The production of phema microbead embedded supermacroporous cryogel composit membranes and investigation of their protein adsorption capacities

In this study, Immobilized Metal Affinity Chromatograph (IMAC) finding a wide application area among affinity chromatography methods was used for removal of lysozyme efficiently from aqueous solutions. Poly(2-hydroxyethyl methacrylate) beads with the size of about 2 µm were prepared, and than iminodiacetic acid (IDA) was covalently attached to these beads. Then, Cu2+ ions were attached to PHEMA beads modified by IDA. And finally, supermacroporous poly (2-hydroxyethyl methacrylate) base monolithic cryogel column embedded with PHEMA-Cu2+ beads. For this purpose, supermacroporous composite cryogels were prepared by free radical polymerization using N,N,N´,N´-tetramethylene diamine (TEMED) and ammonium persulfate (APS) as initiator/activator pair, N,N´-methylene-bis-acrylamide (MBAAm) as cross-linker in a plastic syringe of 5 mL. Prepared supermacroporous cryogels with embedded PHEMA-Cu2+ beads were characterized by FTIR, SEM, sweling tests and surface area measurements. The effect of pH, buffer, ionic strength, initial lysozyme concentration, temparature and flow rate on adsorption were investigated. The maximum amount of lysozyme adsorption from aqueous solution was at 107,14 mg/g polymer in Tris-Base buffer at pH 9 with initial lysozyme concentration of 5 mg/mL. It was also observed that enzyme could be repeatedly adsorbed and desorbed on the supermacroporous cryogels with embedded PHEMA-Cu2+ beads.