Title : Expression of human
interleukin-2 in recombinant baby hamster kidney,
Ltk-, and Chinese hamster ovary cells
Abstract :
- Structure of O-linked carbohydrate chains and their location within the polypeptide
- The similarity or identity of O-glycosylation in glycoproteins from natural sources or produced in heterologous cell lines, a central problem for the development of many biotechnologically relevant production processes , was examined using interleukin-2 ( IL-2 ) as a model
- Human interleukin-2 was constitutively expressed in several mammalian cell lines in high amounts
- The recombinant proteins were purified to homogeneity and their carbohydrate structures were analyzed
- Only the NeuAc alpha 2-3Gal beta 1-3[NeuAc alpha 2-6]GalNAc oligosaccharide structure or the NeuAc alpha 2-3Gal beta 1-3GalNAc were found in all IL-2 preparations secreted from recombinant Ltk-, Chinese hamster ovary, and baby hamster kidney cell lines
- The O-linked chains were exclusively linked to Thr in position 3 of the polypeptide chain which is the carbohydrate attachment site in natural human IL-2
- The proportions of O-glycosylated versus nonglycosylated forms of the protein secreted by each recombinant cell line were independent of productivity or of cell culture conditions
- Our results show that O-glycosylated human IL-2 can be produced by applying recombinant DNA technology in heterologous cell lines with the same type of post-translational modification that is observed for the protein secreted from natural T lymphocytes