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Rapid Antibody Glycoengineering in CHO Cells Via RNA Interference and CGE-LIF N-Glycomics

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Glycosylation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2370))

Abstract

The impact of the glycan distribution on the in vivo function and half-life of monoclonal antibodies has long motivated the genetic engineering of producer cells to achieve structures that enhance efficacy, safety and stability. To facilitate glycoengineering of IgG-producing Chinese hamster ovary cells, we present a rapid protocol that involves the use of RNA interference for the knockdown of genes of interest coupled with capillary gel electrophoresis and laser-induced fluorescence detection (CGE-LIF) for fast, high-throughput glycan analysis. We apply this methodology to the Fut8 gene, responsible for the addition of core fucose, which is a typical target for increasing antibody-dependent cellular cytotoxicity.

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Acknowledgments

PK thanks the Department of Chemical Engineering, Imperial College London, for his scholarship. RD thanks the U.K. Biotechnology and Biological Sciences Research Council for his studentship. MM is funded by the U.K. Biotechnology and Biological Sciences Research Council (Grant reference: BB/S006206/1). IAG thanks the Irish Research Council (Scholarship No. GOIPG/2017/1049) and CONACyT (Scholarship No. 438330).

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Authors and Affiliations

  1. Department of Chemical Engineering, Imperial College London, London, UK

    Pavlos Kotidis, Masue Marbiah, Roberto Donini, Karen M. Polizzi & Cleo Kontoravdi

  2. Imperial College Centre for Synthetic Biology, Imperial College London, London, UK

    Masue Marbiah & Karen M. Polizzi

  3. Department of Life Sciences, Imperial College London, London, UK

    Roberto Donini & Stuart M. Haslam

  4. School of Chemical & Bioprocess Engineering, University College Dublin, Dublin, Ireland

    Itzcóatl A. Gómez & Ioscani Jimenez del Val

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  1. Pavlos Kotidis
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  2. Masue Marbiah
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  4. Itzcóatl A. Gómez
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  8. Cleo Kontoravdi
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Correspondence to Cleo Kontoravdi .

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  1. School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland

    Gavin P. Davey

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Kotidis, P. et al. (2022). Rapid Antibody Glycoengineering in CHO Cells Via RNA Interference and CGE-LIF N-Glycomics. In: Davey, G.P. (eds) Glycosylation. Methods in Molecular Biology, vol 2370. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1685-7_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1685-7_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1684-0

  • Online ISBN: 978-1-0716-1685-7

  • eBook Packages: Springer Protocols

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