title: Expression of human interleukin-2 in recombinant baby hamster kidney, Ltk-, and Chinese hamster ovary cells. Structure of O-linked carbohydrate chains and their location within the polypeptide.
abstract: 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.
authors: Department of Cell Biology and Genetics, Gesellschaft für Biotechnologische Forschung mbH, Braunschweig, Federal Republic of Germany.
authors: H S
authors: HS
authors: Conradt
authors: M
authors: M
authors: Nimtz
authors: K E
authors: KE
authors: Dittmar
authors: W
authors: W
authors: Lindenmaier
authors: J
authors: J
authors: Hoppe
authors: H
authors: H
authors: Hauser
mesh_terms: D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis*; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis / Q000235:genetics*; D010957:Plasmids; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics*; D014162:Transfection
chemical_list: D002241:Carbohydrates; D007376:Interleukin-2; D011994:Recombinant Proteins