Publication details

Properties of RNase A immobilized on magnetic poly(2-hydroxyethyl methacrylate) microspheres

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Authors

HORÁK Daniel RITTICH Bohuslav ŠAFÁŘ Jan ŠPANOVÁ Alena LENFELD Jiří BENEŠ Milan J.

Year of publication 2001
Type Article in Periodical
Magazine / Source Biotechnology Progress
MU Faculty or unit

Faculty of Science

Citation
Field Biotechnology
Keywords Dispersion polymerization; RNase A; nucleic acids; particles; polystyrene; chromatography; purification
Description Magnetic hydrogel microsheres were prepared by dispersion copolymerisation 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 pUC19 (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 matrice based on magnetic poly(HEMA) had a low tendency to adsorb RNA.
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