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Transient Expression in HEK 293 Cells: An Alternative to E. coli for the Production of Secreted and Intracellular Mammalian Proteins

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Insoluble Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1258))

Abstract

Transient transfection of human embryonic kidney cells (HEK 293) enables the rapid and affordable lab-scale production of recombinant proteins. In this chapter protocols for the expression and purification of both secreted and intracellular proteins using transient expression in HEK 293 cells are described.

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Acknowledgment

The OPPF-UK is funded by the Medical Research Council, UK (grant MR/K018779/1). P.J.W., L.F., and J.W.R.S. are funded by the Wellcome Trust (grants WT085408 and WT100237). J.M.G.C. is funded by a University of Portsmouth IBBS studentship, and S.E.K. by the University of Portsmouth Research Development Fund. M.P., A.U., and S.B. are funded by the BBSRC, UK (grant BB/J008176/1).

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Authors and Affiliations

  1. OPPF-UK, Research Complex at Harwell, R92 Rutherford Appleton Laboratories, Harwell Oxford, Didcot, OX11 0FA, UK

    Joanne E. Nettleship, Nahid Rahman-Huq, Yamini Reddivari & Raymond J. Owens

  2. Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford, OX3 7BN, UK

    Joanne E. Nettleship, Nahid Rahman-Huq, Yamini Reddivari & Raymond J. Owens

  3. Department of Biochemistry, University of Leicester, Henry Wellcome Building, Lancaster Road, Leicester, LE1 9HN, UK

    Peter J. Watson, Louise Fairall & John W. R. Schwabe

  4. Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK

    Mareike G. Posner, Abhishek Upadhyay & Stefan Bagby

  5. Institute of Biomedical and Biomolecular Science, University of Portsmouth, Portsmouth, PO1 2DY, UK

    Jonathan M. G. Chamberlain & Simon E. Kolstoe

Authors
  1. Joanne E. Nettleship
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  2. Peter J. Watson
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  3. Nahid Rahman-Huq
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  4. Louise Fairall
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  5. Mareike G. Posner
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  6. Abhishek Upadhyay
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  7. Yamini Reddivari
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  8. Jonathan M. G. Chamberlain
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  9. Simon E. Kolstoe
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  10. Stefan Bagby
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  11. John W. R. Schwabe
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  12. Raymond J. Owens
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Corresponding author

Correspondence to Joanne E. Nettleship .

Editor information

Editors and Affiliations

  1. Departament de Genètica i de Microbiologia, CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN) and Universitat Autònoma de Barcelona, Bellaterra (Barcelona), Spain

    Elena García-Fruitós

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Nettleship, J.E. et al. (2015). Transient Expression in HEK 293 Cells: An Alternative to E. coli for the Production of Secreted and Intracellular Mammalian Proteins. In: García-Fruitós, E. (eds) Insoluble Proteins. Methods in Molecular Biology, vol 1258. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2205-5_11

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  • DOI: https://doi.org/10.1007/978-1-4939-2205-5_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2204-8

  • Online ISBN: 978-1-4939-2205-5

  • eBook Packages: Springer Protocols

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