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    Cultivation of E. coli carrying a plasmid-based Measles vaccine construct (4.2 kbp pcDNA3F) employing medium optimisation and pH-temperature induction techniques

    Access Status
    Open access via publisher
    Authors
    Ongkudon, C.
    Pickering, R.
    Webster, D.
    Danquah, Michael
    Date
    2011
    Type
    Journal Article
    
    Metadata
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    Citation
    Ongkudon, C. and Pickering, R. and Webster, D. and Danquah, M. 2011. Cultivation of E. coli carrying a plasmid-based Measles vaccine construct (4.2 kbp pcDNA3F) employing medium optimisation and pH-temperature induction techniques. Microbial Cell Factories. 10: Article 16.
    Source Title
    Microbial Cell Factories
    DOI
    10.1186/1475-2859-10-16
    School
    Curtin Sarawak
    URI
    http://hdl.handle.net/20.500.11937/13753
    Collection
    • Curtin Research Publications
    Abstract

    Background: Plasmid-based measles vaccines offer great promises over the conventional fertilised egg method such as ease of manufacture and mimic wild-type intracellular antigen expression. The increasing number of clinical trials on plasmid-based measles vaccines has triggered the need to make more in less time. Results: In this work, we investigated the process variables necessary to improve the volumetric and specific yields of a model plasmid-based measles vaccine (pcDNA3F) harboured in E. coli DH5α. Results from growth medium optimisation in 500 mL shake flasks by response surface methodology (RSM) generated a maximum volumetric yield of 13.65 mg/L which was 1.75 folds higher than that of the base medium. A controlled fed-batch fermentation employing strategic glycerol feeding and optimised growth conditions resulted in a remarkable pcDNA3F volumetric yield of 110 mg/L and a specific yield of 14 mg/g. In addition, growth pH modification and temperature fluctuation between 35 and 45°C were successfully employed to improve plasmid production. Conclusion: Production of a high copy number plasmid DNA containing a foreign gene of interest is often hampered by the low plasmid volumetric yield which results from the over expression of foreign proteins and metabolic repressors. In this work, a simple bioprocess framework was employed and successfully improved the production of pcDNA3F.

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