Recombinant human butyrylcholinesterase from milk of transgenic animals to protect against organophosphate poisoning
- Yue-Jin Huang*,†,
- Yue Huang*,
- Hernan Baldassarre*,
- Bin Wang*,‡,
- Anthoula Lazaris*,§,
- Martin Leduc*,¶,
- Annie S. Bilodeau*,
- Annie Bellemare*,
- Mélanie Côté*,
- Peter Herskovits*,
- Madjid Touati*,
- Carl Turcotte*,
- Loredana Valeanu*,
- Nicolas Lemée*,
- Harvey Wilgus*,
- Isabelle Bégin*,
- Bhim Bhatia*,
- Khalid Rao*,
- Nathalie Neveu*,
- Eric Brochu*,
- Janice Pierson*,
- Duncan K. Hockley*,
- Douglas M. Cerasoli‖,
- David E. Lenz‖,
- Costas N. Karatzas*,**, and
- Solomon Langermann*
- *PharmAthene Canada, Inc., 7150 Alexander-Fleming, Montreal, QC, Canada H4S 2C8; and
- ‖United States Army Medical Research Institute of Chemical Defense, 3100 Ricketts Point Road, Aberdeen Proving Ground, MD 21010-5400
-
Edited by R. Michael Roberts, University of Missouri, Columbia, MO, and approved June 22, 2007 (received for review March 23, 2007)
Abstract
Dangerous organophosphorus (OP) compounds have been used as insecticides in agriculture and in chemical warfare. Because exposure to OP could create a danger for humans in the future, butyrylcholinesterase (BChE) has been developed for prophylaxis to these chemicals. Because it is impractical to obtain sufficient quantities of plasma BChE to treat humans exposed to OP agents, the production of recombinant BChE (rBChE) in milk of transgenic animals was investigated. Transgenic mice and goats were generated with human BChE cDNA under control of the goat β-casein promoter. Milk from transgenic animals contained 0.1–5 g/liter of active rBChE. The plasma half-life of PEGylated, goat-derived, purified rBChE in guinea pigs was 7-fold longer than non-PEGylated dimers. The rBChE from transgenic mice was inhibited by nerve agents at a 1:1 molar ratio. Transgenic goats produced active rBChE in milk sufficient for prophylaxis of humans at risk for exposure to OP agents.
Footnotes
- †To whom correspondence should be addressed at: PharmAthene Canada, Inc., 7150 Alexander-Fleming, Montreal, QC, Canada H4S 2C8. Email: yjhuang{at}pharmathene.ca
-
↵ ‡Present address: Advanced Reproductive Care Center, 10301 Hagen Ranch Road, Boynton Beach, FL 33437.
-
↵ §Present address: Quebec Transgenic Research Network, McGill University, 3655 Promenade Sir William Osler, Montreal, QC, Canada H3G 1Y6.
-
↵ ¶Present address: Research Center, Hôpital Ste-Justine, 3175 Cote Ste. Catherine, Montreal, QC, Canada H3T 1C5.
-
↵**Present address: CNKonsulting and Aegis Bioresearch, 251 Sherwood Road, Beaconsfield, QC, Canada H9W 2H4.
-
Author contributions: Y.-J.H., H.B., A.L., and C.N.K. designed research; Y.-J.H., Y.H., H.B., B.W., A.L., M.L., A.S.B., A.B., M.C., P.H., M.T., C.T., L.V., N.L., H.W., I.B., B.B., K.R., N.N., E.B., J.P., D.K.H., D.M.C., D.E.L., and C.N.K. performed research; Y.-J.H., Y.H., H.B., B.W., A.L., H.W., D.K.H., D.M.C., D.E.L., C.N.K., and S.L. analyzed data; and Y.-J.H., A.L., C.N.K., and S.L. wrote the paper.
-
The authors declare no conflict of interest.
-
This article is a PNAS Direct Submission.
-
This article contains supporting information online at www.pnas.org/cgi/content/full/0702756104/DC1.
- Abbreviations:
- BChE,
- butyrylcholinesterase;
- FVB,
- friend virus B-type;
- huBChE,
- human butyrylcholinesterase;
- OP,
- organophosphorus compounds;
- rBChE,
- recombinant butyrylcholinesterase;
- SEC,
- size exclusion chromatography.
-
Freely available online through the PNAS open access option.
- © 2007 by The National Academy of Sciences of the USA
