Overview of Approaches to Preventing and Avoiding Proteolysis During Expression and Purification of Proteins

Barry J. Ryan1, Gary T. Henehan1

1 Food Science and Environmental Health, Dublin Institute of Technology, Dublin, Ireland
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 5.25
DOI:  10.1002/0471140864.ps0525s71
Online Posting Date:  February, 2013
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Proteases are enzymes that cleave proteins. They occur widely in nature and play a fundamental role in cellular homeostasis; however, their presence can result in unwanted protein degradation during recombinant protein expression and purification. This unit introduces proteases, specifically focusing on the types commonly encountered during production of recombinant proteins. The strategies used to avoid and to prevent proteolysis are also described, with extensive consideration of the molecular, technical, and logistic methodologies involved. Curr. Protoc. Protein Sci. 71:5.25.1‐5.25.7. © 2013 by John Wiley & Sons, Inc.

Keywords: protease; proteolysis; recombinant protein

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Table of Contents

  • Introduction
  • Strategies for Avoiding Proteolysis
  • Strategies for Preventing Proteolysis
  • Special Problem of Expression of Protein Fragments
  • Summary
  • Literature Cited
  • Tables
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Literature Cited

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