利用辅助识别聚合物链制备牛血清白蛋白分子印迹聚合物

论文提出了一种新的蛋白质分子印迹方法, 即以聚乙烯醇接枝聚合物作为辅助识别聚合物链(ARPCs), 以丙烯酰胺为单体, 在丙烯酸酯树脂载体表面进行聚合, 制备牛血清白蛋白分子印迹树脂. 实验使用2.00 mol•L⁻¹氯化钾(KC1)溶液除去模板蛋白质, 使用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)检测吸附蛋白质, 并设计使用0.150, 0.500和2.00 mol•L⁻¹KC1溶液分别对吸附蛋白质进行梯度洗提. 电泳实验结果表明, 2.00 mol•L⁻¹KC1溶液洗脱的蛋白质是大孔树脂的吸附效应; 0.500 mol•L⁻¹KC1溶液洗脱掉的是印迹蛋白质. 引入ARPCs制得的分子印迹树脂, 用于混合蛋白质体系吸附时, 对模板蛋白质的吸附量为80~100μg•g⁻¹, 其特异性吸附能力较未引入ARPCs之前有明显提高.

Molecularly imprinted polymer with assistant recognition polymer chains for bovine serum albumin

A new protein molecularly imprinted polymer (MIP) was prepared with grafting polyvinyl alcohol as assistant recognition polymer chains (ARPCs). The ARPCs and acrylamide monomers were interpenetrated and then polymerized on the surface of macroporous acrylate adsorbent spheres. The template BSA was removed by treatment with 2.00 mol•L⁻¹ potassium chloride (KCl) solution and the adsorbed proteins were detected with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). 0.150 mol•L⁻¹, 0.500 mol•L⁻¹, and 2.00 mol•L⁻¹KCl solutions were used as eluent to wash the adsorbed proteins. The SDS-PAGE results show that proteins washed out with 2.00 mol•L⁻¹KCl solution were from nonspecific adsorption of macroporous acrylate adsorbent spheres, and proteins washed out with 0.500 mol•L⁻¹KC1 solution were specific proteins imprinted by MIP resins. MIP resins with ARPCs had better recognition to the target proteins than that without ARPCs. The adsorption capacity of MIP resins immobilized ARPCs to the template BSA was about 80-100μg•g⁻¹ when it was used for the adsorption of proteins mixture, and the specific adsorption of the target protein was obviously increased.