以涂敷型硅胶为基质的弱阳离子交换色谱固定相的制备及其在生物大分子分离中的应用

在大孔硅胶表面键合乙烯基硅烷化试剂，然后与甲基丙烯酸环氧丙酯、甲基丙烯酸乙二醇双酯共聚，制得聚合物涂敷硅胶，其表面经水解再与丁二酸酐反应，得到一种新型的弱阳离子交换色谱填料（Weak Cation Exchange，WCX），考察了该填料对标准蛋白的分离性能，以及流速、pH值对蛋白质保留的影响．实验结果表明，在流速为3mL／min时，采用线性梯度洗脱，2min内可快速分离肌红蛋白、核糖核酸酶、细胞色素-C和溶菌酶4种标准碱性蛋白质，蛋白质的保留符合阳离子交换色谱保留规律．

Preparation of A Weak Cation Exchange Packing Based on Coated Macroporous Silica and Application in Protein Separation

A new macroporous silica-coated weak cation exchange packings was synthesized in following steps. First, vinyltriethoxysilane was bonded on silica with hydroxyl group. Second, vinyl-silica was copolymerized with glycidylmethacrylate and ethtlenedimethacrylate. Third, glycidylmethacrylate- ethtlenedimethacrylate coated silica was dispersed in succinic anhydride of pyridine solution. The effect of pH of mobile phase and flow rate on the protein retention and the packing characteristics in the separation of standard proteins were researched. The obtained results show that four standard proteins such as Myo, RNase-A,Cyt-C and Lys were completely separated by the linear gradient elution in 2 min at flow rate of 3 mL/min. The proteins retention accord with the retention rule of cation exchange chromatography. The packings is a better alternative for the biopolymer separation.