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Screening for androgen agonists using autonomously bioluminescent HEK293 reporter cells.
Biotechniques ( IF 2.2 ) Pub Date : 2021-08-05 , DOI: 10.2144/btn-2021-0017
Tingting Xu 1 , Madison Gilliam 2 , Gary Sayler 1, 3 , Steven Ripp 1, 3 , Dan Close 3
Affiliation  

Due to the public health concerns of endocrine-disrupting chemicals, there is an increasing demand to develop improved high-throughput detection assays for enhanced exposure control and risk assessment. A substrate-free, autobioluminescent HEK293ARE/Gal4-Lux assay was developed to screen compounds for their ability to induce androgen receptor (AR)-mediated transcriptional activation. The assay was validated against a group of 40 recommended chemicals and achieved an overall 87.5% accuracy in qualitatively classifying positive and negative AR agonists. The HEK293ARE/Gal4-Lux assay was demonstrated as a suitable tool for Tier 1 AR agonist screening. By eliminating exogenous substrate, this assay provided a significant advantage over traditional reporter assays by enabling higher-throughput screening with reduced testing costs while maintaining detection accuracy.

中文翻译:

使用自主生物发光 HEK293 报告细胞筛选雄激素激动剂。

由于内分泌干扰化学品的公共卫生问题,越来越需要开发改进的高通量检测分析,以加强接触控制和风险评估。开发了一种无底物、自体发光 HEK293 ARE/Gal4-Lux测定法来筛选化合物诱导雄激素受体 (AR) 介导的转录激活的能力。该测定针对一组 40 种推荐的化学物质进行了验证,在对阳性和阴性 AR 激动剂进行定性分类方面达到了 87.5% 的总体准确率。HEK293 ARE/Gal4-Lux试验被证明是用于 Tier 1 AR 激动剂筛选的合适工具。通过消除外源性底物,该检测提供了优于传统报告检测的显着优势,可实现更高通量的筛选,同时降低测试成本,同时保持检测准确性。
更新日期:2021-08-05
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