期刊
PROCESS BIOCHEMISTRY
卷 45, 期 8, 页码 1313-1318出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2010.04.021
关键词
Human serum butyrylcholinesterase; Cohn fraction IV-4; Organophosphorus nerve agents; Bioscavenger; Pilot-scale production
Human serum butyrylcholinesterase (Hu BChE) is currently the most appropriate candidate for the prophylaxis of humans against organophosphate (OP) nerve agent toxicity. It is estimated that a dose of 200 mg will protect a human against 2 x LD(50) of soman, which means that gram quantities of enzyme are needed for human clinical studies. Toward this effort, we report the development of the first procedure that is suitable for the pilot-scale purification of Hu BChE from Cohn fraction IV-4 paste. This procedure involved resuspension of Cohn fraction IV-4 paste, followed by procainamide affinity and DEAE anion-exchange chromatography. The procedure yielded 6-7 g (4.3-5 million U) of purified enzyme from 80 kg of Cohn fraction IV-4 paste. The enzyme was >97% pure as judged by a specific activity of similar to 700 U/mg and a major band with a subunit molecular weight of 85 kDa on SDS-PAGE. The high yield and purity obtained suggest that this manufacturing procedure is suitable for the pre-clinical production of Hu BChE. Published by Elsevier Ltd.
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