Schopfer_2019_PLoS.One_14_e0209795

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Title : Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography - Schopfer_2019_PLoS.One_14_e0209795
Author(s) : Schopfer LM , Lockridge O , David E , Hinrichs SH
Ref : PLoS ONE , 14 :e0209795 , 2019
Abstract : Human butyrylcholinesterase (HuBChE) is being developed as a therapeutic for protection from the toxicity of nerve agents. An enriched source of HuBChE is Cohn fraction IV-4 from pooled human plasma. For the past 40 years, purification of HuBChE has included affinity chromatography on procainamide-Sepharose. The present report supports a new affinity sorbent, Hupresin, for purification of HuBChE from Cohn fraction IV-4. Nine batches of 70-80 kg frozen Cohn fraction were extracted with water, filtered, and chromatographed on 30 L of Q-Ceramic ion exchange sorbent at pH 4.5. The 4% pure Q-eluent was pumped onto 4.2 L Hupresin, where contaminants were washed off with 0.3 M NaCl in 20 mM sodium phosphate pH 8.0, before 99% pure HuBChE was eluted with 0.1 M tetramethylammonium bromide. The average yield was 1.5 g of HuBChE from 80 kg Cohn paste. Recovery of HuBChE was reduced by 90% when the paste was stored at -20 degrees C for 1 year, and reduced 100% when stored at 4 degrees C for 24h. No reduction in HuBChE recovery occurred when paste was stored at -80 degrees C for 3 months or 3 years. Hupresin and procainamide-Sepharose were equally effective at purifying HuBChE from Cohn fraction. HuBChE in Cohn fraction required 1000-fold purification to attain 99% purity, but 15,000-fold purification when the starting material was plasma. HuBChE (P06276) purified from Cohn fraction was a 340 kDa tetramer of 4 identical N-glycated subunits, stable for years in solution or as a lyophilized product.
ESTHER : Schopfer_2019_PLoS.One_14_e0209795
PubMedSearch : Schopfer_2019_PLoS.One_14_e0209795
PubMedID: 30625168

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Schopfer LM, Lockridge O, David E, Hinrichs SH (2019)
Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography
PLoS ONE 14 :e0209795

Schopfer LM, Lockridge O, David E, Hinrichs SH (2019)
PLoS ONE 14 :e0209795