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A High-Throughput System for Transient and Stable Protein Production in Mammalian Cells

Part of the Methods in Molecular Biology book series (MIMB,volume 2025)


Recombinant protein expression and purification is an essential component of biomedical research and drug discovery. Advances in automation and laboratory robotics have enabled the development of highly parallel and rapid processes for cell culture and protein expression, purification, and analysis. Human embryonic kidney (HEK) cells and Chinese hamster ovary (CHO) cells have emerged as the standard host cell workhorses for producing recombinant secreted mammalian proteins by using both transient and stable production strategies. In this chapter we describe a fully automated custom platform, Protein Expression and Purification Platform (PEPP), used for transient protein production from HEK cells and stable protein production from CHO cells. Central to PEPP operation is a suite of custom robotic and instrumentation platforms designed and built at GNF, custom cell culture ware, and custom scheduling software referred to as Runtime. The PEPP platform enables cost-effective, facile, consistent production of proteins at quantities and quality useful for early stage drug discovery tasks such as screening, bioassays, protein engineering, and analytics.

Key words

  • Human embryonic kidney
  • HEK
  • Chinese hamster ovary
  • CHO
  • Mammalian
  • Protein
  • Expression
  • Protein purification
  • Transient
  • Stable
  • Cell culture
  • Automation
  • Robotics
  • PEPP

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  • DOI: 10.1007/978-1-4939-9624-7_5
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The authors would like to acknowledge Salvatore Fanale, Analisa Benedetto, Heath Klock, Julie Vance, Melisa Low, Sarah Cox, Daniel McMullan, Mark Knuth, Marc Gustafson, Jim Chang, Marie Smith, Daniel Sipes, and James Mainquist for their contributions to the protocols and technologies described in this chapter.

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Correspondence to Sarah M. Rue .

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Rue, S.M., Anderson, P.W., Gaylord, M.R., Miller, J.J., Glaser, S.M., Lesley, S.A. (2019). A High-Throughput System for Transient and Stable Protein Production in Mammalian Cells. In: Vincentelli, R. (eds) High-Throughput Protein Production and Purification. Methods in Molecular Biology, vol 2025. Humana, New York, NY.

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  • Print ISBN: 978-1-4939-9623-0

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