| Expression of SEAP (secreted alkaline phosphatase) by baculovirus mediated transduction of HEK 293 cells in a hollow fiber bioreactor system. | |
| | |
MedLine Citation:
|
PMID: 18499293 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
A BacMam baculovirus was designed in our laboratory to express the reporter protein secreted alkaline phosphatase (SEAP) driven by the immediate early promoter of human cytomegalovirus promoter (CMV). In vitro tests have been carried out using this recombinant baculovirus to study the secreted protein in two cell lines and under various culture conditions. The transductions were carried out on two commonly used mammalian cell lines namely the human embryonic kidney (HEK 293A) and Chinese hamster ovary (CHO-K1). Initial studies clearly demonstrated that the transient expression of SEAP was at least 10-fold higher in the HEK 293 cells than the CHO cells under equivalent experimental conditions. Factorial design experiments were done to study the effect of different parameters such as cell density, MOI, and the histone deacetylase inhibitor, trichostatin A concentration. The multiplicity of infection (MOI) and the cell density were found to have the most impact on the process. The enhancer trichostatin A also showed some positive effect. The production of secreted protein in a batch reactor was studied using the Wave disposable bioreactor system. A semi-continuous perfusion process was developed to extend the period of gene expression in mammalian cells using a hollow fiber bioreactor system (HFBR). The growth of cells and viability in both systems was monitored by offline analyses of metabolites. The expression of recombinant protein could be maintained over an extended period of time up to 30 days in the HFBR. |
| | |
Authors:
|
B A Jardin; Y Zhao; M Selvaraj; J Montes; R Tran; S Prakash; C B Elias |
Related Documents
:
|
7244653 - Diameter of the cell-to-cell junctional membrane channels as probed with neutral molecu... 15199233 - High-resolution capillary electrophoresis of nitrite and nitrate in biological samples. 9930073 - The limited difference between keratin patterns of squamous cell carcinomas and adenoca... |
Publication Detail:
|
Type: Journal Article Date: 2008-05-21 |
Journal Detail:
|
Title: Journal of biotechnology Volume: 135 ISSN: 0168-1656 ISO Abbreviation: J. Biotechnol. Publication Date: 2008 Jun |
Date Detail:
|
Created Date: 2008-06-23 Completed Date: 2008-10-06 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 8411927 Medline TA: J Biotechnol Country: Netherlands |
Other Details:
|
Languages: eng Pagination: 272-80 Citation Subset: IM |
Affiliation:
|
Biotchnology Research Institute, 6100 Royalmount Avenue, Montreal, Quebec, Canada. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Alkaline Phosphatase
/
metabolism* Analysis of Variance Animals Baculoviridae / metabolism* Bioreactors* CHO Cells Cell Line Cricetinae Cricetulus Filtration Genetic Vectors Humans Insects Transduction, Genetic* |
| Chemical | |
Reg. No./Substance:
|
EC 3.1.3.1/Alkaline Phosphatase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Investigating the fermentation of cocoa by correlating denaturing gradient gel electrophoresis profi...
Next Document: Rationalizing the selection of oral lipid based drug delivery systems by an in vitro dynamic lipolys...