Enhanced CHO cell-based transient gene expression with the Epi-CHO expression system

Codamo, Joe, Munro, Trent P., Hughes, Benjamin S., Song, Michael and Gray, Peter P. (2011) Enhanced CHO cell-based transient gene expression with the Epi-CHO expression system. Molecular Biotechnology, 48 2: 109-115. doi:10.1007/s12033-010-9351-9


Author Codamo, Joe
Munro, Trent P.
Hughes, Benjamin S.
Song, Michael
Gray, Peter P.
Title Enhanced CHO cell-based transient gene expression with the Epi-CHO expression system
Formatted title
Enhanced CHO cell-based transient gene expression with the Epi-CHO expression system
Journal name Molecular Biotechnology   Check publisher's open access policy
ISSN 1073-6085
1559-0305
Publication date 2011-06-01
Year available 2010
Sub-type Article (original research)
DOI 10.1007/s12033-010-9351-9
Open Access Status Not yet assessed
Volume 48
Issue 2
Start page 109
End page 115
Total pages 7
Place of publication Totowa, NJ, United States
Publisher Humana Press
Language eng
Abstract 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.
Formatted abstract
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.
Keyword Transient gene expression
CHO
Monoclonal antibodies
Episomal
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published online: 20 November 2010

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 17 times in Thomson Reuters Web of Science Article | Citations
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Created: Sat, 01 Oct 2011, 01:04:50 EST by Mr Benjamin Hughes on behalf of Aust Institute for Bioengineering & Nanotechnology