Purification of recombinant human butyrylcholinesterase on Hupresin®

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

Research output: Contribution to journalArticle

2 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)109-115
Number of pages7
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume1102-1103
DOIs
StatePublished - Dec 1 2018

Keywords

  • Acetylcholinesterase
  • Affinity chromatography
  • Butyrylcholinesterase
  • Hupresin®
  • Procainamide-Sepharose

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Purification of recombinant human butyrylcholinesterase on Hupresin®'. Together they form a unique fingerprint.

  • Cite this