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Large-Scale Transient Expression of Therapeutic Proteins in Mammalian Cells

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Therapeutic Proteins

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 308))

Abstract

Research of the molecular and cellular biology of mammalian cells would be highly restricted if not for the development of methods to deliver exogenous DNA into cultivated cells. Transient transfection of mammalian cells became a routine research tool following a landmark publication by Graham and Van der Eb, who presented the calcium phosphate method as an assay to test the infectivity of purified viral DNA in 1973 (1). For the first time, this technique allowed the delivery of genes into animal cells without the help of a virus. Novel techniques for nonviral gene transfer have been developed and improved since then. Today, numerous commercial transfection agents for a wide range of cell lines are available (2). This chapter focuses on the two leading methods for large-scale applications of transient gene expression (TGE): calcium phosphate DNA coprecipitates (3,4) and polyethylenimine (PEI)-DNA polyplexes (5,6). Although calcium phosphate DNA coprecipitation is a well-established method that has been modified for high efficiency by many authors (7–12), PEI has only recently been developed as a transfection vehicle (13–15), but it has achieved rapid acceptance for large-scale suspension transfections because of its simplicity in handling and its efficacy of gene transfer to many different cell types.

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Author information

Authors and Affiliations

  1. Novartis Pharma Research CT/BMP, Basel, Switzerland

    Sabine Geisse

  2. Laboratory of Cellular Biotechnology, SV-IGBB-LBTC, EPFL, Lausanne, Switzerland

    Martin Jordan & Florian M. Wurm

Authors
  1. Sabine Geisse
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  2. Martin Jordan
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  3. Florian M. Wurm
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Editor information

Editors and Affiliations

  1. Protein Science Group, Department of Biosciences, University of Kent, Canterbury, Kent, UK

    C. Mark Smales

  2. School of Engineering, University of Queensland, St. Lucia, Queensland, Australia

    David C. James

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© 2005 Humana Press Inc.

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Geisse, S., Jordan, M., Wurm, F.M. (2005). Large-Scale Transient Expression of Therapeutic Proteins in Mammalian Cells. In: Smales, C.M., James, D.C. (eds) Therapeutic Proteins. Methods in Molecular Biology™, vol 308. Humana Press. https://doi.org/10.1385/1-59259-922-2:087

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  • DOI: https://doi.org/10.1385/1-59259-922-2:087

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-390-9

  • Online ISBN: 978-1-59259-922-6

  • eBook Packages: Springer Protocols

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