Cytochrome c adsorption by supermacroporous monolithic phema affinity cryogel membranes embedded with Cu2+ attached sporopollenin particles

In this study, adsorption of cytochrome c onto supermacroporous monolithic PHEMA affinity cryogels embedded with Cu2+ - attached sporopollenin was investigated. For this purpose, firstly, sporopollenin particles, with size of 20 ? m, were treated with 150 ppm Cu2+ solution at pH 5,0 to bond Cu2+ ions to the structure. Then, PHEMA based monolithic affinity cryogels containing 15 mg of embedded Cu2+- sporopollenin were synthesized by free radical polymerization using, 2-hydroxyethyl methacrylate (HEMA) as a monomer and N,N´-methylene-bis-acrylamide (MBAAm) as a cross-linker in a plastic syringe of 5 mL. Monolithic affinity cryogel sorbents embedded with Cu2+- sporopollenin were used in adsorption studies by changing the conditions (pH, initial cytochrome c concentration, temperature, flow rate and ionic strength). The prepared cryogels were characterized by swelling tests and scanning electron microscopy (SEM). The maximum amount of cytochrome c adsorption from aqueous solution was observed at pH 7,0. In concentration studies, the equilibrium was established about 3,0 mg/mL of cytochrome c concentration. In addition, it was observed that cytochrome c adsorption was inversely proportional to flow rate and temperature. It was also observed that non-specific adsorption of cytochrome c onto affinity cryogel membranes containing sporopollenin particles, without atteched Cu2+, was ignoreable degree. Adsorption/desorption processes were applied 30 times with these affinity cryogel sorbents without noticeable loss in their adsorption capacity.Keywords: Cryogel, PHEMA, Sporopollenin, Cytochrome c, IMAC, Adsorption.