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Generation of Recombinant N-Linked Glycoproteins in E. coli

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Heterologous Gene Expression in E.coli

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1586))

Abstract

The production of N-linked recombinant glycoproteins is possible in a variety of biotechnology host cells, and more recently in the bacterial workhorse, Escherichia coli. This methods chapter will outline the components and procedures needed to produce N-linked glycoproteins in E. coli, utilizing Campylobacter jejuni glycosylation machinery, although other related genes can be used with minimal tweaks to this methodology. To ensure a successful outcome, various methods will be highlighted that can confirm glycoprotein production to a high degree of confidence, including the gold standard of mass spectrometry analysis.

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Acknowledgments

The authors acknowledge and are deeply thankful for funding from the UK’s Biotechnology and Biological Sciences Research Council (BBSRC) through the Bioprocess Research Industry Club (BRIC) programme (BB/K011200/1 and BB/F004842/1).

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, ChELSI Institute, University of Sheffield, Mappin Street, Sheffield, S1 3JD, UK

    Benjamin Strutton, Stephen R. P. Jaffé & Jagroop Pandhal

  2. Faculty of Science, Agriculture and Engineering, Newcastle University, Newcastle Upon Tyne, UK

    Phillip C. Wright

Authors
  1. Benjamin Strutton
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  2. Stephen R. P. Jaffé
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  3. Jagroop Pandhal
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  4. Phillip C. Wright
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Corresponding author

Correspondence to Jagroop Pandhal .

Editor information

Editors and Affiliations

  1. SGC, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom

    Nicola A. Burgess-Brown

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Strutton, B., Jaffé, S.R.P., Pandhal, J., Wright, P.C. (2017). Generation of Recombinant N-Linked Glycoproteins in E. coli . In: Burgess-Brown, N. (eds) Heterologous Gene Expression in E.coli. Methods in Molecular Biology, vol 1586. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6887-9_15

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  • DOI: https://doi.org/10.1007/978-1-4939-6887-9_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6885-5

  • Online ISBN: 978-1-4939-6887-9

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