High-throughput Screening, Discovery Sciences, R&D, AstraZeneca, Alderley Park, UK.
Discovery Biology, Discovery Sciences, R&D, AstraZeneca, Cambridge, UK.
SLAS Discov. 2023 Apr;28(3):102-110. doi: 10.1016/j.slasd.2023.01.006. Epub 2023 Feb 1.
The cellular thermal shift assay (CETSA®) has increasingly been used in early drug discovery to provide a measure of cellular target engagement. Traditionally, CETSA has been employed for bespoke questions with small to medium throughput and has predominantly been applied during hit validation rather than in hit identification. Using a CETSA screen versus the kinase CRAF, we assessed 3 key questions: (1) technical feasibility - could the CETSA methodology technically be applied at truly high throughput scale? (2) relevance - could hits suitable for further optimisation be identified? (3) reliability - would the approach identify known chemical equity. Here, we describe the first large scale AlphaLISA SureFire based CETSA (Alpha CETSA) approach allowing us to screen a large library of almost 0.5 million compounds. We discuss the issues overcome in automating and executing the screen and describe the resulting screen output.
细胞热转移分析（CETSA®）已越来越多地用于早期药物发现，以提供细胞靶标结合的衡量标准。传统上，CETSA 用于具有小到中等通量的定制问题，主要应用于命中验证而不是命中鉴定。我们使用 CETSA 筛选与激酶 CRAF 的对比，评估了 3 个关键问题：（1）技术可行性- CETSA 方法在真正的高通量规模上是否可以在技术上应用？（2）相关性-是否可以确定适合进一步优化的命中？（3）可靠性-该方法是否可以识别已知的化学权益。在这里，我们描述了第一个基于 AlphaLISA SureFire 的大规模 CETSA（Alpha CETSA）方法，使我们能够筛选近 50 万种化合物的大型文库。我们讨论了在自动化和执行筛选中克服的问题，并描述了由此产生的筛选输出。
