Preparation of ionic activation of cryogel monolith columns for purification of poly-L-lysine

Cryogels are large-pored, high-water-holding polymeric structures produced at sub-zero temperatures. These polymers are used in many fields including bioprocess. The aim of this thesis is primarily to produce and use cryogel monolith columns for the separation and purification of Ɛ-poly-L-lysine molecules. For this reason, cryogel columns were prepared using acrylamide, hydroxyethyl acrylate and hydroxyethyl methacrylate at different temperatures and monomer amounts and catalysts. Afterwards, they were activated with cationic and anionic loadings and their characterizations were made. In these characterizations, scanning electron microscope imaging was performed, swelling capacity and porosity, ion exchange charge, degree of cross-linking were calculated, and chemical structure characterization was performed. In order to understand the binding capacities, static binding and dynamic binding experiments were performed. Sodium chloroacetate was used for anionic loading and iodomethane was used for cationic loading. Among the columns selected in the studies, the Ɛ-PL binding capacity of anionic loaded acrylamide columns was found to be between 0.009 g/mg and 1.691 g/mg. The binding capacities of the cationic loaded hydroxyethyl methacrylate columns for bovine serum albumin were found to be 0.214 g/mg and 1.132 g/mg. Although the binding capacity of the columns is low, they can be used at high flow rates. However, it has been understood as a result of the experiments that the produced columns are not suitable for continuous chromatography and that they are disposable. As a result, more studies are needed on the use of cryogel columns in bioprocesses.