Literature DB >> 18279654

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

Dakshinamurthy Rajalingam1, Karuppanan Muthusamy Kathir, Koteshwara Ananthamurthy, Paul D Adams, Thallapuranam Krishnaswamy Suresh Kumar.   

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.

Entities:  

Mesh:

Substances:

Year:  2008        PMID: 18279654      PMCID: PMC2614318          DOI: 10.1016/j.ab.2008.01.014

Source DB:  PubMed          Journal:  Anal Biochem        ISSN: 0003-2697            Impact factor:   3.365


  8 in total

Review 1.  Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems.

Authors:  K Terpe
Journal:  Appl Microbiol Biotechnol       Date:  2002-11-07       Impact factor: 4.813

Review 2.  Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems.

Authors:  Kay Terpe
Journal:  Appl Microbiol Biotechnol       Date:  2006-06-22       Impact factor: 4.813

Review 3.  Recombinant protein expression in Escherichia coli.

Authors:  F Baneyx
Journal:  Curr Opin Biotechnol       Date:  1999-10       Impact factor: 9.740

Review 4.  Determinants of thrombin specificity.

Authors:  E Di Cera; A M Cantwell
Journal:  Ann N Y Acad Sci       Date:  2001       Impact factor: 5.691

5.  Comparison of affinity tags for protein purification.

Authors:  Jordan J Lichty; Joshua L Malecki; Heather D Agnew; Daniel J Michelson-Horowitz; Song Tan
Journal:  Protein Expr Purif       Date:  2005-05       Impact factor: 1.650

6.  Quantitative characterization of the thrombin-heparin interaction. Discrimination between specific and nonspecific binding models.

Authors:  S T Olson; H R Halvorson; I Björk
Journal:  J Biol Chem       Date:  1991-04-05       Impact factor: 5.157

7.  Copper binding affinity of S100A13, a key component of the FGF-1 nonclassical copper-dependent release complex.

Authors:  Vaithiyalingam Sivaraja; Thallapuranam Krishnaswamy Suresh Kumar; Dakshinamurthy Rajalingam; Irene Graziani; Igor Prudovsky; Chin Yu
Journal:  Biophys J       Date:  2006-06-09       Impact factor: 4.033

Review 8.  Strategies for achieving high-level expression of genes in Escherichia coli.

Authors:  S C Makrides
Journal:  Microbiol Rev       Date:  1996-09
  8 in total
  2 in total

1.  Simple and Efficient Purification of Recombinant Proteins Using the Heparin-Binding Affinity Tag.

Authors:  Srinivas Jayanthi; Ravi Kumar Gundampati; Thallapuranam Krishnaswamy Suresh Kumar
Journal:  Curr Protoc Protein Sci       Date:  2017-11-01

2.  Transgene-host cell interactions mediate significant influences on the production, stability, and function of recombinant canine FVIII.

Authors:  Bredon Crawford; Margareth C Ozelo; Kenichi Ogiwara; James Ahlin; Silvia Albanez; Carol Hegadorn; Lori Harpell; Christine Hough; David Lillicrap
Journal:  Mol Ther Methods Clin Dev       Date:  2015-11-18       Impact factor: 6.698

  2 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.