Abstract：
The Cellular Thermal Shift Assay (CETSA) enables the study of protein-ligand interactions in a cellular context. It provides valuable information on the binding affinity and specificity of both small and large molecule ligands in a relevant physiological context, hence forming a unique tool in drug discovery. Though high-throughput lab protocols exist for scaling up CETSA, subsequent data analysis and quality control remain laborious and limit experimental throughput. Here, we introduce a scalable and robust data analysis workflow which allows integration of CETSA into routine high throughput screening (HT-CETSA). This new workflow automates data analysis and incorporates quality control (QC), including outlier detection, sample and plate QC, and result triage. We describe the workflow and show its robustness against typical experimental artifacts, show scaling effects, and discuss the impact of data analysis automation by eliminating manual data processing steps.
摘要：
细胞热移位测定(CETSA)使得能够研究细胞环境中的蛋白质-配体相互作用。它提供了有关小分子和大分子配体在相关生理环境中的结合亲和力和特异性的有价值的信息，从而形成了药物发现的独特工具。尽管存在用于扩展CETSA的高通量实验室协议，随后的数据分析和质量控制仍然很费力，限制了实验通量。这里,我们引入了可扩展且稳健的数据分析工作流程,该流程允许将CETSA整合到常规高通量筛查(HT-CETSA)中.这个新的工作流程自动化数据分析，并纳入质量控制（QC），包括异常值检测，样品和板QC，和结果分类。我们描述了工作流程，并展示了它对典型实验工件的鲁棒性，显示缩放效果，并讨论了通过消除手动数据处理步骤来实现数据分析自动化的影响。
