Document Detail


Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines.
MedLine Citation:
PMID:  1367489     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recombinant Autographa california baculoviruses expressing genes for pseudorabies virus glycoprotein (gp50T), human plasminogen (HPg), and beta-galactosidase (beta-gal) were used to infect 23 cell lines or strains. The objectives were to compare amounts of recombinant proteins expressed in the cell lines, compare yields from clones and parent lines, investigate the effects of long-term culture in serum-free medium on production, and determine if some lines yield gp50T with different glycosylation patterns. For HPg, IZD-MB0503 had the highest yield and four other lines (IPLB-TN-R2, IPLB-SF-1254, IPLB-LdEIta, and CM-1) had levels above that of SF-9 cells. For gp50T, four lines (IPLB-HvT1, IPLB-SF21AE, IPLB-SF21AE-15, and IPLB-SF-1254) had higher amounts than SF-9 cells. Some lines yielded gp50T with molecular mass about 1000 daltons larger than that from SF-9 cells, which suggests increased oligosaccharide processing. Equally high levels of beta-gal were expressed in three lines (SF-9, IZD-MB0503, and BCIRL-PX2-HNV3). The major conclusion is that no single cell line produced highest yields for all three recombinant proteins. Four lines were cultured in serum-free medium for 31-34 passages and then infected with the three recombinant viruses. For most cell line-recombinant combinations, the yields in serum-free medium were equal to or better than those in serum-supplemented medium. Medium composition had a much stronger effect on foreign gene expression than on susceptibility of cells to wild-type virus.
Authors:
W F Hink; D R Thomsen; D J Davidson; A L Meyer; F J Castellino
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Biotechnology progress     Volume:  7     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    1991 Jan-Feb
Date Detail:
Created Date:  1991-05-21     Completed Date:  1991-05-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  9-14     Citation Subset:  B    
Affiliation:
Department of Entomology, Ohio State University, Columbus 43210.
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MeSH Terms
Descriptor/Qualifier:
Animals
Baculoviridae / genetics*
Blood
Cell Line
Clone Cells
Culture Media
Gene Expression*
Genetic Vectors*
Glycosylation
Humans
Insects / metabolism*
Plasminogen / biosynthesis,  genetics
Recombinant Proteins / genetics*
Transfection
Viral Envelope Proteins / biosynthesis,  genetics
beta-Galactosidase / biosynthesis,  genetics
Chemical
Reg. No./Substance:
0/Culture Media; 0/Recombinant Proteins; 0/Viral Envelope Proteins; 0/pseudorabies virus glycoproteins; 9001-91-6/Plasminogen; EC 3.2.1.23/beta-Galactosidase

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


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