title: N-glycosylation humanization for production of therapeutic recombinant glycoproteins in Saccharomyces cerevisiae.
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.
authors.affiliation: Glycode SAS, Uzerche, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Arico
authors.forename: Christine
authors.initials: C
authors.lastname: Bonnet
authors.forename: Christophe
authors.initials: C
authors.lastname: Javaud
mesh_terms: D002240:Carbohydrate Sequence; D017353:Gene Deletion; D006023:Glycoproteins / Q000096:biosynthesis*; D006031:Glycosylation; D006801:Humans; D008364:Mannosyltransferases / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D006023:Glycoproteins; D008562:Membrane Glycoproteins; C075636:OCH1 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C086604:MNN1 protein, S cerevisiae; D008364:Mannosyltransferases