Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies

T.Z. Yuan; Callum Ormonde; S.T. Kudlacek; S. Kunche; J.N. Smith; W.A. Brown; K.M. Pugliese; T.J. Olsen; M. Iftikhar; C.L. Raston; G.A. Weiss

doi:10.1002/cbic.201402427

Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies

T.Z. Yuan, Callum Ormonde, S.T. Kudlacek, S. Kunche, J.N. Smith, W.A. Brown, K.M. Pugliese, T.J. Olsen, M. Iftikhar, C.L. Raston, G.A. Weiss

School of Molecular Sciences

Research output: Contribution to journal › Article › peer-review

80 Citations (Scopus)

Abstract

© 2015 Wiley-VCH Verlag GmbH & Co. KGaA. 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 language	English
Pages (from-to)	393-396
Journal	ChemBioChem
Volume	16
Issue number	3
DOIs	https://doi.org/10.1002/cbic.201402427
Publication status	Published - 2015

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title = "Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies",

abstract = "{\textcopyright} 2015 Wiley-VCH Verlag GmbH & Co. KGaA. 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.",

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T1 - Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies

AU - Yuan, T.Z.

AU - Ormonde, Callum

AU - Kudlacek, S.T.

AU - Kunche, S.

AU - Smith, J.N.

AU - Brown, W.A.

AU - Pugliese, K.M.

AU - Olsen, T.J.

AU - Iftikhar, M.

AU - Raston, C.L.

AU - Weiss, G.A.

PY - 2015

Y1 - 2015

N2 - © 2015 Wiley-VCH Verlag GmbH & Co. KGaA. 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.

AB - © 2015 Wiley-VCH Verlag GmbH & Co. KGaA. 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.

U2 - 10.1002/cbic.201402427

DO - 10.1002/cbic.201402427

M3 - Article

SN - 1439-4227

VL - 16

SP - 393

EP - 396

JO - ChemBioChem

JF - ChemBioChem

IS - 3

ER -

Shear-stress-mediated refolding of proteins from aggregates and inclusion bodies

Abstract

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