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

Citation
D. Horak et al., Properties of RNase a immobilized on magnetic poly(2-hydroxyethyl methacrylate) microspheres, BIOTECH PR, 17(3), 2001, pp. 447-452
Citations number
33
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY PROGRESS
ISSN journal
87567938 → ACNP
Volume
17
Issue
3
Year of publication
2001
Pages
447 - 452
Database
ISI
SICI code
8756-7938(200105/06)17:3<447:PORAIO>2.0.ZU;2-F
Abstract
Magnetic hydrogel microspheres 1.5 mum 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. RNas e A was coupled to the particles by the cyanuric chloride method. Gel elect rophoresis 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 incu bation of the samples at different temperatures (30-80 degreesC) and pH (4. 0-8.0). Maximum relative activity was observed at 70 degreesC and pH 6.5. T he matrices based on magnetic poly(HEMA) had a low tendency to adsorb RNA.