Preparation and characterization of magnetic nanoparticles for urokinase purification

In this thesis, magnetic poly(2-hydroxyethyl methacrylate) [p(HEMA)] nanoparticles were synthesized by surfactant free emulsion polymerization technique. Cibacron Blue F3GA was covalently attached to the magnetic p(HEMA) nanoparticles and Cu2+ ions were then chelated to dye molecules. Urokinase was adsorbed onto Cu2+ -Cibacron Blue F3GA derived magnetic p(HEMA) nanoparticles. Synthesized magnetic nanoparticles was about 80 nm in diameter, spherical and carried magnetic character. Incorporation rate of Cibacron Blue for magnetic nanoparticles was found to be 28.125 µmol/g polymer. The Cu2+ ions loaded was calculated to be 10.229 µmol/g polymer. This Cu2+ -Cibacron Blue F3GA derived magnetic p(HEMA) nanoparticles were used for urokinase adsorption under different conditions (i.e. pH, enzyme initial concentration, ionic strength, temperature). Maximum adsorption capacity was found to be 630.43 mg/g polymer for Cu2+-Cibacron Blue F3GA derived magnetic p(HEMA) nanoparticles. It was observed that Langmuir adsorption isotherm was applicable in this adsorption process. The urokinase adsorbed Cu2+-Cibacron Blue F3GA derived magnetic p(HEMA) nanoparticles was desorbed with 1.0 M NaCl and desorption rate was found to be 96 %. It was also analyzed that adsorption capacity did not changed significantly after five adsorption/desorption cycles. This study promised that synthesized support material could be used for urokinase purification from biological samples.