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A highly stable minimally processed plant-derived recombinant acetylcholinesterase for nerve agent detection in adverse conditions

机译:一种高度稳定的经过最少加工的植物来源的重组乙酰胆碱酯酶用于在不利条件下检测神经毒剂

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摘要

Although recent innovations in transient plant systems have enabled gram quantities of proteins in 1–2 weeks, very few have been translated into applications due to technical challenges and high downstream processing costs. Here we report high-level production, using a Nicotiana benthamiana/p19 system, of an engineered recombinant human acetylcholinesterase (rAChE) that is highly stable in a minimally processed leaf extract. Lyophylized clarified extracts withstand prolonged storage at 70 °C and, upon reconstitution, can be used in several devices to detect organophosphate (OP) nerve agents and pesticides on surfaces ranging from 0 °C to 50 °C. The recent use of sarin in Syria highlights the urgent need for nerve agent detection and countermeasures necessary for preparedness and emergency responses. Bypassing cumbersome and expensive downstream processes has enabled us to fully exploit the speed, low cost and scalability of transient production systems resulting in the first successful implementation of plant-produced rAChE into a commercial biotechnology product.
机译:尽管近期在过渡性植物系统中的创新已使1-2周内达到克级的蛋白质水平,但由于技术挑战和较高的下游加工成本,很少将其转化为应用。在这里,我们报告了使用本特烟草(Nicotiana benthamiana)/ p19系统高水平生产的工程重组人乙酰胆碱酯酶(rAChE),该酶在最少加工的叶提取物中高度稳定。冻干的澄清提取物可在70°C的温度下长期保存,并且经过重构,可用于多种设备中,以检测0°C至50°C范围内的有机磷酸盐(OP)神经毒剂和农药。叙利亚最近对沙林蛋白的使用突显了对神经毒剂检测的迫切需求,以及为备灾和应急响应所必需的对策。绕开繁琐而昂贵的下游过程,使我们能够充分利用瞬态生产系统的速度,低成本和可扩展性,从而首次成功地将植物生产的rAChE应用于商业生物技术产品。

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