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Enhancing the secretion of recombinant proteins by engineering N-glycosylation sites

✍ Scribed by Yan Liu; Anton Nguyen; Robert L. Wolfert; Shaoqiu Zhuo

Publisher
American Institute of Chemical Engineers
Year
2009
Tongue
English
Weight
294 KB
Volume
25
Category
Article
ISSN
8756-7938

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✦ Synopsis

Abstract

N__‐glycosylation is important for the folding and quality control of membrane and secretory proteins. We used mutagenesis to introduce N‐glycosylation sequons in recombinant proteins to improve their secretion in HEK293 cells. Seven recombinant proteins, with or without endogenous N‐glycosylation sequons, were tested by this method. Our results indicate that N‐glycosylation sequons located at the N‐__ or C‐terminal are glycosylated at high rates and thus the N__‐__ and C‐terminal may be convenient sites for effectively attaching oligosaccharide chains. More importantly, introduction of oligosaccharide chains at such positions has been found to improve the secretion levels for the majority of the recombinant proteins in our studies, regardless of endogenous N__‐__glycosylation, presumably by improving their folding in the endoplasmic reticulum. Β© 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009

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