磷酰胆碱色谱固定相的制备及其在分离生物大分子中的应用

以三氯氧磷、氯化胆碱和甲基丙烯酸-2-羟乙酯(HEMA)为原料合成了含磷酰胆碱的单体2-甲基丙烯酰氧乙基磷酰胆碱(MPC). 在硅胶表面嫁接聚合MPC, 得到磷酰胆碱两性离子交换色谱填料. 研究了该填料对标准蛋白的分离性能及pH对蛋白质保留的影响. 结果表明, 该填料对溶菌酶和牛血清白蛋白的动态吸附容量分别为13.8和18.7 mg/g, 其基质磷酰胆碱色谱固定相可同时基线分离两种酸性和两种碱性蛋白.

Preparation of Phosphocholine Chromatographic Stationary Phases and Their Application to Separation of Proteins

2-Methacryloyloxy ethyl phosphorylcholine(MPC) was prepared by the reaction of phosphorylcholine dichloride with 2-hydroxyethyl methacrylate(HEMA).The novel phosphocholine chromatographic stationary phase was synthesized by free radical graft polymerization of MPC from the surface of porous silica particles.The polymerization was initiated by thermal cleavage of tert-butylperoxy groups covalently attached to the particle surface.The stationary phase was evaluated in detail to determine its separability and to study the effect of pH on the retention of proteins.The dynamic protein loading capacity of the synthesized phosphocholine chromatographic stationary phase for Lys and BSA was 13.8 and 18.7 mg/g,respectively.The experimental results show that two acid proteins and two basic proteins were separated with 10 min linear gradient elution via the synthesized phosphocholine stationary phase.