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Differential Precipitation and Solubilization of Proteins

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Protein Chromatography

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

Abstract

Differential protein precipitation is a rapid and economical step in protein purification and is based on exploiting the inherent physicochemical properties of the polypeptide. Precipitation of recombinant proteins, lysed from the host cell, is commonly used to concentrate the protein of choice before further polishing steps with more selective purification columns (e.g., His-Tag, Size Exclusion, etc.). Recombinant proteins can also precipitate naturally as inclusion bodies due to various influences during overexpression in the host cell. Although this phenomenon permits easier initial separation from native proteins, these inclusion bodies must carefully be differentially solubilized so as to reform functional, correctly folded proteins. Here, appropriate bioinformatics tools to aid in understanding a protein’s propensity to aggregate and solubilize are explored as a backdrop for a typical protein extraction, precipitation, and selective resolubilization procedure, based on a recombinantly expressed protein.

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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 & Gemma K. Kinsella

Authors
  1. Barry J. Ryan
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  2. Gemma K. Kinsella
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Corresponding author

Correspondence to Barry J. Ryan .

Editor information

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., Kinsella, G.K. (2017). Differential Precipitation and Solubilization of Proteins. 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_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6412-3_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6410-9

  • Online ISBN: 978-1-4939-6412-3

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