Document Type
Book Chapter
Disciplines
1. NATURAL SCIENCES
Abstract
All cells contain proteases, which hydrolyze the peptide bonds between amino acids of a protein backbone. Typically, proteases are prevented from nonspecific proteolysis by regulation and by their physical separation into different subcellular compartments; however, this segregation is not retained during cell lysis, which is the initial step in any protein isolation procedure. Prevention of proteolysis during protein purification often takes the form of a two-pronged approach: first, inhibition of proteolysis in situ, followed by the early separation of the protease from the protein of interest via chromatographic purification. Protease inhibitors are routinely used to limit the effect of the proteases before they are physically separated from the protein of interest via column chromatography. In this chapter, commonly used approaches to reducing or avoiding proteolysis during protein expression and purification are reviewed.
DOI
https://doi.org/10.1007/978-1-0716-3362-5_6
Recommended Citation
Henehan, Gary T.; Ryan, Barry J.; and Kinsella, Gemma K., "Approaches to Avoid Proteolysis During Protein Expression and Purification" (2023). Books/Book Chapters/ Proceedings. 22.
https://arrow.tudublin.ie/schfsehbk/22
Funder
This research received no external funding
Creative Commons License
This work is licensed under a Creative Commons Attribution-Share Alike 4.0 International License.
Publication Details
https://link.springer.com/protocol/10.1007/978-1-0716-3362-5_6
Henehan, G.T., Ryan, B.J., Kinsella, G.K. (2023). Approaches to Avoid Proteolysis During Protein Expression and Purification. In: Loughran, S.T., Milne, J.J. (eds) Protein Chromatography. Methods in Molecular Biology, vol 2699. Humana, New York, NY.
https://doi.org/10.1007/978-1-0716-3362-5_6