Preprint Article Version 1 This version is not peer-reviewed

Comparison of Hupresin® to Procainamide-Sepharose for Purification of Butyrylcholinesterase and Acetylcholinesterase

Version 1 : Received: 26 June 2018 / Approved: 27 June 2018 / Online: 27 June 2018 (10:52:04 CEST)

How to cite: Lockridge, O.; David, E.; Schopfer, L.M.; Masson, P.; Brazzolotto, X.; Nachon, F. Comparison of Hupresin® to Procainamide-Sepharose for Purification of Butyrylcholinesterase and Acetylcholinesterase. Preprints 2018, 2018060443 (doi: 10.20944/preprints201806.0443.v1). Lockridge, O.; David, E.; Schopfer, L.M.; Masson, P.; Brazzolotto, X.; Nachon, F. Comparison of Hupresin® to Procainamide-Sepharose for Purification of Butyrylcholinesterase and Acetylcholinesterase. Preprints 2018, 2018060443 (doi: 10.20944/preprints201806.0443.v1).

Abstract

Affinity chromatography on procainamide-Sepharose has been an important step in the purification of butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) since its introduction in 1978. The procainamide affinity gel has limitations. In the present report a new affinity gel called Hupresin® was evaluated for its ability to purify truncated, partly-deglycosylated human butyrylcholinesterase (rHuBChE) expressed in a stably transfected CHO cell line. We present a detailed example of the purification of rHuBChE secreted into 3940 mL of serum-free culture medium. The starting material contained 13,163 units of BChE activity (20.9 mg). rHuBChE was purified to homogeneity in a single step by passage over 82 mL of Hupresin® and elution with 0.1 M tetramethylammonium bromide in 20 mM TrisCl pH 7.5. The fraction with the highest specific activity of 630 units/mg contained 11 mg of BChE. Hupresin® is superior to procainamide-Sepharose for purification of BChE, but is not suitable for purifying native AChE because Hupresin® binds AChE so tightly that AChE is desorbed with denaturing solvents such as 50% acetonitrile or 1% trifluoroacetic acid. Procainamide-Sepharose will continue to be useful for purification of AChE.

Subject Areas

Hupresin®; affinity chromatography; procainamide-Sepharose; BChE; AChE

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