Preparation of Thermo-Sensitive Magnetic Hydrogel Microspheres and Application to Enzyme Immobilization
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概要
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Magnetic hydrogel microspheres were parepared by co-polymerizing N-isopropylacrylamide (NIPAM), methacrylic acid (MAA) and N, N'-methylene-bis-acrylamide (MBA) in the presence of ultrafine magnetite particles (average particle diameter of 10-20 nm) prepared by co-precipitation. The synthesized magnetic hydrogel microspheres with varying MAA content were monodispersed, and their average diameters ranged from 150-250 nm. All the magnetic hydrogel microspheres showed a reversible transition between flocculation and dispersion as a function of temperature (4-40℃), and the thermo-flocculated microspheres were separated quickly in the magnetic field. Enzymes, were immobilized onto these thermo-sensitive magnetic hydrogel microspheres by two different methods. Trypsin was covalently immobilized onto the magnetic hydrogel microspheres in high yield by the carbodiimide method. In the case of micropheres with higher MAA content, immobilized trypsin demonstrated higher activity. On the other hand, a fuasion enzyme constituing of affinity tag AG (immunoglobulin G binding domains) and β-galactosidase (βgal) was immobilized onto the magnetic hydrogel microspheres with covalenity immobilized human γ-globulin by affinity interaction. The immobilized AGβgal retained high activity even in the case of microspheres with a low MAA content. Therefore, the thermosensitive magnetic hydrogel microspheres are useful carries for enzyme immobilization.
- 公益社団法人日本生物工学会の論文
- 1997-10-25
著者
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Fukuda Hideki
Division Of Molecular Science Graduate School Of Science And Technology Kobe University
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Kondo Akihiko
Department Of Applied Chemistry Kyushu Institute Of Technology
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Kondo Akihiko
Department Of Applied Chemistry Faculty Of Engineering Kyushu Institute Of Technology
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Fukuda Hideki
Division Of Molecular Science Graduate Schol Of Science And Technology Kobe University
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Fukuda Hideki
Division Of Bioscience The Graduate School Of Science And Technology Kobe University
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