Abstract
Magnetic nanoparticles, covered by a polymeric hydrophilic nanolayer containing reactive amino groups, were obtained via Hoffman degradation of the polyacrylamide-coated Fe 3O 4 nanoparticles synthesized by photochemical in situ polymerization, and then conjugated the model enzyme––α-chymotrypsin (CT) by use of EDC· HCl and NHS at room temperatures. The mechanism of photochemical in situ polymerization was briefly proposed in this paper. Superparamagnetic properties were retained for Fe 3O 4 after enzyme immobilization while slightly reducing the value of saturation magnetization. Crystalline structure of Fe 3O 4 after CT immobilization was consistent with that of the freshly prepared Fe 3O 4 by X-ray diffraction (XRD) analysis. The binding capacity was 69 and 61 mg enzyme/g nanogel determined by thermogravimetric (TG) analysis and by standard BCA protein assay, respectively. Specific activity of the immobilized CT was 0.93 U/(mg min), only 59.3% as that of free CT. Thermal stability of CT was improved after being bound to the amine-functionalized magnetic nanogel.
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