Human butyrylcholinesterase for Research

The Human Butyrylcholinesterase Production Facility is capable of producing human butyrylcholinesterase (BChE) in gram quantities from Cohn IV-4 paste (for research purposes only).  Production uses a modification of the process described by Saxena et.al [1], employing hupresin-Sepharose instead of procainamide-Sepharose as the affinity resin.  Process modifications were designed by Dr. Oksana Lockridge (publication pending).

Description

Quality
BChE produced by this method is 98% pure based on SDS PAGE stained with Coomassie Blue, and is 95% tetrameric based on non-denaturing PAGE stained for activity.  Each batch is tested for endotoxin and is certified to have endotoxin levels less than 5 EU/ml.  Refer to the Certificate of Analysis below for details.

Standard Activity Assay Conditions
BChE activity is assayed at 25°C in 0.1 M potassium phosphate buffer, pH 7.0, containing 1 mM butyrylthiocholine and 0.5 mM 5,5'-dithiobis-(2-nitrobenzoic acid).

Unit Definition
One unit will hydrolyze 1.0 µmole of butyrylthiocholine per minute, under standard activity assay conditions.

Activity
Specific activity of the product is 500 units/mg BChE.

Storage
 Store at 2-8°C

Availability and Price

Human butyrylcholinesterase is available for purchase by qualified research institutions or companies.

Catalogue number:

NP06276

1 milligram vial

Available to ship 5/1/2016

Call or e-mail for quote

For all questions related to this service or to obtain BChE contact:

            Larry Schopfer
            Eppley Institute
            University of Nebraska Medical Center
            Omaha, NE 68198-5950
            phone   1-402-559-6305
            fax        1-402-559-7838
            lmschopf@unmc.edu

We are open to collaborations with academic laboratories as a mechanism for reducing cost.  For discussions on collaboration, please contact Dr. Oksana Lockridge.

Production Facility Directors

Dr. Steven Hinrichs

Dr. Oksana Lockridge
olockrid@unmc.edu

References

1    Saxena, Tipparaju, Luo & Doctor; Pilot-scale production of human serum butyrylcholinesterase suitable for use as a bioscavenger against nerve agent toxicity; Process Biochemistry 45, 1313-1318 (2010)

Certificate of Analysis

Human butyrylcholinesterase in phosphate buffered saline
25 mg in 1 mL per vial, under nitrogen
Lot #: October 2015
Activity   12,500 Units/mL
measured in 0.1 M potassium phosphate pH 7, 1 mM butyrylthiocholine, 25˚C
Filter-sterilized through 0.22 µm membrane
MW 340,000; tetramer of 4 identical subunits
Purified from Cohn fraction IV-4 paste
Storage conditions:  4˚C
Endotoxin:  2 EU/mL

Manufactured by the University of Nebraska Medical Center

Figure 1.  SDS gel stained with Coomassie blue.

Samples were reduced and denatured.  Pure HuBChE has a monomer band at 85 kDa and a dimer band at 170 kDa.

Accession number in the Swiss Protein Database:  P06276

Figure 2.  Nondenaturing gel stained for BChE activity.

The pure HuBChE in lanes 1-6 consists predominantly of tetramers.  BChE in human plasma (lanes 7-10) is 95% tetrameric, but includes minor amounts of monomers, two types of dimers, and slow migrating storage bands.
Steven H. Hinrichs, MD                                                                   Oksana Lockridge, PhD
University of Nebraska Medical Center                                      University of Nebraska Medical Center
Omaha, NE 68198-5900                                                                   Omaha, NE 68198-5950 
shinrich@unmc.edu                                                                           olockrid@unmc.edu