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Purification of Recombinant Human Butyrylcholinesterase on Hupresin®

Purification of Recombinant Human Butyrylcholinesterase on Hupresin®

Oksana Lockridge, Emilie David, Lawrence M Schopfer, Patrick Masson, Xavier Brazzolotto, Florian Nachon

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Dec 1;1102-1103:109-115.

PMID: 30384187

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, recombinant human butyrylcholinesterase (rHuBChE) expressed in a stably transfected Chinese Hamster Ovary 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® eluted 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 not released with buffers, but is desorbed with denaturing solvents such as 50% acetonitrile or 1% trifluoroacetic acid. Procainamide-Sepharose will continue to be useful for purification of AChE.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP64200	Tetramethylammonium bromide		64-20-0	Price

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