Drug Target Engagement Using Coupled Cellular Thermal Shift Assay—Acoustic Reverse-Phase Protein Array. (February 2020)

• Record Type: Journal Article
• Authors: Herledan, Adrien; Andres, Marine; Lejeune-Dodge, Aurore; Leroux, Florence; Biela, Alexandre; Piveteau, Catherine; Warenghem, Sandrine; Couturier, Cyril; Deprez, Benoit; Deprez-Poulain, Rebecca
• Is Part Of: SLAS discovery. Volume 25:Number 2(2020)
• Page Start: 207
• Page End: 214
• Publication Date: 2020-02
• Subjects: target engagement -- CETSA -- RPPA -- acoustic transfer -- drug discovery -- nanovolume -- insulin-degrading enzyme; 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: 10.1177/2472555219897256
• Languages: English
• ISSNs: 2472-5552

Abstract

In the last 5 years, cellular thermal shift assay (CETSA), a technology based on ligand-induced changes in protein thermal stability, has been increasingly used in drug discovery to address the fundamental question of whether drug candidates engage their intended target in a biologically relevant setting. To analyze lysates from cells submitted to increasing temperature, the detection and quantification of the remaining soluble protein can be achieved using quantitative mass spectrometry, Western blotting, or AlphaScreen techniques. Still, these approaches can be time- and cell-consuming. To cope with limitations of throughput and protein amount requirements, we developed a new coupled assay combining the advantages of a nanoacoustic transfer system and reverse-phase protein array technology within CETSA experiments. We validated the technology to assess engagement of inhibitors of insulin-degrading enzyme (IDE), an enzyme involved in diabetes and Alzheimer's disease. CETSA—acoustic reverse-phase protein array (CETSA-aRPPA) allows simultaneous analysis of many conditions and drug–target engagement with a small sample size, in a rapid, cost-effective, and biological material-saving manner.