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 modi fications (as glycosylation) control is still unresolved. There is a need tofind 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 TM 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 sequence is also presented.finally, the screening of the best producing strain is illustrated. © Springer Science+Business Media New York 2013.
CITATION STYLE
Arico, C., Bonnet, C., & Javaud, C. (2013). N-glycosylation humanization for production of therapeutic recombinant glycoproteins in saccharomyces cerevisiae. Methods in Molecular Biology, 988, 45–57. https://doi.org/10.1007/978-1-62703-327-5_4
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