A polyacrylonitrile (PAN)-based immobilized metal-ion affinity membrane (IMAM) was prepared with a high capacity for protein adsorption. PAN was selected as the substrate due to its excellent thermal and chemical stability. The cyano groups on the PAN membrane were substituted with carboxyl groups, followed by reactions with ethylenediamine (EDA) and ethylene glycol diglycidyl ether (EGDGE) to produce the terminal epoxy groups. The chelating agent iminodiacetic acid (IDA) was then bound to the modified PAN membrane and further chelated with copper ions. The immobilized copper ion amount of membrane was analyzed to obtain the optimal reaction conditions, which were 60 °C/3 h for EDA coupling and 60 °C/4 h for EGDGE grafting. Furthermore, under the use of minor IDA and copper ion concentrations, the immobilized copper ion capacity of the IMAM was 4.8 μmol/cm2 (253.4 µmol/mL, or 1.47 μmol/mg). At a neutral pH, the cationic lysozyme exhibited a large adsorption capacity with the IMAM (1.96 μmol/mL), which was most likely multilayer binding, whereas the adsorption capacity for bovine serum albumin (BSA) and histidine-tagged green fluorescent protein (GFP-His6) was 0.053 μmol/mL and 0.135 μmol/mL, respectively, with a monolayer adsorption arrangement. The protein desorption efficiency was greater than 95%, implying that the prepared IMAM could be reused for protein adsorption.
Keywords: adsorption arrangement; immobilized metal-ion affinity membrane; polyacrylonitrile membrane; protein adsorption.