A highly stable minimally processed plant-derived recombinant acetylcholinesterase for nerve agent detection in adverse conditions.
A highly stable minimally processed plant-derived recombinant acetylcholinesterase for nerve agent detection in adverse conditions.
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DOI：
10.1038/srep13247
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发表时间：
2015-08-13
影响因子：
4.6
通讯作者：
Urban L
中科院分区：
文献类型：
--
作者：
Rosenberg YJ;Walker J;Jiang X;Donahue S;Robosky J;Sack M;Lees J;Urban L
尽管瞬时植物系统的最新创新已在 1-2 周内实现了克级蛋白质，但由于技术挑战和高昂的下游加工成本，很少有转化为应用。在这里，我们报告了使用本塞姆氏烟草/p19系统高水平生产了一种工程重组人乙酰胆碱酯酶（rAChE），该酶在经过最低限度加工的叶提取物中高度稳定。冻干澄清提取物可在 70°C 下长期保存，并且在复溶后可用于多种设备，以检测 0°C 至 50°C 表面上的有机磷 (OP) 神经毒剂和农药。最近在叙利亚使用沙林事件凸显了备灾和应急响应所需的神经毒剂检测和对策的迫切需要。绕过繁琐且昂贵的下游流程，使我们能够充分利用瞬时生产系统的速度、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。
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.