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Recombinant Protein Expression in Mammalian Cells pp 179–187Cite as

High Throughput Transfection of HEK293 Cells for Transient Protein Production

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1850))

Abstract

Transient transfection of mammalian cells is used in the biotechnology industry to quickly supply recombinant protein for research and large molecule drug development. Here, we describe a method for high throughput transient transfection of Human Embryonic Kidney 293 (HEK293) cells in 30 mL tubespins using polyethylenimine (PEI) as a transfection reagent. An automated liquid handler can be used to perform pipetting steps for transfecting batches of 96 tubespins, and septa in the tubespin caps allow for rapid processing without decapping. The addition of valproic acid (VPA) to transfection cultures enhances recombinant protein production. The thawing and passaging operations for HEK293 cultures to source the transient transfections are also described.

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Author information

Authors and Affiliations

  1. Department of Early Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA

    Tia A. Arena & Athena W. Wong

  2. Department of Late Stage Cell Culture, Genentech Inc., South San Francisco, CA, USA

    Peter D. Harms

Authors
  1. Tia A. Arena
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  2. Peter D. Harms
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  3. Athena W. Wong
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Corresponding author

Correspondence to Athena W. Wong .

Editor information

Editors and Affiliations

  1. Protein Production and Structure Core Facility (PPSCF), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland

    David L. Hacker

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Arena, T.A., Harms, P.D., Wong, A.W. (2018). High Throughput Transfection of HEK293 Cells for Transient Protein Production. In: Hacker, D. (eds) Recombinant Protein Expression in Mammalian Cells. Methods in Molecular Biology, vol 1850. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8730-6_12

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  • DOI: https://doi.org/10.1007/978-1-4939-8730-6_12

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8729-0

  • Online ISBN: 978-1-4939-8730-6

  • eBook Packages: Springer Protocols

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