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Protein Chromatography pp 53–69Cite as

Avoiding Proteolysis During Protein Purification

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1485))

Abstract

All cells contain proteases which hydrolyze the peptide bonds between amino acids in 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; firstly inhibition of proteolysis in situ, followed by the early separation of the protease from the protein of interest via chromatographical 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. Here, commonly used approaches to reducing or avoiding proteolysis during protein purification and subsequent chromatography are reviewed.

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Author information

Authors and Affiliations

  1. School of Food Science and Environmental Health, Dublin Institute of Technology, Cathal Brugha Street, Dublin 1, Republic of Ireland

    Barry J. Ryan & Gary T. Henehan

Authors
  1. Barry J. Ryan
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  2. Gary T. Henehan
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Correspondence to Barry J. Ryan .

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Editors and Affiliations

  1. School of Biotechnology, Dublin City University, Dublin, Ireland

    Dermot Walls

  2. Department of Applied Sciences, Dundalk Institute of Technology, Dublin, Ireland

    Sinéad T. Loughran

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Ryan, B.J., Henehan, G.T. (2017). Avoiding Proteolysis During Protein Purification. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 1485. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6412-3_4

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  • DOI: https://doi.org/10.1007/978-1-4939-6412-3_4

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  • Publisher Name: Humana Press, New York, NY

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