title: In vitro and in vivo characterization of recombinant human butyrylcholinesterase (Protexia) as a potential nerve agent bioscavenger.
abstract: Previous studies in rodents and nonhuman primates have demonstrated that pretreatment with cholinesterases can provide significant protection against behavioral and lethal effects of nerve agent intoxication. Human butyrylcholinesterase (HuBuChE) purified from plasma has been shown to protect against up to 5 x LD50s of nerve agents in guinea pigs and non-human primates, and is currently being explored as a bioscavenger pretreatment for human use. A recombinant form of HuBuChE has been expressed in the milk of transgenic goats as a product called Protexia. Protexia was supplied by Nexia Biotechnologies (Que., Canada) as a purified solution with a specific activity of 600 U/mg. Initial in vitro studies using radiolabeled 3H-soman or 3H-DFP (diisopropyl fluorophosphate) demonstrated that these inhibitors specifically bind to Protexia. When Protexia was mixed with soman, sarin, tabun or VX using varying molar ratios of enzyme to nerve agent (8:1, 4:1, 1:1 and 1:4, respectively), the data indicated that 50% inhibition of enzyme activity occurs around the 1:1 molar ratio for each of the nerve agents. Protexia was further characterized for its interaction with pyridostigmine bromide and six unique carbamate inhibitors of cholinesterase. IC50 and Ki values for Protexia were determined to be very similar to those of HuBuChE purified from human plasma. These data suggest that Protexia has biochemical properties very similar to those HuBuChE when compared in vitro. Together these data the continued development of the goat milk-derived recombinant HuBuChE Protexia as a potential bioscavenger of organophosphorus nerve agents.
authors.affiliation: Pharmacology Division, USAMRICD, Aberdeen Proving Ground, MD 21010, USA.
authors.forename: D M
authors.initials: DM
authors.lastname: Cerasoli
authors.forename: E M
authors.initials: EM
authors.lastname: Griffiths
authors.forename: B P
authors.initials: BP
authors.lastname: Doctor
authors.forename: A
authors.initials: A
authors.lastname: Saxena
authors.forename: J M
authors.initials: JM
authors.lastname: Fedorko
authors.forename: N H
authors.initials: NH
authors.lastname: Greig
authors.forename: Q S
authors.initials: QS
authors.lastname: Yu
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.forename: H
authors.initials: H
authors.lastname: Wilgus
authors.forename: C N
authors.initials: CN
authors.lastname: Karatzas
authors.forename: I
authors.initials: I
authors.lastname: Koplovitz
authors.forename: D E
authors.initials: DE
authors.lastname: Lenz
mesh_terms: D000818:Animals; D002091:Butyrylcholinesterase / Q000737:chemistry / Q000494:pharmacology*; D002219:Carbamates / Q000037:antagonists & inhibitors; D006041:Goats; D006801:Humans; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D009498:Neurotoxins / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology
chemical_list: D002219:Carbamates; D009498:Neurotoxins; D011994:Recombinant Proteins; D002091:Butyrylcholinesterase