Synthesis of Bisected Glycoforms of Recombinant IFN-β by Overexpression of β-1,4-N-Acetylglucosaminyltransferase III in Chinese Hamster Ovary Cells

Genetic engineering of oligosaccharide biosynthesis pathways in mammalian cells makes possible generation of new recombinant glycoproteins of potential importance in the biopharmaceutical industry. Most prior investigations of glycosylation engineering of secreted heterologous glycoproteins involve...

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Bibliographic Details
Published in:Biotechnology progress 1998, Vol.14 (2), p.189-192
Main Authors: Sburlati, Adriana R., Umaña, Pablo, Prati, Elisabetta G. P., Bailey, James E.
Format: Article
Language:English
Subjects:
Animal cell culture
Animals
Biological and medical sciences
Biosynthesis
Biotechnology
CHO Cells
Cricetinae
Culture Media
Enzymes
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Glycoproteins - biosynthesis
Immunoblotting
Interferon-beta - biosynthesis
Methods. Procedures. Technologies
Modification of gene expression level
N-Acetylglucosaminyltransferases - biosynthesis
Oligomers
Polysaccharides
Protein Engineering
Recombinant Proteins - biosynthesis
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Summary:Genetic engineering of oligosaccharide biosynthesis pathways in mammalian cells makes possible generation of new recombinant glycoproteins of potential importance in the biopharmaceutical industry. Most prior investigations of glycosylation engineering of secreted heterologous glycoproteins involve terminal steps of oligosaccharide biosynthesis. In particular, increasing the frequency of bisected structures within the glycoform distribution has not before been considered. A Chinese hamster ovary (CHO) cell line capable of producing bisected oligosaccharides on glycoproteins was created by overexpression of a recombinant N‐acetylglucosaminyltransferase III (GnT‐III) . Interferon β (IFN‐β) was chosen as a model and potential therapeutic secreted heterologous protein to demonstrate the effect of recombinant GnT‐III‐expression on product glycosylation. IFN‐β with bisected oligosaccharides was produced by the GnT‐III‐engineered CHO cells but not by the unmodified parental cell line.
ISSN:8756-7938
1520-6033
DOI:10.1021/bp970118s