title: Engineering a human-like glycosylation to produce therapeutic glycoproteins based on 6-linked sialylation in CHO cells.
abstract: When recombinant glycoproteins for therapeutic use are to be produced on an industrial scale, there is a crucial need for technologies that can engineer fast-growing stable cells secreting the protein drug at a high rate and with a defined and safe glycosylation profile. Current cell lines approved for drug production are essentially from rodent origin. Their glycosylation machinery often adds undesired carbohydrate determinants which may alter protein folding, induce immunogenicity, and reduce circulatory life span of the drug. Notably, sialic acid as N-acetylneuraminic acid is not efficiently added in most mammalian cells and the 6-linkage is missing in rodent cells. Engineering cells with the various enzymatic activities required for sialic acid transfer has not yet succeeded in providing a human-like pattern of glycoforms to protein drugs. To date, there is a need for engineering animal cells and get highly sialylated products that resemble as closely as possible to human proteins. We have designed ST6Gal minigenes to optimize the ST6GalI sialyltransferase activity and used them to engineer ST6(+)CHO cells. When stably transfected in cells expressing a protein of interest or not, these constructs have proven to equip cell clones with efficient transfer activity of 6-linked sialic acid. In this chapter, we describe a methodology for generating healthy stable adherent clones with hypersialylation activity and high secretion rate.
authors.affiliation: Siamed'Xpress, University Aix-Marseille, Marseille, France.
authors.forename: Nassimal
authors.initials: N
authors.lastname: El Maï
authors.forename: Sandrine
authors.initials: S
authors.lastname: Donadio-Andréi
authors.forename: Chloé
authors.initials: C
authors.lastname: Iss
authors.forename: Valérie
authors.initials: V
authors.lastname: Calabro
authors.forename: Catherine
authors.initials: C
authors.lastname: Ronin
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D011499:Protein Processing, Post-Translational; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D006023:Glycoproteins; D012799:Sialyltransferases