Document Detail


Improved glycosylation of a foreign protein by Tn-5B1-4 cells engineered to express mammalian glycosyltransferases.
MedLine Citation:
PMID:  11400096     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The major advantages of using the baculovirus-insect cell system for recombinant protein production are its ability to produce large amounts of recombinant proteins and its ability to provide eucaryotic modifications, such as glycosylation. However, the glycans linked to recombinant glycoproteins produced by this system typically differ from those found on native mammalian products. This is an important problem because glycans on mammalian glycoproteins can influence their functions in many different ways. The inability of baculovirus-infected insect cells to produce glycans identical to those found on native mammalian glycoproteins is due, in part, to the absence of functional levels of certain glycosyltransferases in insect cells. Thus, the purpose of this study was to engineer these activities into Tn-5B1-4, an established insect cell line that is widely used as a host for baculovirus-mediated protein production. Expression plasmids were constructed in which cDNAs encoding mammalian beta1,4-galactosyltransferase and alpha2,6-sialyltransferase were placed under the transcriptional control of a baculovirus immediate early promoter. These plasmids were then used to isolate two different transgenic Tn-5B1-4 derivatives and the biological and biochemical properties of these cell lines were examined. The results show that both of the engineered insect cell lines have improved glycoprotein-processing capabilities, relative to the parental cell line.
Authors:
K Breitbach; D L Jarvis
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  74     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-06-11     Completed Date:  2001-10-04     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  230-9     Citation Subset:  IM    
Copyright Information:
Copyright 2001 John Wiley & Sons, Inc.
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MeSH Terms
Descriptor/Qualifier:
Animals
Anti-Bacterial Agents / pharmacology
Baculoviridae / genetics
Cells, Cultured
Glycosylation
Glycosyltransferases / genetics*,  metabolism
Mammals / genetics*
Moths / cytology*
N-Acetyllactosamine Synthase / genetics,  metabolism
Neomycin / pharmacology
Plasmids / drug effects
Protein Engineering / methods*
Recombinant Proteins / genetics,  metabolism
Sialyltransferases / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
GM49734/GM/NIGMS NIH HHS; R01 GM049734/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Anti-Bacterial Agents; 0/Recombinant Proteins; 1404-04-2/Neomycin; EC 2.4.-/Glycosyltransferases; EC 2.4.1.90/N-Acetyllactosamine Synthase; EC 2.4.99.-/Sialyltransferases; EC 2.4.99.1/beta-D-galactoside alpha 2-6-sialyltransferase
Comments/Corrections

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