efforts.