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Properties of RNase A Immobilized on Magnetic Poly(2-hydroxyethyl methacrylate) Microspheres

Authors

  • Daniel Horák,

    Corresponding author
    1. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06 Prague 6, Czech Republic
    • Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06 Prague 6, Czech Republic
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  • Bohuslav Rittich,

    1. Masaryk University in Brno, Faculty of Science, Institute of Microbiology, Tvrdého 14, 602 00 Brno, Czech Republic
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  • Jan Šafář,

    1. Masaryk University in Brno, Faculty of Science, Institute of Microbiology, Tvrdého 14, 602 00 Brno, Czech Republic
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  • Alena Španová,

    1. Masaryk University in Brno, Faculty of Science, Institute of Microbiology, Tvrdého 14, 602 00 Brno, Czech Republic
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  • Jiří Lenfeld,

    1. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06 Prague 6, Czech Republic
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  • Milan J. Beneš

    1. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám. 2, 162 06 Prague 6, Czech Republic
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Abstract

Magnetic hydrogel microspheres 1.5 μm in size were prepared by dispersion copolymerization of 2-hydroxyethyl methacrylate and ethylene dimethacrylate in the presence of magnetite, which formed the core of the particles. RNase A was coupled to the particles by the cyanuric chloride method. Gel electrophoresis of plasmid DNA pUC 19 (contaminated by bacterial RNA) confirmed RNA degradation with the immobilized enzyme. The effect of temperature and pH on the relative activity of immobilized RNase A was estimated after incubation of the samples at different temperatures (30−80 °C) and pH (4.0−8.0). Maximum relative activity was observed at 70 °C and pH 6.5. The matrices based on magnetic poly(HEMA) had a low tendency to adsorb RNA.

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