Document Detail


Sialylation enhancement of CTLA4-Ig fusion protein in Chinese hamster ovary cells by dexamethasone.
MedLine Citation:
PMID:  20521303     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The importance of glycoprotein sialic acid levels is well known, as increased levels have been shown to increase in vivo serum half-life profiles. Here we demonstrate for the first time that dexamethasone (DEX) was capable of improving the sialylation of a CTLA4-Ig fusion protein produced by Chinese hamster ovary (CHO) cells. DEX was shown to enhance the intracellular addition of sialic acid by sialyltransferases as well as reduce extracellular removal of sialic acid by sialidase cleavage. We illustrated that DEX addition resulted in increased expression of the glycosyltransferases alpha2,3-sialyltransferase (alpha2,3-ST) and beta1,4-galactosyltransferase (beta1,4-GT) in CHO cells. Based upon our previous results showing DEX addition increased culture cell viability, we confirmed here that cultures treated with DEX also resulted in decreased sialidase activity. Addition of the glucocorticoid receptor (GR) antagonist mifepristone (RU-486) was capable of blocking the increase in sialylation by DEX which further supports that DEX affected sialylation as well as provides evidence that the sialylation enhancement effects of DEX on recombinant CHO cells occurred through the GR. Finally, the effects of DEX on increasing sialylation were then confirmed in 5-L controlled bioreactors. Addition of 1 microM DEX to the bioreactors on day 2 resulted in harvests with average increases of 16.2% for total sialic acid content and 15.8% in the protein fraction with N-linked sialylation. DEX was found to be a simple and effective method for increasing sialylation of this CTLA4-Ig fusion protein expressed in CHO cells.
Authors:
Ying Jing; Yueming Qian; Zheng Jian Li
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  107     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 Oct 
Date Detail:
Created Date:  2010-09-02     Completed Date:  2010-12-08     Revised Date:  2013-05-24    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  488-96     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Wiley Periodicals, Inc.
Affiliation:
Biologics Process and Product Development, Technical Operations, Bristol-Myers Squibb Company, East Syracuse, New York 13057, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CHO Cells
Cricetinae
Cricetulus
Dexamethasone / metabolism*
Gene Expression / drug effects
Immunoconjugates / metabolism*
Mifepristone / metabolism
N-Acetylneuraminic Acid / metabolism*
Neuraminidase / metabolism
Receptors, Glucocorticoid / antagonists & inhibitors
Sialyltransferases / metabolism
Chemical
Reg. No./Substance:
0/Immunoconjugates; 0/Receptors, Glucocorticoid; 131-48-6/N-Acetylneuraminic Acid; 50-02-2/Dexamethasone; 7D0YB67S97/abatacept; 84371-65-3/Mifepristone; EC 2.4.99.-/Sialyltransferases; EC 3.2.1.18/Neuraminidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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