| Enhanced CHO Cell-Based Transient Gene Expression with the Epi-CHO Expression System. | |
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MedLine Citation:
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PMID: 21104043 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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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. |
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Authors:
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Joe Codamo; Trent P Munro; Benjamin S Hughes; Michael Song; Peter P Gray |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Molecular biotechnology Volume: 48 ISSN: 1559-0305 ISO Abbreviation: Mol. Biotechnol. Publication Date: 2011 Jun |
Date Detail:
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Created Date: 2011-04-22 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9423533 Medline TA: Mol Biotechnol Country: United States |
Other Details:
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Languages: eng Pagination: 109-15 Citation Subset: IM |
Affiliation:
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The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, Brisbane, QLD, 4072, Australia, g.codamo@uq.edu.au. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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