TSA-CRAFT: A Free Software for Automatic and Robust Thermal Shift Assay Data Analysis. (June 2019)

• Record Type: Journal Article
• Authors: Lee, Po-Hsien; Huang, Xi Xiao; Teh, Bin Tean; Ng, Ley-Moy
• Is Part Of: SLAS discovery. Volume 24:Number 5(2019)
• Page Start: 606
• Page End: 612
• Publication Date: 2019-06
• Subjects: thermal shift assay -- thermofluor -- differential scanning fluorimetry -- automatic Tm analysis -- high-throughput ligand screening; Drugs -- Analysis -- Periodicals; Drugs -- Testing -- Periodicals; Biomolecules -- Analysis -- Periodicals; Biomolecules -- Analysis; Drugs -- Analysis; Drugs -- Testing; Drug Evaluation, Preclinical; Molecular Biology -- methods; Periodicals; Periodicals; 615.1
• DOI: 10.1177/2472555218823547
• Languages: English
• ISSNs: 2472-5552

Abstract

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 ).