Rosenberg Yvonne J, Walker Jeremy, Jiang Xiaoming, Donahue Scott, Robosky Jason, Sack Markus, Lees Jonathan, Urban Lori
PlantVax Inc, Rockville, MD, 20850.
FLIR Systems Inc, Pittsburgh, PA, 15238.
Sci Rep. 2015 Aug 13;5:13247. doi: 10.1038/srep13247.
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.
尽管最近瞬态植物系统的创新已能够在1-2周内生产克级数量的蛋白质,但由于技术挑战和高昂的下游加工成本,很少有成果转化为实际应用。在此,我们报告了使用本氏烟草/p19系统高水平生产一种工程重组人乙酰胆碱酯酶(rAChE),该酶在经过最少加工的叶提取物中高度稳定。冻干的澄清提取物在70°C下可长期保存,复溶后可用于多种设备,以检测0°C至50°C表面的有机磷酸酯(OP)神经毒剂和农药。最近叙利亚使用沙林毒气事件凸显了对神经毒剂检测以及防范和应急反应所需对策的迫切需求。绕过繁琐且昂贵的下游工艺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。