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Protein Extraction and Purification by Differential Solubilization

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2699))

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Abstract

The preparation of purified soluble proteins for biochemical studies is essential and the solubility of a protein of interest in various media is central to this process. Selectively altering the solubility of a protein is a rapid and economical step in protein purification and is based on exploiting the inherent physicochemical properties of a polypeptide. Precipitation of proteins, released from cells upon lysis, is often used to concentrate a protein of interest before further purification steps (e.g., ion exchange chromatography, size exclusion chromatography etc).

Recombinant proteins may be expressed in host cells as insoluble inclusion bodies due to various influences during overexpression. Such inclusion bodies can often be solubilized to be reconstituted as functional, correctly folded proteins.

In this chapter, we examine strategies for extraction/precipitation/solubilization of proteins for protein purification. We also present bioinformatic tools to aid in understanding a protein’s propensity to aggregate/solubilize that will be a useful starting point for the development of protein extraction, precipitation, and selective re-solubilization procedures.

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Author information

Authors and Affiliations

  1. School of Food Science and Environmental Health, Technological University Dublin, Grangegorman, Dublin, Ireland

    Barry J. Ryan, Gemma K. Kinsella & Gary T. Henehan

Authors
  1. Barry J. Ryan
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  2. Gemma K. Kinsella
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  3. Gary T. Henehan
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Corresponding author

Correspondence to Gary T. Henehan .

Editor information

Editors and Affiliations

  1. Department of Life and Health Sciences, Dundalk Institute of Technology, Dundalk, Louth, Ireland

    Sinéad T Loughran

  2. National Institute for Bioprocessing Research and Training (NIBRT), Blackrock, Dublin, Ireland

    John Joseph Milne

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ryan, B.J., Kinsella, G.K., Henehan, G.T. (2023). Protein Extraction and Purification by Differential Solubilization. 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_17

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  • DOI: https://doi.org/10.1007/978-1-0716-3362-5_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3361-8

  • Online ISBN: 978-1-0716-3362-5

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