Engineered CHO cells for production of diverse, homogeneous glycoproteins (original) (raw)

Nature Biotechnology volume 33, pages 842–844 (2015)Cite this article

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Abstract

Production of glycoprotein therapeutics in Chinese hamster ovary (CHO) cells is limited by the cells' generic capacity for _N-_glycosylation, and production of glycoproteins with desirable homogeneous glycoforms remains a challenge. We conducted a comprehensive knockout screen of glycosyltransferase genes controlling _N-_glycosylation in CHO cells and constructed a design matrix that facilitates the generation of desired glycosylation, such as human-like α2,6-linked sialic acid capping. This engineering approach will aid the production of glycoproteins with improved properties and therapeutic potential.

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Acknowledgements

We would like to express our sincere gratitude to the SAFC Sigma team including K. Kayser and N. Sealover for their help with ZFN targeting constructs. We are also grateful to B. Palsson for help throughout this work and critical comments on the manuscript. We are grateful to M. Uhlen, L.E. Pedersen and B. Voldborg at the Novo Nordisk Foundation Center for Biosustainability, Danish Technical University, for RNA-seq analysis. This work was supported by the Novo Nordisk Foundation, Kirsten og Freddy Johansen Fonden, The Mizutani Foundation, A.P. Møller og Hustru Chastine Mc-Kinney Møllers Fond til Almene Formaal, Copenhagen University Excellence Programme for Interdisciplinary Research (CDO2016), the Danish National Research Foundation (DNRF107) and The Danish Councils for Strategic and Independent Research. All reagents and cell lines used in the study are available upon request for research purposes under a material transfer agreement.

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Authors and Affiliations

  1. Center for Glycomics, Cellular and Molecular Medicine and Odontology, Faculty of Health Sciences, University of Copenhagen, Copenhagen N, Denmark
    Zhang Yang, Shengjun Wang, Adnan Halim, Morten Alder Schulz, Shamim H Rahman, Malene B Vester-Christensen, Claus Kristensen, Sergey Y Vakhrushev, Eric Paul Bennett, Hans H Wandall & Henrik Clausen
  2. The Novo Nordisk Foundation Center for Biosustainability, The Danish Technical University, Denmark
    Zhang Yang, Shengjun Wang, Adnan Halim & Henrik Clausen
  3. Biotech Research and Innovation Centre, University of Copenhagen, Copenhagen, Denmark
    Morten Frodin
  4. Novo Nordisk A/S, Måløv, Denmark
    Shamim H Rahman, Malene B Vester-Christensen & Carsten Behrens

Authors

  1. Zhang Yang
  2. Shengjun Wang
  3. Adnan Halim
  4. Morten Alder Schulz
  5. Morten Frodin
  6. Shamim H Rahman
  7. Malene B Vester-Christensen
  8. Carsten Behrens
  9. Claus Kristensen
  10. Sergey Y Vakhrushev
  11. Eric Paul Bennett
  12. Hans H Wandall
  13. Henrik Clausen

Contributions

Z.Y. designed, planned and ran most of the ZFN targeting experiments and co-wrote the manuscript. S.W. performed purification of EPO and IgG. A.H. performed the glycoprofiling. M.A.S. contributed to expression of EPO and IgG. M.F. contributed to ZFN design. S.H.R., C.B. and M.B.V.-C. contributed to the design of experiments for homogeneous glycoPEGylation of monoantennary _N-_glycans. S.Y.V., E.P.B., C.K. and H.H.W. contributed to the design of experiments. H.C. designed, planned and co-wrote the manuscript.

Corresponding authors

Correspondence toZhang Yang or Henrik Clausen.

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Competing interests

A patent application has been filed on which some of the authors are listed as inventors.

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Yang, Z., Wang, S., Halim, A. et al. Engineered CHO cells for production of diverse, homogeneous glycoproteins.Nat Biotechnol 33, 842–844 (2015). https://doi.org/10.1038/nbt.3280

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