A method for the prevention of thrombin-induced degradation of recombinant proteins

by Dakshinamurthy, Rajalingam; Kathir, Karuppanan Muthusamy; Ananthamurthy, Koteshwara; Adams, Paul D.; Kumar, Thallapuranam Krishnaswamy Suresh

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A new strategy to prevent degradation of recombinant proteins caused by non-specific cleavage by thrombin is described. We demonstrate that degradation due to non-specific cleavage of recombinant protein mediated by thrombin can be completely prevented by separation of thrombin from the recombinant protein on spin columns packed with heparin-sepharose. This method is generally applicable to all recombinant proteins that require the thrombin for the cleavage of affinity tags for purification. To our knowledge, this is the first report of an efficient and reliable method for the separation of residual thrombin from purified recombinant proteins. (c) 2008 Elsevier Inc. All rights reserved.

Journal
Analytical Biochemistry
Volume
375
Issue
2
Year
2008
Start Page
361
ISBN/ISSN
0003-2697
PMID
18279654
DOI
10.1016/j.ab.2008.01.014