Hupresin Retains Binding Capacity for Butyrylcholinesterase and Acetylcholinesterase after Sanitation with Sodium Hydroxide
Open Access
- 10 October 2017
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Pharmacology
- Vol. 8, 713
- https://doi.org/10.3389/fphar.2017.00713
Abstract
Hupresin is a new affinity resin that binds butyrylcholinesterase (BChE) in human plasma and acetylcholinesterase (AChE) solubilized from red blood cells (RBC). Hupresin is available from the CHEMFORASE company. BChE in human plasma binds to Hupresin and is released with 0.1 M trimethylammonium bromide (TMA) with full activity and 10–15% purity. BChE immunopurified from plasma by binding to immobilized monoclonal beads has fewer contaminating proteins than the one-step Hupresin-purified BChE. However, when affinity chromatography on Hupresin follows ion exchange chromatography at pH 4.5, BChE is 99% pure. The membrane bound AChE, solubilized from human RBC with 0.6% Triton X-100, binds to Hupresin and remains bound during washing with sodium chloride. Human AChE is not released in significant quantities with non-denaturing solvents, but is recovered in 1% trifluoroacetic acid. The denatured, partially purified AChE is useful for detecting exposure to nerve agents by mass spectrometry. Our goal was to determine whether Hupresin retains binding capacity for BChE and AChE after Hupresin is washed with 0.1 M NaOH. A 2 mL column of Hupresin equilibrated in 20 mM TrisCl pH 7.5 was used in seven consecutive trials to measure binding and recovery of BChE from 100 mL human plasma. Between each trial the Hupresin was washed with 10 column volumes of 0.1 M sodium hydroxide. A similar trial was conducted with red blood cell AChE in 0.6% Triton X-100. It was found that the binding capacity for BChE and AChE was unaffected by washing Hupresin with 0.1 M sodium hydroxide. Hupresin could be washed with sodium hydroxide at least seven times without losing binding capacity.Funding Information
- National Institutes of Health (P30CA036727)
This publication has 28 references indexed in Scilit:
- New Huprine Derivatives Functionalized at Position 9 as Highly Potent Acetylcholinesterase InhibitorsChemMedChem, 2011
- Efficacy and physiological effects of human butyrylcholinesterase as a post-exposure therapy against percutaneous poisoning by VX in the guinea-pigChemico-Biological Interactions, 2010
- Immunomagnetic Separation and Quantification of Butyrylcholinesterase Nerve Agent Adducts in Human SerumAnalytical Chemistry, 2010
- A medical health report on individuals with silent butyrylcholinesterase in the Vysya community of IndiaClinica Chimica Acta; International Journal of Clinical Chemistry, 2007
- Albumin, a New Biomarker of Organophosphorus Toxicant Exposure, Identified by Mass SpectrometryToxicological Sciences, 2004
- The Stoichiometry of Protection against Soman and VX Toxicity in Monkeys Pretreated with Human ButyrylcholinesteraseToxicology and Applied Pharmacology, 1997
- A naturally occurring molecular form of human plasma cholinesterase is an albumin conjugateBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1989
- Butyrylcholinesterase in Human Brain and Acetylcholinesterase in Human Plasma: Trace Enzymes Measured by Two‐Site ImmunoassayJournal of Neurochemistry, 1988
- Methode de prepration de la β2-Macroglobuline du serum humainImmunochemistry, 1965
- A "DIRECT-COLORING" THIOCHOLINE METHOD FOR CHOLINESTERASESJournal of Histochemistry & Cytochemistry, 1964