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

Authors

Dakshinamurthy Rajalingam, University of Arkansas, Fayetteville
Karuppanan Muthusamy Kathir, University of Arkansas, Fayetteville
Koteshwara Ananthamurthy, University of Arkansas, Fayetteville
Paul D. Adams, University of Arkansas, FayettevilleFollow
Thallapuranam Krishnaswamy Suresh Kumar, University of Arkansas, FayettevilleFollow

Document Type

Article - Abstract Only

Publication Date

4-15-2008

Keywords

Recombinant proteins, thrombin, affinity tags, purification

Abstract

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.

Comments

Principal Investigator: Paul Adams

Acknowledgements: The work was supported by grants from the National Science Foundation (NSF 0726004) to P.D.A. The National Institute of Health (NIH NCRR COBRE Grant 1 P20RR15569), the Arkansas Bioscience Institute to T.K.S.K.

Citation

Rajalingam, D., Kathir, K. M., Ananthamurthy, K., Adams, P. D., & Kumar, T. K. (2008). A method for the prevention of thrombin-induced degradation of recombinant proteins. Analytical Biochemistry, 375 (2), 361-363. https://doi.org/10.1016/j.ab.2008.01.014