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

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.