Thermosensitive molecularly imprinted hydrogel cross-linked with N-malely chitosan for the recognition and separation of BSA

A novel temperature-sensitive molecularly imprinted hydrogel composed of N-isopropylacrylamide and acrylamide has been prepared by using free-radical polymerization and was cross-linked by modified water-soluble N-maley chitosan in aqueous solution. BSA (pI 4.9, MW 66.0 kDa) was used as the template protein. The produced hydrogels were characterized by environmental SEM to reveal the microcosmic morphology. A microporous structure was only found in the imprinted hydrogel, while no obvious microporous structure was found in nonimprinted hydrogels. The lower critical solution temperature of the hydrogels was 34 degrees C, and the optimal binding conditions were tested, namely, the adsorption equilibrium time of 6 h and initial BSA concentration of 1.0 mg/mL. The adsorption capacity Q(max) was determined by Langmuir isotherm plots and was 5.72 mg/g for imprinted hydrogel and 1.18 mg/g for nonimprinted hydrogels. A separation factor () of 4 was obtained when bovine hemoglobin (pI 6.9, MW 64.0 kDa) was selected as the particular reference protein. Molecular weights and pIs were chosen to investigate the selectivity of the hydrogels. It was shown that the shape memory and the size effect were the major factors for the recognition. This imprinted hydrogel was used to specifically adsorb the BSA from the protein mixture.