Precipitation of filamentous bacteriophages for their selective recovery in primary purification

Authors

  • Steven Branston,

    1. The Advanced Centre for Biochemical Engineering, Dept. of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, U.K
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  • Emma Stanley,

    1. Institute of Structural and Molecular Biology, Div. of Biosciences, University College London, Gower Street, London WC1E 6BT, U.K
    Current affiliation:
    1. Novartis Pharmaceuticals UK Limited, Horsham Research Centre, Wimblehurst Road, Horsham, West Sussex, RH12 5AB, U.K
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  • Eli Keshavarz-Moore,

    Corresponding author
    1. The Advanced Centre for Biochemical Engineering, Dept. of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, U.K
    • The Advanced Centre for Biochemical Engineering, Dept. of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, U.K
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  • John Ward

    1. Institute of Structural and Molecular Biology, Div. of Biosciences, University College London, Gower Street, London WC1E 6BT, U.K
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Abstract

Filamentous bacteriophages and their derivatives are showing great promise as a whole new class of industrial agents, such as biologically based nano-materials and viral vectors. This raises challenges for their large-scale manufacture, principally due to the lack of bioprocessing knowledge. This article addresses what will be a potentially important option in the primary purification of the bacteriophages. Polyethylene glycol (PEG)-salt dual precipitants, calcium ions, spermidine, and isoelectric precipitation were first examined for their potential suitability for bacteriophage concentration under both pure and broth conditions. Successful precipitants were further studied on the basis of their selective purification ability from DNA and protein contaminants in a clarified broth system. Both PEG-based and isoelectric precipitations resulted in bacteriophage purity improvements, and PEG-based precipitations offered the highest selectivities. This work shows that precipitation of bacteriophages can be an effective primary purification step in a large-scale bioprocess. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012

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