Samuel Errol L G, Holmes Secondra L, Young Damian W
Center for Drug Discovery, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA.
Department of Pharmacology and Chemical Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA.
J Cheminform. 2021 Dec 11;13(1):99. doi: 10.1186/s13321-021-00577-1.
The thermal shift assay (TSA)-also known as differential scanning fluorimetry (DSF), thermofluor, and T shift-is one of the most popular biophysical screening techniques used in fragment-based ligand discovery (FBLD) to detect protein-ligand interactions. By comparing the thermal stability of a target protein in the presence and absence of a ligand, potential binders can be identified. The technique is easy to set up, has low protein consumption, and can be run on most real-time polymerase chain reaction (PCR) instruments. While data analysis is straightforward in principle, it becomes cumbersome and time-consuming when the screens involve multiple 96- or 384-well plates. There are several approaches that aim to streamline this process, but most involve proprietary software, programming knowledge, or are designed for specific instrument output files. We therefore developed an analysis workflow implemented in the Konstanz Information Miner (KNIME), a free and open-source data analytics platform, which greatly streamlined our data processing timeline for 384-well plates. The implementation is code-free and freely available to the community for improvement and customization to accommodate a wide range of instrument input files and workflows.
热位移分析（TSA）——也被称为差示扫描荧光法（DSF）、热荧光法和T位移法——是基于片段的配体发现（FBLD）中用于检测蛋白质-配体相互作用的最流行的生物物理筛选技术之一。通过比较目标蛋白质在有配体和无配体存在时的热稳定性，可以识别潜在的结合剂。该技术易于设置，蛋白质消耗低，并且可以在大多数实时聚合酶链反应（PCR）仪器上运行。虽然数据分析原则上很简单，但当筛选涉及多个96孔或384孔板时，就会变得繁琐且耗时。有几种方法旨在简化这一过程，但大多数都涉及专有软件、编程知识，或者是为特定仪器输出文件设计的。因此，我们开发了一种在康斯坦茨信息挖掘器（KNIME）中实现的分析工作流程，KNIME是一个免费的开源数据分析平台，它大大简化了我们对384孔板的数据处理时间线。该实现无需编码，社区可免费使用，以便改进和定制，以适应广泛的仪器输入文件和工作流程。
