High-Throughput Covalent Modifier Screening with Acoustic Ejection Mass Spectrometry

• DOI: 10.1021/jacs.4c02377
• Publication Date: 2024-07-12T09:30:49Z
• Authors: Xiujuan Wen; Chang Liu; Kiersten Tovar; Patrick Curran; Matthew Richards; Sony Agrawal; Richard Johnstone; Ryan E. Loy; Joey L. Methot; My Sam Mansueto; Markus Koglin; Mary Jo Wildey; Lyle Burton; Thomas R. Covey; Kevin P. Bateman; Michael Kavana; David G. McLaren
• Keywords: Small Molecule Libraries; Drug Discovery; Humans; Acoustics; Ligands; Mass Spectrometry; High-Throughput Screening Assays

Interests in covalent drugs have grown modern drug discovery as they could tackle challenging targets traditionally considered "undruggable". The identification of binders to target proteins typically involves directly measuring protein modifications using high-resolution mass spectrometry. With a continually expanding library compounds, conventional spectrometry platforms such LC-MS and SPE-MS become limiting factors for high-throughput screening. Here, we introduce prototype acoustic ejection (AEMS) system the rapid screening modifier comprising ∼10,000 compounds against 50 kDa-sized protein─Werner syndrome helicase. samples were arranged 1536-well format. sample buffer containing high-concentration salts was analyzed without any cleanup steps, minimizing preparation efforts ensuring stability. entire AEMS analysis process be completed within mere 17 h. An automated data tool facilitated batch processing quantitation formation various protein-ligand adducts. results displayed high degree fidelity, with