| A Hydrogel is defined as a polymeric material which possesses the ability to swell in water and retain a significant fraction of water within its structure, but which will not dissolve in water. Hydrogels have been studied by many researchers because they have many useful applications in bio related fields such as drug delivery, bioseparation, and etc.; In this thesis, a new hydrogel system that possesses the characteristics of thermo-responsive swelling property and immobilized metal affinity was developed. This affinity material consists of a hydrogel with stimuli responsive swelling characteristics to provide modulated diffusivity and size selectivity. Covalently bound ligands within hydrogels provide highly selective and tunable affinity-based separation. Swelling and affinity properties can be independently controlled by regulating the temperature or pH of the solution to provide a sequential separations scheme. The developed affinity hydrogels incorporate multiple modes of separations or recovery and concentrate specific solutes in chromatographic systems.; Thermal sensitive affinity hydrogels were synthesized from a N-isopropylacrylamide (NIPAAm) monomer, a crosslinker (1,4-bismethylene acrylamide) and a ligand attachable co-monomer acrylamide (AAm), using free radical chemistry. The ligand of choice is the metal affinity iminodiacetic acid (IDA) which is bound to hydrogel backbone via a spacer arm. The challenge lay in incorporating affinity ligands without affecting the temperature induced swelling of the hydrogel. Thus, PNIPAAm-Am hydrogels are functionalized with a spacer arm (1,4-butanediol diglycidyl ether), the chelating ligand IDA and a divalent metal ion (Cu2+). This ligand binds histidine groups at high pH and releases them upon protonation of histidine at low pH. This can be used to separate proteins based on the occurrence of surface histidine residues in them. The resulting affinity hydrogel was shown to adsorb the protein chicken egg white lysozyme at high pH and desorbs it at low pH. The number of functional groups within a hydrogel was measured by bound copper content (BCC) method. The hydrogel's thermal swelling properties, lysozyme adsorption and desorption phenomena were also measured. |
