High Throughput Screening with SAMDI Mass Spectrometry for Directed Evolution

Adam J. Pluchinsky, Daniel J. Wackelin, Xiongyi Huang, Frances H. Arnold, Milan Mrksich*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Advances in directed evolution have led to an exploration of new and important chemical transformations; however, many of these efforts still rely on the use of low-throughput chromatography-based screening methods. We present a high-throughput strategy for screening libraries of enzyme variants for improved activity. Unpurified reaction products are immobilized to a self-assembled monolayer and analyzed by mass spectrometry, allowing for direct evaluation of thousands of variants in under an hour. The method was demonstrated with libraries of randomly mutated cytochrome P411 variants to identify improved catalysts for C-H alkylation. The technique may be tailored to evolve enzymatic activity for a variety of transformations where higher throughput is needed.

Original languageEnglish (US)
Pages (from-to)19804-19808
Number of pages5
JournalJournal of the American Chemical Society
Volume142
Issue number47
DOIs
StatePublished - Nov 25 2020

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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