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Rational engineering of Escherichia coli strains for plasmid biopharmaceutical manufacturing

Authors

  • Geisa A. L. Gonçalves,

    1. Department of Bioengineering, Instituto Superior Técnico (IST), Lisbon, Portugal
    2. Center for Biological and Chemical Engineering, IBB-Institute for Biotechnology and Bioengineering, IST, Lisbon, Portugal
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    • These authors contributed equally to this work.

  • Diana M. Bower,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
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    • These authors contributed equally to this work.

  • Duarte M. F. Prazeres,

    1. Department of Bioengineering, Instituto Superior Técnico (IST), Lisbon, Portugal
    2. Center for Biological and Chemical Engineering, IBB-Institute for Biotechnology and Bioengineering, IST, Lisbon, Portugal
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  • Gabriel A. Monteiro,

    1. Department of Bioengineering, Instituto Superior Técnico (IST), Lisbon, Portugal
    2. Center for Biological and Chemical Engineering, IBB-Institute for Biotechnology and Bioengineering, IST, Lisbon, Portugal
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  • Dr. Kristala L. J. Prather

    Corresponding author
    1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
    • Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., Room 66-454, Cambridge, MA 02139, USA
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Abstract

Plasmid DNA (pDNA) has become very attractive as a biopharmaceutical, especially for gene therapy and DNA vaccination. Currently, there are a few products licensed for veterinary applications and numerous plasmids in clinical trials for use in humans. Recent work in both academia and industry demonstrates a need for technological and economical improvement in pDNA manufacturing. Significant progress has been achieved in plasmid design and downstream processing, but there is still a demand for improved production strains. This review focuses on engineering of Escherichia coli strains for plasmid DNA production, understanding the differences between the traditional use of pDNA for recombinant protein production and its role as a biopharmaceutical. We will present recent developments in engineering of E. coli strains, highlight essential genes for improvement of pDNA yield and quality, and analyze the impact of various process strategies on gene expression in pDNA production strains.

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