Yang Yin-Jie, Chang Hou-Chien, Wang Min-Ying, Suen Shing-Yi
Department of Chemical Engineering, National Chung Hsing University, Taichung 402, Taiwan.
Graduate Institute of Biotechnology, National Chung Hsing University, Taichung 402, Taiwan.
Membranes (Basel). 2023 Feb 24;13(3):271. doi: 10.3390/membranes13030271.
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/cm (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-His) 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.
制备了一种基于聚丙烯腈（PAN）的固定化金属离子亲和膜（IMAM），其具有高蛋白质吸附容量。由于PAN具有优异的热稳定性和化学稳定性，因此被选作基质。PAN膜上的氰基被羧基取代，随后与乙二胺（EDA）和乙二醇二缩水甘油醚（EGDGE）反应生成末端环氧基。然后将螯合剂亚氨基二乙酸（IDA）结合到改性PAN膜上，并进一步与铜离子螯合。分析膜的固定化铜离子量以获得最佳反应条件，即EDA偶联为60℃/3小时，EGDGE接枝为60℃/4小时。此外，在使用少量IDA和铜离子浓度的情况下，IMAM的固定化铜离子容量为4.8μmol/cm（253.4μmol/mL，或1.47μmol/mg）。在中性pH值下，阳离子溶菌酶对IMAM表现出较大的吸附容量（1.96μmol/mL），这很可能是多层结合，而牛血清白蛋白（BSA）和组氨酸标签绿色荧光蛋白（GFP-His）的吸附容量分别为0.053μmol/mL和0.135μmol/mL，呈单层吸附排列。蛋白质解吸效率大于95%，这意味着制备的IMAM可重复用于蛋白质吸附。
