A highly stable minimally processed plant-derived recombinant acetylcholinesterase for nerve agent detection in adverse conditions.
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.
PlantVax Inc, Rockville, MD, 20850.
Yvonne J
YJ
Rosenberg
FLIR Systems Inc, Pittsburgh, PA, 15238.
Jeremy
J
Walker
PlantVax Inc, Rockville, MD, 20850.
Xiaoming
X
Jiang
FLIR Systems Inc, Pittsburgh, PA, 15238.
Scott
S
Donahue
FLIR Systems Inc, Pittsburgh, PA, 15238.
Jason
J
Robosky
Institute for molecular Biotechnology, RWTH Aachen University, Aachen, Germany.
Markus
M
Sack
PlantVax Inc, Rockville, MD, 20850.
Jonathan
J
Lees
PlantVax Inc, Rockville, MD, 20850.
Lori
L
Urban
D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D002800:Cholinesterase Inhibitors / Q000032:analysis / Q000737:chemistry*; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D057230:Limit of Detection; D000067397:Nerve Agents / Q000032:analysis / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D030821:Plants, Genetically Modified / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D014026:Nicotiana / Q000235:genetics
D002800:Cholinesterase Inhibitors; D000067397:Nerve Agents; D011994:Recombinant Proteins; D000110:Acetylcholinesterase