Shear-Stress-Mediated Refolding of Proteins from Aggregates and Inclusion Bodies

Tom Yuan, Callum Ormonde, Stephan Kudlacek, Sameeran Kunche, Joshua Smith, William Brown, Kaitlin Pugliese, Tivoli Olsen, Mariam Ifikhar, Colin Raston, Gregory Weiss

    Research output: Contribution to journalArticlepeer-review

    55 Citations (Scopus)

    Abstract

    Recombinant protein overexpression of large proteins in bacteria often results in insoluble and misfolded proteins directed to inclusion bodies. We report the application of shear stress in micrometer-wide, thin fluid films to refold boiled hen egg white lysozyme, recombinant hen egg white lysozyme, and recombinant caveolin-1. Furthermore, the approach allowed refolding of a much larger protein, cAMP-dependent protein kinase A (PKA). The reported methods require only minutes, which is more than 100 times faster than conventional overnight dialysis. This rapid refolding technique could significantly shorten times, lower costs, and reduce waste streams associated with protein expression for a wide range of industrial and research applications.

    Original languageEnglish
    Pages (from-to)393-396
    Number of pages4
    JournalChembiochem
    Volume16
    Issue number3
    DOIs
    Publication statusPublished - 9 Feb 2015

    Keywords

    • Green chemistry
    • Protein expression
    • Protein folding
    • Shear stress

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