Science News

Purification of recombinant human butyrylcholinesterase on Hupresin®.

A new interesting article has been published in J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Dec 1;1102-1103:109-115. doi: 10.1016/j.jchromb.2018.10.026. Epub 2018 Oct 25. and titled:

Purification of recombinant human butyrylcholinesterase on Hupresin®.

Authors of this article are:

Lockridge O, David E, Schopfer LM, Masson P, Brazzolotto X, Nachon F.

A summary of the article is shown below:

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.

Check out the article’s website on Pubmed for more information:



This article is a good source of information and a good way to become familiar with topics such as:

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

.

Categories: Science News