Hydrazide-functionalized poly(2-hydroxyethyl methacrylate) microspheres for immobilization of horseradish peroxidase.
Nonporous cross-linked poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (poly(HEMA-co-EDMA)) microspheres were prepared by dispersion polymerization of HEMA and EDMA. The polymerization was performed in toluene/2-methylpropan-1-ol in the presence of cellulose acetate butyrate as a steric stabilizer and dibenzoyl peroxide initiator. The particle size may be increased by decreasing the toluene/2-methylpropan-1-ol ratio and by increasing polymerization temperature. Adipohydrazide was attached to the microspheres activated with 2,4,6-trichloro-1,3,5-triazine. After periodate oxidation of its carbohydrate moieties, horseradish peroxidase was coupled to the hydrazide-functionalized poly(HEMA-co-EDMA) microparticles up to 7.3 microgram of enzyme/g of carrier without a significant loss of its activity. Immobilized peroxidase was found to be stable, retaining more than 97% of its initial activity when stored for 23 days after the preparation.
Horák
D
D
Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovský Sq. 2, 162 06 Prague 6, Czech Republic.
Karpísek
M
M
Turková
J
J
Benes
M
M
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000378:metabolism; D008689:Methacrylates / Q000737:chemistry; D008863:Microspheres; D009584:Nitrogen / Q000737:chemistry; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry*
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D008689:Methacrylates; D011102:Polyhydroxyethyl Methacrylate; C004919:ethylene dimethacrylate; D006735:Horseradish Peroxidase; D009584:Nitrogen