Large-scale purification and long-term stability of human butyrylcholinesterase: a potential bioscavenger drug

Journal of Biochemical and Biophysical Methods
J GrunwaldY Ashani

Abstract

Butyrylcholinesterase from human plasma (HuBChE) is a potential drug candidate for detoxification of certain harmful chemicals that contain carboxylic or phosphoric acid ester bonds. Large quantities of purified HuBChE, displaying a high stability upon long-term storage, are required for the evaluation of its therapeutic capacity and its pharmaceutical properties. Several modifications of a previously reported procedure enabled us to purify the enzyme > 15,000-fold from pools of up to 100 1 of human plasma. The three-step procedure is based on precipitation of plasma proteins by ammonium sulfate (step I) and batch adsorption of HuBChE on procainamide-Sepharose 4B gel (step II). Ammonium sulfate was also employed in the third stage to fractionate the final product from procainamide-containing HuBChE solution. The overall yield (63%) of electrophoretically pure enzyme was significantly higher than that previously reported (34%) for the purification of HuBChE from 12.5 1 of plasma or from 5 kg of Cohn fraction IV-4. Purified HuBChE was stored at 5 degrees C in 10 mM phosphate buffer (pH 7.4) containing 1 mM EDTA and 0.02% NaN3. The specific activity, protein migration on gel electrophoresis, thermostability at 54 degrees C and the...Continue Reading

References

May 1, 1977·Life Sciences·D J StewartW Kalow
Nov 1, 1979·Analytical Biochemistry·E G DelMarM C Geokas
Jan 1, 1976·Methods in Enzymology·J Porath, R Axén
Jun 1, 1968·Journal of Pharmaceutical Sciences·S T Christian, J G Beasley
Apr 1, 1983·The Biochemical Journal·J S RalstonA L Chasson
Jan 1, 1981·Advances in Clinical Chemistry·S S BrownC L Woronick
Nov 1, 1994·Toxicon : Official Journal of the International Society on Toxinology·O JbiloO Lockridge
Dec 1, 1993·Pharmacology, Biochemistry, and Behavior·R BrandeisY Ashani
Jul 1, 1961·Biochemical Pharmacology·G L ELLMANR M FEATHER-STONE
Mar 1, 1964·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·M J KARNOVSKY, L ROOTS

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Citations

May 4, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Tama EvronTsafrir S Mor
Dec 25, 2008·BMC Biotechnology·Ryan R WoodsTsafrir S Mor
Jul 5, 2011·Toxicology and Applied Pharmacology·Mariya LiyasovaOksana Lockridge
Jul 8, 2008·Protein Expression and Purification·Ashima SaxenaBhupendra P Doctor
Dec 21, 2006·Neurochemistry International·Luis F Sánchez del CampoFrancisco J Campoy
Oct 22, 2008·Journal of Applied Toxicology : JAT·He LiOksana Lockridge
Apr 1, 2010·Plant Biotechnology Journal·Brian C GeyerTsafrir S Mor
Jul 13, 2004·Experimental Neurology·Sultan DarveshJ Andrew Armour
Nov 5, 2005·Chemico-biological Interactions·Brian C GeyerTsafrir S Mor
Oct 7, 2004·Journal of Toxicology. Clinical Toxicology·Muhammet GüvenFevzi Altuntaş
Jun 12, 2014·The Biochemical Journal·Marilynn A LarsonSteven H Hinrichs
Oct 27, 2012·Chemico-biological Interactions·Katarina MrvovaAnna Hrabovska
Jul 10, 2009·Clinical Toxicology : the Official Journal of the American Academy of Clinical Toxicology and European Association of Poisons Centres and Clinical Toxicologists·Lucie MusilovaDaniel Jun
Aug 18, 2004·Human & Experimental Toxicology·Muhammet GüvenBülent Eser
Nov 6, 2003·Toxicological Sciences : an Official Journal of the Society of Toxicology·Yacov Ashani, Shlomi Pistinner
Jun 22, 2006·Pharmacogenetics and Genomics·Indumathi ManoharanRathnam Boopathy
Nov 2, 2018·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Oksana LockridgeFlorian Nachon
Sep 12, 2017·Chemical Research in Toxicology·Alicia J DaffernerOksana Lockridge
Dec 17, 2009·Archives of Biochemistry and Biophysics·Patrick Masson, Oksana Lockridge

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