HTSDSF Explorer, A Novel Tool to Analyze High-throughput DSF Screenings

• Published in: Journal of molecular biology Vol. 434; no. 11; p. 167372
• Main Authors: Martin-Malpartida, Pau, Hausvik, Emil, Underhaug, Jarl, Torner, Carles, Martinez, Aurora, Macias, Maria J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 15.06.2022
• Subjects: automatic KD analysis; automatic Tm; differential scanning fluorimetry; high-throughput compound screening; thermal shift assay; automatic KD analysis; automatic Tm; high-throughput compound screening; thermal shift assay; differential scanning fluorimetry; automatic K(D) analysis; automatic T(m)
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2021.167372

[Display omitted] •A high-throughput focused DSF data analyzer, which runs very fast and in a highly efficient manner.•The software integrates plate-screening with protein–ligand affinity calculations.•Provides a Rank of all binders ordered according to differences in melting temperatures.•Generates Excel files and final KD reports automatically. The identification of new drugs for novel therapeutic targets requires the screening of libraries containing tens of thousands of compounds. While experimental screenings are assisted by high-throughput technologies, in target-based biophysical assays, such as differential scanning fluorimetry (DSF), the analysis steps must be calculated manually, often combining several software packages. To simplify the determination of the melting temperature (Tm) of the target and the change induced by ligand binding (ΔTm), we developed the HTSDSF explorer, a versatile, all-in-one, user-friendly application suite. Implemented as a server-client application, in the primary screenings, HTSDSF explorer pre-analyzes and displays the Tm and ΔTm results interactively, thereby allowing the user to study hundreds of conditions and select the primary hits in minutes. This application also allows the determination of preliminary binding constants (KD) through a series of subsequent dose–response assays on the primary hits, thereby facilitating the ranking of validated hits and the advance of drug discovery efforts.