Lee, P.-H., Huang, X. X., Teh, B. T., & Ng, L.-M. (2019). TSA-CRAFT: A Free Software for Automatic and Robust Thermal Shift Assay Data Analysis. SLAS DISCOVERY: Advancing the Science of Drug Discovery, 24(5), 606–612. https://doi.org/10.1177/2472555218823547
Thermal shift assay (TSA) is an increasingly popular technique used for identifying protein stabilizing conditions or interacting ligands in X-ray crystallography and drug discovery applications. Although the setting up and running of TSA reactions is a relatively simple process, the subsequent analysis of TSA data, especially in high-throughput format, requires substantial amount of effort if conducted manually. We therefore developed the Thermal Shift Assay–Curve Rapid and Automatic Fitting Tool (TSA-CRAFT), a freely available software that enable automatic analysis of TSA data of any throughput. TSA-CRAFT directly reads real-time PCR instrument data files and displays the analyzed results in a web browser. This software features streamlined data processing and Boltzmann equation fitting, which is demonstrated in this study to provide more accurate data analysis than the commonly used first-derivative method. TSA-CRAFT is freely available as a cross-operating system-compatible standalone tool (https://sourceforge.net/projects/tsa-craft/) and also as a freely accessible web server (http://tbtlab.org/tsacraft.html).
This work is funded by National Medical Research Council (NMRC OFIRG16may055) and supported by the National Research Foundation Singapore and the Singapore Ministry of Education under its Research Centres of Excellence initiative.