Document Detail


Vaccinia virus-based expression of gp120 and EGFP: survey of mammalian host cell lines.
MedLine Citation:
PMID:  15903257     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Production of recombinant proteins with the vaccinia virus expression system in five mammalian cell lines (HeLa, BS-C-1, Vero, MRC-5, and 293) was investigated for protein yield and proper posttranslational modifications. Regulatory acceptance of the host cell line was taken into consideration, where Vero, MRC-5, and 293 were considered more acceptable to the regulatory authorities. Relevant process knowledge for ease of scale-up with the particular cell type was also considered. Two proteins were expressed, enhanced green fluorescent protein (EGFP) in the cytoplasm and gp120, an HIV envelope coat protein that is secreted into the culture medium. HeLa cells produced the most EGFP at 17.2 microg/well with BS-C-1 and 293 following. BS-C-1 produced the most gp120 at 28.2 microg/mL with 293 and Vero following. Therefore, of the three most appropriate cell lines (Vero, MRC-5, and 293) for production processes, the best results were obtained with 293 cells. Although MRC-5 had a very high productivity on a per cell basis, the low cell density and slow growth rate made the overall production insufficient. Because gp120 contained a significant amount of posttranslational modification, this protein, produced by the different cell lines, was further analyzed by PNGase digestion suggesting N-linked glycosylation modifications in all cell lines tested. On the basis of these results and overall process considerations, 293 cells are recommended for further production process optimization in a serum-free suspension system.
Authors:
Nicole A Bleckwenn; William E Bentley; Joseph Shiloach
Related Documents :
19172057 - In vitro culture of various species of microsporidia causing keratitis: evaluation of t...
17537857 - Enterovirus 70 receptor utilization is controlled by capsid residues that also regulate...
8397027 - Human papillomavirus type 18 dna in so-called hep-2, kb and fl cells--further evidence ...
22447857 - The wnt inhibitor dickkopf 1 and bone morphogenetic protein 4 rescue adipogenesis in hy...
21715087 - Phytochelatin synthase activity as a marker of metal pollution.
19067977 - The timely deposition of callose is essential for cytokinesis in arabidopsis.
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biotechnology progress     Volume:  21     ISSN:  8756-7938     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2005 Jan-Feb
Date Detail:
Created Date:  2005-05-20     Completed Date:  2005-07-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  186-91     Citation Subset:  IM    
Affiliation:
Biotechnology Unit, NIDDK, National Institutes of Health, DHHS, Bldg 14A Rm 173, 9000 Rockville Pike, Bethesda, Maryland 20892, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Cell Culture Techniques / methods
Cell Line
Cells, Cultured
Cercopithecus aethiops
Green Fluorescent Proteins / biosynthesis*,  genetics
HIV Envelope Protein gp120 / analysis,  biosynthesis*,  genetics
Hela Cells
Humans
Sensitivity and Specificity
Time Factors
Vaccinia virus / genetics,  metabolism*
Vero Cells
Chemical
Reg. No./Substance:
0/HIV Envelope Protein gp120; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins

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


Previous Document:  Quorum-sensing-based toolbox for regulatable transgene and siRNA expression in mammalian cells.
Next Document:  Inflammatory response and apoptosis in newborn lungs after meconium aspiration.