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    Large-volume methacrylate monolith for plasmid purification. Process engineering approach to synthesis and application

    Access Status
    Fulltext not available
    Authors
    Danquah, Michael
    Forde, G.
    Date
    2008
    Type
    Journal Article
    
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    Citation
    Danquah, M. and Forde, G. 2008. Large-volume methacrylate monolith for plasmid purification. Process engineering approach to synthesis and application. Journal of Chromatography A. 1188 (2): pp. 227-233.
    Source Title
    Journal of Chromatography A
    DOI
    10.1016/j.chroma.2008.02.045
    ISSN
    0021-9673
    School
    Curtin Sarawak
    URI
    http://hdl.handle.net/20.500.11937/34063
    Collection
    • Curtin Research Publications
    Abstract

    The extent of exothermicity associated with the construction of large-volume methacrylate monolithic columns has somewhat obstructed the realisation of large-scale rapid biomolecule purification especially for plasmid-based products which have proven to herald future trends in biotechnology. A novel synthesis technique via a heat expulsion mechanism was employed to prepare a 40 mL methacrylate monolith with a homogeneous radial pore structure along its thickness. Radial temperature gradient was recorded to be only 1.8 °C. Maximum radial temperature recorded at the centre of the monolith was 62.3 °C, which was only 2.3 °C higher than the actual polymerisation temperature. Pore characterisation of the monolithic polymer showed unimodal pore size distributions at different radial positions with an identical modal pore size of 400 nm. Chromatographic characterisation of the polymer after functionalisation with amino groups displayed a persistent dynamic binding capacity of 15.5 mg of plasmid DNA/mL. The maximum pressure drop recorded was only 0.12 MPa at a flow rate of 10 mL/min. The polymer demonstrated rapid separation ability by fractionating Escherichia coli DH5a-pUC19 clarified lysate in only 3 min after loading. The plasmid sample collected after the fast purification process was tested to be a homogeneous supercoiled plasmid with DNA electrophoresis and restriction analysis. © 2008 Elsevier B.V. All rights reserved.

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