Abstract
The production of therapeutic recombinant glycoproteins deals with three main issues: cost, production capacities, and glycosylation. Nowadays, such proteins are expressed in various complex expression systems (CHO, bacteria, etc.); the processes related to those production hosts are time consuming and expensive, or the question of posttranslational modifications (as glycosylation) control is still unresolved. There is a need to find an alternative approach, while maintaining high quality level: the new system must be able to add complex N-glycan structures to proteins of interest. Developed in several strains of Saccharomyces cerevisiae, GlycodExpress™ is an innovative technology that allows production of therapeutic recombinant glycoproteins with humanized and homogeneous N-glycan moieties. We show how to delete mannosyltransferases involved in host N-glycosylation to obtain more than 90% of homogeneity in glycan structures. The methodology developed to select the optimal fusion between a heterologous glycosyl-enzyme and a localization sequences is also presented. Finally, the screening of the best producing strain is illustrated.
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Acknowledgments
C.A., C.B., and C.J. want to thank the entire Glycode team for its high technical and scientific assistance. Special thanks to Ludivine Perrocheau and Jean-Noël Chaize, respectively, head of Biochemistry and Downstream Process Development and head of Fermentation Process for the helping discussions and the critical reading of this manuscript.
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Arico, C., Bonnet, C., Javaud, C. (2013). N-Glycosylation Humanization for Production of Therapeutic Recombinant Glycoproteins in Saccharomyces cerevisiae . In: Beck, A. (eds) Glycosylation Engineering of Biopharmaceuticals. Methods in Molecular Biology, vol 988. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-327-5_4
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DOI: https://doi.org/10.1007/978-1-62703-327-5_4
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