title: Purification of transgenic plant-derived recombinant human acetylcholinesterase-R.
abstract: Nicotiana benthamiana plants were engineered to express a codon-optimized gene encoding the human acetylcholinesterase-R (AChE) isoform. The transgenic plants expressed the protein at >0.4% of total soluble protein, and the plant-produced enzyme was purified to homogeneity. Following lysis, procainamide affinity chromatography and anion-exchange chromatography, more than 400-fold purification was achieved and electrophoretic purity was obtained. This pure protein is kinetically indistinguishable from the only commercially available source of human acetylcholinesterase, which is produced in mammalian cell culture. Thus, we have demonstrated a model system for the production of acetylcholinesterase, which is not susceptible to the quantitative limitations or mammalian pathogens associated with purification from mammalian cell culture or human serum.
authors.affiliation: School of Life Sciences and The Biodesign Institute, P.O. Box 874501, Arizona State University, Tempe, AZ 85287-4501, USA.
authors.forename: Brian C
authors.initials: BC
authors.lastname: Geyer
authors.forename: Mrinalini
authors.initials: M
authors.lastname: Muralidharan
authors.forename: Irene
authors.initials: I
authors.lastname: Cherni
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Doran
authors.forename: Samuel P
authors.initials: SP
authors.lastname: Fletcher
authors.forename: Tama
authors.initials: T
authors.lastname: Evron
authors.forename: Hermona
authors.initials: H
authors.lastname: Soreq
authors.forename: Tsafrir S
authors.initials: TS
authors.lastname: Mor
mesh_terms: D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics*; D014027:Tobacco Mosaic Virus / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D000110:Acetylcholinesterase