M. Nalan TÜZMEN, Nilay BERELİ, Müge ANDAÇ, Sinan AKGÖL, Adil DENİZLİ

Supermacroporous Support Having Dye for Reuseable Enzyme Immobilization

Poly acrylamide-allyl glycidyl ether [poly AAm-AGE ] cryogel was prepared by bulk polymerization which proceedsin an aqueous solution of monomers frozen inside a glass column cryo-polymerization . After thawing, the monolithiccryogel contains a continuous polymeric matrix having interconnected pores of 10-100 µm size. Cibacron Blue F3GAwas attached by covalent binding onto poly AAm-AGE cryogel via epoxy groups. Poly AAm-AGE cryogel wascharacterized by FTIR and scanning electron microscopy SEM . The equilibrium swelling degree of the poly AAmAGE monolithic cryogel was 6.84 g H2O/g cryogel. Poly AAm-AGE cryogel containing 68.9 µmol Cibacron BlueF3GA/g was used in the adsorption ofβ-casein from aqueous solution. The effects of initial concentration, pH, ionicstrength and flow rate on the adsorption efficiency of cryogel were studied in a column system. The non-specificadsorption ofβ-casein was very low 1.26 mg/g polymer . The maximum amount of β-casein adsorption from aqueoussolution in acetate buffer was 44.7 mg/g at pH 4.0. Up to 99% of the adsorbedβ-casein was eluted when 0.1 MTris/HCl buffer containing 0.5 M NaCl was used as elution agent. We concluded that Cibacron Blue F3GA attachedpoly AAm-AGE cryogel could be repeatedly applied forβ-casein adsorption without significant losses in the adsorptioncapacity

Keywords:

β-Casein, cryogels, Dye-affinity Chromatography, Protein Purification,

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