HTSDSF Explorer, A Novel Tool to Analyze High-throughput DSF Screenings. Issue 11 (15th June 2022)

• Record Type: Journal Article
• Authors: Martin-Malpartida, Pau; Hausvik, Emil; Underhaug, Jarl; Torner, Carles; Martinez, Aurora; Macias, Maria J.
• Is Part Of: Journal of molecular biology. Volume 434:Issue 11(2022)
• Publication Date: 2022-06-15
• Subjects: high-throughput compound screening -- thermal shift assay -- differential scanning fluorimetry -- automatic Tm -- automatic KD analysis; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2021.167372
• Languages: English
• ISSNs: 0022-2836

Abstract

Graphical abstract: Highlights: 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. Abstract: 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 (Δ Tm ), 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 Δ Tm 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.