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Hydrazide‐Functionalized Poly(2‐hydroxyethyl methacrylate) Microspheres for Immobilization of Horseradish Peroxidase
Author(s) -
Horák Daniel,
Karpíšek Michal,
Turková Jaroslava,
Beneš Milan
Publication year - 1999
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp990006w
Subject(s) - horseradish peroxidase , chemistry , methacrylate , polymerization , polymer chemistry , dispersion polymerization , suspension polymerization , (hydroxyethyl)methacrylate , nuclear chemistry , organic chemistry , polymer , enzyme
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 μg 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.