High-Throughput Mass Spectrometry in Drug Discovery

SLAS TECHNOLOGY, Journal Year: 2025, Volume and Issue: unknown, P. 100292 - 100292

Published: April 1, 2025

Language: Английский

Direct Microplate Sampling Mass Spectrometry for High-Throughput Screening of Biocatalytic Activity

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: May 15, 2025

The development of biocatalysis depends heavily on high-throughput screening (HTS) approaches to uncover engineered enzymes with superior biocatalytic activity. Recent advances in mass spectrometry (MS) enable label-free, high-speed analysis samples standard microplates by omitting the chromatographic separation. However, most MS methods need a complicated sampling interface system and face challenges due biological matrix interferences, which can diminish sensitivity reliability. Herein, we established direct microplate (DMS)-MS technique for HTS enzymatic activity 17β-hydroxysteroid dehydrogenase (17β-HSD) steroid biocatalysis. implementation an open port (OPI) probe customized ion source microwell insertion ionization has streamlined setup, reduced operational complexity, shortened time 0.9-6.5 s/sample. A sample preparation strategy involving derivatization, followed high-ratio dilution (e.g., 10,000-fold), was further integrated prior DMS-MS mitigating suppression effects across diverse matrices. Ultimately, this workflow, when applied monitoring multiple steroids 17β-HSD processes, demonstrated high stability (peak area RSDs < 7%), minimal carryover (<1%), quantitative accuracy comparable conventional liquid chromatography (LC) experiments while providing >150-fold throughput enhancement. After total 5760 mutants three different conversion reactions, identified verified two new variants enhanced activities. This study successfully DMS-MS-based workflow screening, simplified, robust, highly reliable analytical platform microbial or enzyme engineering projects complex

Language: Английский

Notes on AEMS Methods Development for High Throughput Experimentation in Drug Discovery.

SLAS TECHNOLOGY, Journal Year: 2024, Volume and Issue: unknown, P. 100234 - 100234

Published: Dec. 1, 2024

Language: Английский