DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kwak, Chan-Yeong | ko |
dc.contributor.author | Park, Seung-Yeol | ko |
dc.contributor.author | Lee, Chung Geun | ko |
dc.contributor.author | Okino, Nozomu | ko |
dc.contributor.author | Ito, Makoto | ko |
dc.contributor.author | Kim, Jung Hoe | ko |
dc.date.accessioned | 2017-11-08T05:05:24Z | - |
dc.date.available | 2017-11-08T05:05:24Z | - |
dc.date.created | 2017-10-30 | - |
dc.date.created | 2017-10-30 | - |
dc.date.issued | 2017-10 | - |
dc.identifier.citation | SCIENTIFIC REPORTS, v.7 | - |
dc.identifier.issn | 2045-2322 | - |
dc.identifier.uri | http://hdl.handle.net/10203/226836 | - |
dc.description.abstract | Sialylation regulates the in vivo half-life of recombinant therapeutic glycoproteins, affecting their therapeutic efficacy. Levels of the precursor molecule cytidine monophospho-N-acetylneuraminic acid (CMP-Neu5Ac) are considered a limiting factor in the sialylation of glycoproteins. Here, we show that by reducing the amount of intracellular CMP-Neu5Ac consumed for glycosphingolipid (GSL) biosynthesis, we can increase the sialylation of recombinant human erythropoietin (rhEPO) produced in CHO cells. Initially, we found that treating CHO cells with a potent inhibitor of GSL biosynthesis increases the sialylation of the rhEPO they produce. Then, we established a stable CHO cell line that produces rhEPO in the context of repression of the key GSL biosynthetic enzyme UDP-glucose ceramide glucosyltransferase (UGCG). These UGCG-depleted cells show reduced levels of gangliosides and significantly elevated levels of rhEPO sialylation. Upon further analysis of the resulting N-glycosylation pattern, we discovered that the enhanced rhEPO sialylation could be attributed to a decrease in neutral and mono-sialylated N-glycans and an increase in di-sialylated N-glycans. Our results suggest that the therapeutic efficacy of rhEPO produced in CHO cells can be improved by shunting intracellular CMP-Neu5Ac away from GSL biosynthesis and toward glycoprotein sialylation. | - |
dc.language | English | - |
dc.publisher | NATURE PUBLISHING GROUP | - |
dc.subject | HAMSTER OVARY CELLS | - |
dc.subject | GROWTH-FACTOR RECEPTOR | - |
dc.subject | SIALIC-ACID CONTENT | - |
dc.subject | GLUCOSYLCERAMIDE SYNTHASE | - |
dc.subject | INTERFERON-GAMMA | - |
dc.subject | INSECT CELLS | - |
dc.subject | GLYCOSYLATION | - |
dc.subject | ERYTHROPOIETIN | - |
dc.subject | PROTEIN | - |
dc.subject | EXPRESSION | - |
dc.title | Enhancing the sialylation of recombinant EPO produced in CHO cells via the inhibition of glycosphingolipid biosynthesis | - |
dc.type | Article | - |
dc.identifier.wosid | 000412950600050 | - |
dc.identifier.scopusid | 2-s2.0-85031300343 | - |
dc.type.rims | ART | - |
dc.citation.volume | 7 | - |
dc.citation.publicationname | SCIENTIFIC REPORTS | - |
dc.identifier.doi | 10.1038/s41598-017-13609-4 | - |
dc.contributor.localauthor | Kim, Jung Hoe | - |
dc.contributor.nonIdAuthor | Park, Seung-Yeol | - |
dc.contributor.nonIdAuthor | Okino, Nozomu | - |
dc.contributor.nonIdAuthor | Ito, Makoto | - |
dc.description.isOpenAccess | Y | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | GROWTH-FACTOR RECEPTOR | - |
dc.subject.keywordPlus | SIALIC-ACID CONTENT | - |
dc.subject.keywordPlus | GLUCOSYLCERAMIDE SYNTHASE | - |
dc.subject.keywordPlus | INTERFERON-GAMMA | - |
dc.subject.keywordPlus | INSECT CELLS | - |
dc.subject.keywordPlus | GLYCOSYLATION | - |
dc.subject.keywordPlus | ERYTHROPOIETIN | - |
dc.subject.keywordPlus | PROTEIN | - |
dc.subject.keywordPlus | EXPRESSION | - |
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