Fast affinity purification coupled with mass spectrometry for identifying organophosphate labeled plasma butyrylcholinesterase |
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Authors: | Li He Tong Larry Schopfer Lawrence M Masson Patrick Lockridge Oksana |
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Institution: | Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-6805, United States. |
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Abstract: | Classical plasma butyrylcholinesterase (BChE) purification involves dialysis and multiple steps of chromatography. We describe a procainamide affinity gel purification scheme that takes 15-30min to purify BChE from 1ml plasma. The method uses a microfuge spin column to build a 0.2ml procainamide affinity column. The eluted BChE contains 3-4mug 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. |
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Keywords: | Butyrylcholinesterase Biomarker Organophosphate exposure Sarin Soman Affinity chromatography Mass spectrometry |
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