Document Detail


Enhanced CHO Cell-Based Transient Gene Expression with the Epi-CHO Expression System.
MedLine Citation:
PMID:  21104043     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Transient gene expression systems in mammalian cells continue to grow in popularity due to their capacity to produce significant amounts of recombinant protein in a rapid and scalable manner, without the lengthy time periods and resources required for stable cell line development. Traditionally, production of recombinant monoclonal antibodies for pre-clinical assessment by transient expression in CHO cells has been hampered by low titers. In this report, we demonstrate transient monoclonal antibody titers of 140 mg/l with CHO cells using the episomal-based transient expression system, Epi-CHO. Such titers were achieved by implementing an optimized transfection protocol incorporating mild-hypothermia and through screening of a variety of chemically defined and serum-free media for their ability to support elevated and prolonged viable cell densities post-transfection, and in turn, improve recombinant protein yields. Further evidence supporting Epi-CHO's capacity to enhance transgene expression is provided, where we demonstrate higher transgene mRNA and protein levels of two monoclonal antibodies and a destabilized enhanced green fluorescent protein with Epi-CHO compared to cell lines deficient in plasmid DNA replication and/or retention post-transfection. The results demonstrate the Epi-CHO system's capacity for the rapid production of CHO cell-derived recombinant monoclonal antibodies in serum-free conditions.
Authors:
Joe Codamo; Trent P Munro; Benjamin S Hughes; Michael Song; Peter P Gray
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Molecular biotechnology     Volume:  48     ISSN:  1559-0305     ISO Abbreviation:  Mol. Biotechnol.     Publication Date:  2011 Jun 
Date Detail:
Created Date:  2011-04-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9423533     Medline TA:  Mol Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  109-15     Citation Subset:  IM    
Affiliation:
The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD, 4072, Australia, g.codamo@uq.edu.au.
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