Fast affinity purification coupled with mass spectrometry for identifying organophosphate labeled plasma butyrylcholinesterase

He Li, Larry Tong, Lawrence M. Schopfer, Patrick Masson, Oksana Lockridge

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

Classical plasma butyrylcholinesterase (BChE) purification involves dialysis and multiple steps of chromatography. We describe a procainamide affinity gel purification scheme that takes 15-30 min to purify BChE from 1 ml plasma. The method uses a microfuge spin column to build a 0.2 ml procainamide affinity column. The eluted BChE contains 3-4 μg of 500-fold purified BChE, free from 99% of contaminating plasma proteins. The BChE was further purified by gel electrophoresis. Tryptic peptides from the BChE containing gel electrophoresis band were prepared by in-gel digestion, separated by reverse phase liquid chromatography and identified by mass spectrometry. The 29 residue active site tryptic peptide labeled with the nerve agents soman or sarin was identified.

Original languageEnglish (US)
Pages (from-to)68-72
Number of pages5
JournalChemico-Biological Interactions
Volume175
Issue number1-3
DOIs
StatePublished - Sep 25 2008

Keywords

  • Affinity chromatography
  • Biomarker
  • Butyrylcholinesterase
  • Mass spectrometry
  • Organophosphate exposure
  • Sarin
  • Soman

ASJC Scopus subject areas

  • Toxicology

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