Document Detail


Enhancing glycoprotein sialylation by targeted gene silencing in mammalian cells.
MedLine Citation:
PMID:  20014139     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recombinant glycoproteins produced by mammalian cells represent an important category of therapeutic pharmaceuticals used in human health care. Of the numerous sugars moieties found in glycoproteins, the terminal sialic acid is considered particularly important. Sialic acid has been found to influence the solubility, thermal stability, resistance to protease attack, antigenicity, and specific activity of various glycoproteins. In mammalian cells, it is often desirable to maximize the final sialic acid content of a glycoprotein to ensure its quality and consistency as an effective pharmaceutical. In this study, CHO cells overexpressing recombinant human interferon gamma (hIFNgamma) were treated using short interfering RNA (siRNA) and short-hairpin RNA (shRNA) to reduce expression of two newly identified sialidase genes, Neu1 and Neu3. By knocking down expression of Neu3 we achieved a 98% reduction in sialidase function in CHO cells. The recombinant hIFNgamma was examined for sialic acid content that was found to be increased 33% and 26% respectively with samples from cell stationary phase and death phase as compared to control. Here, we demonstrate an effective targeted gene silencing strategy to enhance protein sialylation using RNA interference (RNAi) technology.
Authors:
Min Zhang; Kerry Koskie; James S Ross; Kevin J Kayser; Matthew V Caple
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  105     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-03-02     Completed Date:  2010-05-25     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1094-105     Citation Subset:  IM    
Copyright Information:
(c) 2009 Wiley Periodicals, Inc.
Affiliation:
Cell Sciences & Development, SAFC Biosciences (a member of Sigma-Aldrich Group), 2909 Laclede Avenue, Saint Louis, MO 63103, USA. zhang_min@lilly.com
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
CHO Cells
COS Cells
Cercopithecus aethiops
Cloning, Molecular
Cricetinae
Cricetulus
Gene Knockdown Techniques
Gene Silencing*
Glycoproteins / biosynthesis*,  metabolism
Humans
Interferon-gamma, Recombinant / biosynthesis,  chemistry,  genetics
Molecular Sequence Data
N-Acetylneuraminic Acid / metabolism*
Neuraminidase / biosynthesis,  genetics*
RNA, Small Interfering / genetics*
Sequence Alignment
Chemical
Reg. No./Substance:
0/Glycoproteins; 0/Interferon-gamma, Recombinant; 0/RNA, Small Interfering; 131-48-6/N-Acetylneuraminic Acid; EC 3.2.1.18/Neuraminidase

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


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