| Metabolic control of recombinant protein N-glycan processing in NS0 and CHO cells. | |
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MedLine Citation:
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PMID: 11257601 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Chinese hamster ovary and murine myeloma NS0 cells are currently favored host cell types for the production of therapeutic recombinant proteins. In this study, we compared N-glycan processing in GS-NS0 and GS-CHO cells producing the same model recombinant glycoprotein, tissue inhibitor of metalloproteinases 1. By manipulation of intracellular nucleotide-sugar content, we examined the feasibility of implementing metabolic control strategies aimed at reducing the occurrence of murine-specific glycan motifs on NS0-derived recombinant proteins, such as Galalpha1,3Galbeta1,4GlcNAc. Although both CHO and NS0-derived oligosaccharides were predominantly of the standard complex type with variable sialylation, 30% of N-glycan antennae associated with NS0-derived TIMP-1 terminated in alpha1,3-linked galactose residues. Furthermore, NS0 cells conferred a greater proportion of terminal N-glycolylneuraminic (sialic) acid residues as compared with the N-acetylneuraminic acid variant. Inclusion of the nucleotide-sugar precursors, glucosamine (10 mM, plus 2 mM uridine) and N-acetylmannosamine (20 mM), in culture media were shown to significantly increase the intracellular pools of UDP-N-acetylhexosamine and CMP-sialic acid, respectively, in both NS0 and CHO cells. The elevated UDP-N-acetylhexosamine content induced by the glucosamine/uridine treatment was associated with an increase in the antennarity of N-glycans associated with TIMP-1 produced in CHO cells but not N-glycans associated with TIMP-1 from NS0 cells. In addition, elevated UDP-N-acetylhexosamine content was associated with a slight decrease in sialylation in both cell lines. The elevated CMP-sialic acid content induced by N-acetylmannosamine had no effect on the overall level of sialylation of TIMP-1 produced by both CHO and NS0 cells, although the ratio of N-glycolylneuraminic acid:N-acetylneuraminic acid associated with NS0-derived TIMP-1 changed from 1:1 to 1:2. These data suggest that manipulation of nucleotide-sugar metabolism can promote changes in N-glycan processing that are either conserved between NS0 and CHO cells or specific to either NS0 cells or CHO cells. |
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Authors:
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K N Baker; M H Rendall; A E Hills; M Hoare; R B Freedman; D C James |
Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Biotechnology and bioengineering Volume: 73 ISSN: 0006-3592 ISO Abbreviation: Biotechnol. Bioeng. Publication Date: 2001 May |
Date Detail:
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Created Date: 2001-03-21 Completed Date: 2001-06-14 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7502021 Medline TA: Biotechnol Bioeng Country: United States |
Other Details:
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Languages: eng Pagination: 188-202 Citation Subset: IM |
Copyright Information:
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Copyright 2001 John Wiley & Sons, Inc. |
Affiliation:
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Department of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Bioreactors CHO Cells Cell Division / drug effects Chromatography, High Pressure Liquid Cricetinae Galactose / metabolism Glucosamine / pharmacology Glycosylation / drug effects Hexosamines / pharmacology Mice N-Acetylneuraminic Acid / metabolism Nucleotides / metabolism Polysaccharides / chemistry, metabolism* Recombinant Proteins / metabolism Sialyltransferases / metabolism Tissue Inhibitor of Metalloproteinase-1 / chemistry, metabolism* Tumor Cells, Cultured |
| Chemical | |
Reg. No./Substance:
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0/Hexosamines; 0/Nucleotides; 0/Polysaccharides; 0/Recombinant Proteins; 0/Tissue Inhibitor of Metalloproteinase-1; 131-48-6/N-Acetylneuraminic Acid; 26566-61-0/Galactose; 3416-24-8/Glucosamine; 4773-29-9/N-acetylmannosamine; EC 2.4.99.-/Sialyltransferases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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