Cryogels containing imidazole functional group for lysozyme adsorption

In this thesis, super macroporous cryogels were synthesized and used to purify lysozyme from an aqueous solution. Imidazole functional groups were incorporated into a basic polymeric skeleton to impart pseudo-specificity to the cryogels using n-vinylimidazole as the comonomer. Cryogels were prepared by cryogelation in an aqueous solution of partially frozen monomers in a plastic syringe column. polyacrylamide-N-vinyl imidazole) [poly(Aam-VIM)] cryogels were prepared by polymerization of N-vinyl imidazole, a water-soluble and functional monomer as affinity ligand, with Acrylamide monomer for hydrophilic and gelling purposes in the presence of N-N Methylenebisacrylamide as cross-linker. Within the scope of characterization studies to evaluate the physical and chemical properties of the synthesized cryogels, swelling test, Fourier transform infrared spectroscopy (FTIR), elemental analysis, Field Emission Scanning Electron Microscope (FESEM), and Thermo Gravimetric Analysis (TGA-DTA) methods were performed. Lysozyme adsorption studies from aqueous solution with poly(AAm-VIM) cryogels were carried out in pH 9 (TRIS buffer). In order to determine the optimum conditions in the adsorption process, the most suitable conditions for lysozyme adsorption were determined in the light of parameters such as initial concentration screening, ionic strength effect, temperature effect, and flow rate of the column. Langmuir and Freundlich's isotherm models were studied as isotherm models to evaluate the adsorption parameters mathematically. According to the regression coefficient results obtained, it was determined that lysozyme adsorption was more compatible with the Langmuir isotherm model. As a result of SDS-PAGE analysis, purity control of the pol(AAm-VIM) cryogel column was carried out to separate lysozyme from the aqueous solution. The inertness and regeneration ability of poly(AAm-VIM) cryogel affinity columns were measured with reusability experiments performed within the adsorption-desorption cycle.