Engineering Chinese hamster ovary cells to maximize sialic acid content of recombinant glycoproteins
| Autor: | Lynne A. Krummen, David H. Peers, K. Carroll, S. Mehta, Damon I. Papac, Vanessa Chisholm, Nishit B. Modi, Martin Gawlitzek, S. Weikert, S. Tom, Steven M. Chamow, Phillip W. Berman, D. Cowfer, J. Briggs, J. Lofgren, S. Eppler |
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| Rok vydání: | 1999 |
| Předmět: |
Glycosylation
Biomedical Engineering Hamster Bioengineering CHO Cells Applied Microbiology and Biotechnology chemistry.chemical_compound Cricetinae Glycosyltransferase Animals Glycoproteins chemistry.chemical_classification biology Chinese hamster ovary cell Oligosaccharide Galactosyltransferases Molecular biology N-Acetylneuraminic Acid Recombinant Proteins Sialyltransferases Sialic acid carbohydrates (lipids) chemistry Biochemistry Cell culture biology.protein Molecular Medicine Rabbits Genetic Engineering Glycoprotein Biotechnology |
| Zdroj: | Nature Biotechnology. 17:1116-1121 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/15104 |
| Popis: | We have engineered two Chinese hamster ovary cell lines secreting different recombinant glycoproteins to express high levels of human beta1,4-galactosyltransferase (GT, E.C. 2.4.1.38) and/or alpha2, 3-sialyltransferase (ST, E.C. 2.4.99.6). N-linked oligosaccharide structures synthesized by cells overexpressing the glycosyltransferases showed greater homogeneity compared with control cell lines. When GT was overexpressed, oligosaccharides terminating with GlcNAc were significantly reduced compared with controls, whereas overexpression of ST resulted in sialylation of >/=90% of available branches. As expected, GT overexpression resulted in reduction of oligosaccharides terminating with GlcNAc, whereas overexpression of ST resulted in sialylation of >/=90% of available branches. The more highly sialylated glycoproteins had a significantly longer mean residence time in a rabbit model of pharmacokinetics. These experiments demonstrate the feasibility of genetically engineering cell lines to produce therapeutics with desired glycosylation patterns. |
| Databáze: | OpenAIRE |
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