6/29/2023 0 Comments Millipore quickscale 70 x 550Purified BChE is stable for up to 20 years when a sterile, concentrated solution (e.g. Animals pretreated with purified BChE are protected from lethal doses of soman, sarin, and VX with no signs of toxicity, cognitive deficits, or behavioral decrements in comparison to animals not pretreated with human BChE.īChE purified from human plasma is a tetramer with a half-life in the circulation of 2 weeks. Human butyrylcholinesterase (BChE) in blood protects from the toxicity of nerve agents and organophosphorus pesticides by inactivating these poisons before they reach the nervous system. Prekallikrein activator, Hageman Factor, coagulation Factor XII, accession P00748 S/D, Phosphate buffered saline (12 mM sodium/potassium phosphate pH 7.4 plus 13 mM sodium chloride, 2.5 mM potassium chloride) PK, Liquid chromatography tandem mass spectrometry PBS, or French government agencies.Ĭompeting interests: The authors have declared that no competing interests exist.īutyrylcholinesterase, accession P06276 LC-MS/MS, Use of trade names is for identification only and does not imply endorsement by U.S. Defense Threat Reduction Agency or the U.S. The findings and conclusions in this article are those of the authors and do not necessarily represent the views of the U.S. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Emilie David, President of the CHEMFORASE Company, synthesized Hupresin. CHEMFORASE thanks The French Ministry of Higher Education and Research, Bpifrance, Normandie Incubation and Réseau Entreprendre Normandie Seine for financial support URL =. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: This work was supported by contract HDTRA1-14-1-0056 (to SH) from the United States Defense Threat Reduction Agency URL = and by the Fred & Pamela Buffet Cancer Center Support Grant P30CA036727 from NIH URL =. Received: SeptemAccepted: DecemPublished: January 13, 2023Ĭopyright: © 2023 M. PLoS ONE 18(1):Įditor: Ajaya Bhattarai, Tribhuvan University, NEPAL Schopfer L, David E, Hinrichs SH, Lockridge O (2023) Human butyrylcholinesterase in Cohn fraction IV-4 purified in a single chromatography step on Hupresin. The new method significantly reduces production costs by optimizing yield of 99% pure butyrylcholinesterase.Ĭitation: M. This new, 1-step chromatography protocol on Hupresin increased the yield of butyrylcholinesterase by 200%. The chromogenic assay indicated kallikrein activity was less than 9 mU/mL. Prekallikrein and kallikrein were not detected by mass spectrometry in the 99% pure BChE. Possible contamination by prekallikrein and prekallikrein activator was examined by LC-MS/MS and by a chromogenic substrate assay for kallikrein activity. A high pump rate produced protein aggregates, but a low pump rate caused minimal turbidity. BChE protein was concentrated on a Pellicon tangential flow filtration system and demonstrated to be highly purified by mass spectrometry. BChE was eluted with 0.1 M tetramethylammonium bromide in 20 mM sodium phosphate pH 8.0. Solvent and detergent reagents used to inactivate lipid enveloped virus, bacteria and protozoa did not bind to Hupresin. The previously utilized anion exchange chromatography step was omitted. The filtered extract was pumped onto a Hupresin affinity column. Butyrylcholinesterase was extracted from frozen Cohn fraction IV-4 in 10 volumes of water pH 6. Our goal was to simplify the purification procedure and increase the yield. Current methods for purifying large quantities of BChE from frozen Cohn fraction IV-4 produce 99% pure enzyme, but the yield is low (21%). Protection from the toxicity of nerve agents is achieved by pretreatment with human butyrylcholinesterase (BChE).
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