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Sugaring out: A new method for removal of acetonitrile from preparative RP-HPLC eluent for protein purification

Paper ID Volume ID Publish Year Pages File Format Full-Text
35511 45093 2010 5 PDF Available
Title
Sugaring out: A new method for removal of acetonitrile from preparative RP-HPLC eluent for protein purification
Abstract

Reverse phase-high pressure liquid chromatography (RP-HPLC) with an acetonitrile–water mixture as the eluent is widely used for purification of proteins. The separation of acetonitrile (ACN) in RP-HPLC eluent is important for protein recovery. Cooling below subzero temperature and salting out have been used to remove ACN, each with its limitations. In this work we have explored the use of sugaring-out, a new phase separation method developed at University of Illinois for the separation of ACN from a simulated preparative RP-HPLC effluent. The effect of glucose concentration, temperature, and initial amount of ACN in the effluent on phase separation was investigated. Results showed that a good phase separation can be achieved at near room temperature (18 °C). With the optimized conditions, we found that more than 60% (w/w) of ACN was removed and more than 95% (w/w) of water-soluble proteins (bovine serum albumin, trypsin, and pepsin) were recovered.

Keywords
Liquid–liquid extraction; Solvent extraction; Sugaring-out; Proteins; Acetonitrile
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Sugaring out: A new method for removal of acetonitrile from preparative RP-HPLC eluent for protein purification
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 45, Issue 10, October 2010, Pages 1672–1676
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us