CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, PR China.
CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, PR China.
Talanta. 2017 Aug 1;170:275-285. doi: 10.1016/j.talanta.2017.04.012. Epub 2017 Apr 7.
In this work, an antifouling capillary modified with star-shaped poly(2-methyl-2-oxazoline)-based copolymer was used to study the interaction between acetaminophen (APAP) and bovine serum albumin (BSA) by frontal analysis capillary electrophoresis (FACE). The star-shaped copolymer, poly(ethylene imine)-graft-poly(2-methyl-2-oxazoline) (PEI-g-PMOXA), was immobilized onto the fused-silica capillary inner wall via dopamine-assisted co-deposition strategy, yielding a PEI-g-PMOXA/polydopamine (PDA)-coated antifouling capillary, i.e., an antifouling capillary coated with the PEI-g-PMOXA/PDA co-deposited film. Electroosmotic flow (EOF) mobility of the PEI-g-PMOXA/PDA-coated capillary was almost zero in a wide pH range (3.0-10.0), while the EOF mobility of bare capillary was much larger and increased significantly with pH increasing. When the PEI-g-PMOXA/PDA-coated capillary was exploited to separate a protein mixture including cytochrome c, lysozyme, ribonuclease A and α-chymotrypsinogen A, the theoretical plate numbers were of five orders of magnitude which were about ten-fold higher over those obtained with bare capillary; in addition, the RSD values of migration time were mostly less than 0.7% (30 consecutive runs) which were much smaller than those of bare capillary (c.a. 5.7%). The protein-resistant PEI-g-PMOXA/PDA-coated capillary was then used to investigate the interaction between APAP and BSA by FACE, the binding constant and number of binding sites at 25°C and pH 7.4 (Tris/HCl buffer of 25mM) were 1.39×10M and 1.08, respectively, which were comparable to the results determined by fluorescence spectroscopic measurement (3.18×10M and 1.19, respectively).
在这项工作中,使用了一种经过星形聚(2-甲基-2-恶唑啉)基共聚物修饰的抗污毛细管,通过前沿分析毛细管电泳(FACE)研究了对乙酰氨基酚(APAP)与牛血清白蛋白(BSA)之间的相互作用。星形共聚物,聚乙烯亚胺-接枝-聚(2-甲基-2-恶唑啉)(PEI-g-PMOXA),通过多巴胺辅助共沉积策略固定在熔融石英毛细管内壁上,得到一种 PEI-g-PMOXA/聚多巴胺(PDA)涂层的抗污毛细管,即一种 PEI-g-PMOXA/PDA 共沉积膜涂覆的抗污毛细管。在很宽的 pH 范围(3.0-10.0)内,PEI-g-PMOXA/PDA 涂层毛细管的电渗流(EOF)迁移率几乎为零,而裸毛细管的 EOF 迁移率则大得多,并随 pH 值的增加而显著增加。当 PEI-g-PMOXA/PDA 涂层毛细管用于分离包括细胞色素 c、溶菌酶、核糖核酸酶 A 和 α-糜蛋白酶原 A 的蛋白质混合物时,理论板数达到了五个数量级,比裸毛细管高约十倍;此外,迁移时间的 RSD 值大多小于 0.7%(30 次连续运行),比裸毛细管小得多(约 5.7%)。然后,使用具有蛋白质抗性的 PEI-g-PMOXA/PDA 涂层毛细管通过 FACE 研究 APAP 与 BSA 之间的相互作用,在 25°C 和 pH 7.4(25mM Tris/HCl 缓冲液)下的结合常数和结合位点数分别为 1.39×10^M 和 1.08,这与荧光光谱测量确定的结果相当(分别为 3.18×10^M 和 1.19)。