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Degradation of supercoiled plasmid DNA within a capillary device

Authors

  • F.J. Meacle,

    1. The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
    2. Merck Research Labs, Westpoint, Pennsylvania
    Current affiliation:
    1. Centocor R&D, 145 King of Prussia Road, Radner, Pennsylvania 19087
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  • H. Zhang,

    1. The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
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  • I. Papantoniou,

    1. The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
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  • J.M. Ward,

    1. Department of Biochemistry and Molecular Biology, University College London, Gower Street, WC1E 6BT, United Kingdom
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  • N.J. Titchener-Hooker,

    1. The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
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  • M. Hoare

    Corresponding author
    1. The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
    • The Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, United Kingdom; telephone: 0044-2076797031; fax: 0044-2072090703
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  • Both FJM and HZ made an equal contribution.

Abstract

Supercoiled plasmid DNA is susceptible to fluid stress in large-scale manufacturing processes. A capillary device was used to generate controlled shear conditions and the effects of different stresses on plasmid DNA structure were investigated. Computational fluid dynamics (CFD) analysis was employed to characterize the flow environment in the capillary device and different analytical techniques were used to quantify the DNA breakage. It was found that the degradation of plasmid DNA occurred at the entrance of the capillary and that the shear stress within the capillary did not affect the DNA structure. The degradation rate of plasmids was well correlated with the average elongational strain rate or the pressure drop at the entrance region. The conclusion may also be drawn that laminar shear stress does not play a significant role in plasmid DNA degradation. Biotechnol. Bioeng. 2007; 97: 1148–1157. © 2006 Wiley Periodicals, Inc.

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