Supermacroporous support having dye for reuseable enzyme immobilization

Abstract

Poly(acrylamide-allyl glycidyl ether) [poly(AAm-AGE)] cryogel was prepared by bulk polymerization which proceeds in an aqueous solution of monomers frozen inside a glass column (cryo-polymerization). After thawing, the monolithic cryogel contains a continuous polymeric matrix having interconnected pores of 10-100 μm size. Cibacron Blue F3GA was attached by covalent binding onto poly(AAm-AGE) cryogel via epoxy groups. Poly(AAm-AGE) cryogel was characterized by FTIR and scanning electron microscopy (SEM). The equilibrium swelling degree of the poly(AAm- AGE) monolithic cryogel was 6.84 g H2O/g cryogel. Poly(AAm-AGE) cryogel containing 68.9 μmol Cibacron Blue F3GA/g was used in the adsorption of β-casein from aqueous solution. The effects of initial concentration, pH, ionic strength and flow rate on the adsorption efficiency of cryogel were studied in a column system. The non-specific adsorption of β-casein was very low (1.26 mg/g polymer). The maximum amount of β-casein adsorption from aqueous solution in acetate buffer was 44.7 mg/g at pH 4.0. Up to 99% of the adsorbed β-casein was eluted when 0.1 M Tris/HCl buffer containing 0.5 M NaCl was used as elution agent. We concluded that Cibacron Blue F3GA attached poly(AAm-AGE) cryogel could be repeatedly applied for β-casein adsorption without significant losses in the adsorption capacity.