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High-Throughput Insect Cell Protein Expression Applications

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High Throughput Protein Expression and Purification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 498))

Summary

The Baculovirus Expression Vector System (BEVS) is one of the most efficient systems for production of recombinant proteins and consequently its application is wide-spread in industry as well as in academia. Since the early 1970s, when the first stable insect cell lines were established and the infectivity of bacu-lovirus in an in vitro culture system was demonstrated (1, 2), virtually thousands of reports have been published on the successful expression of proteins using this system as well as on method improvement. However, despite its popularity the system is labor intensive and time consuming. Moreover, adaptation of the system to multi-parallel (high-throughput) expression is much more difficult to achieve than with E. coli due to its far more complex nature. However, recent years have seen the development of strategies that have greatly enhanced the stream-lining and speed of baculovirus protein expression for increased throughput via use of automation and miniaturization. This chapter therefore tries to collate these developments in a series of protocols (which are modifications to standard procedure plus several new approaches) that will allow the user to expedite the speed and throughput of baculovirus-mediated protein expression and facilitate true multi-parallel, high-throughput protein expression profiling in insect cells. In addition we also provide a series of optimized protocols for small and large-scale transient insect cell expression that allow for both the rapid analysis of multiple constructs and the concomitant scale-up of those selected for on-going analysis. Since this approach is independent of viral propagation, the timelines for this approach are markedly shorter and offer a significant advantage over standard bacu-lovirus expression approach strategies in the context of HT applications.

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Acknowledgments

The authors wish to thank Brendan Kerins and Jean-Marc Schlaeppi for technical support and James Groarke for useful discussions and comments during the preparation of this manuscript. This manuscript is dedicated to the memory of Robert Cheung

Author information

Authors and Affiliations

  1. Biologics Center, Novartis Institutes for Biomedical Research, Basel, Switzerland

    Mirjam Buchs, Yann Pouliquen, Sabine Geisse & Marion Mahnke

  2. Protein Structure Unit, Centre for Proteomic Chemistry, Novartis Institutes for Biomedical Research, Cambridge, MA, USA

    Ernie Kim & Ian Hunt

Authors
  1. Mirjam Buchs
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  2. Ernie Kim
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  3. Yann Pouliquen
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  4. Michael Sachs
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  5. Sabine Geisse
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  6. Marion Mahnke
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  7. Ian Hunt
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Editor information

Editors and Affiliations

  1. Glaxo Smith Kline, Walnut Creek, CA, USA

    Sharon A. Doyle

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Buchs, M. et al. (2009). High-Throughput Insect Cell Protein Expression Applications. In: Doyle, S.A. (eds) High Throughput Protein Expression and Purification. Methods in Molecular Biology, vol 498. Humana Press. https://doi.org/10.1007/978-1-59745-196-3_14

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  • DOI: https://doi.org/10.1007/978-1-59745-196-3_14

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-879-9

  • Online ISBN: 978-1-59745-196-3

  • eBook Packages: Springer Protocols

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