Nakano Asuka, Camacho-Alanis Fernanda, Ros Alexandra
Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
Analyst. 2015 Feb 7;140(3):860-8. doi: 10.1039/c4an01503g.
Insulator-based dielectrophoresis (iDEP) has been explored as a powerful analytical technique in recent years. Unlike with larger entities such as cells, bacteria or organelles, the mechanism of iDEP transport of proteins remains little explored. In this work, we extended the pool of proteins investigated with iDEP in nanostructured devices with β-galactosidase. Our work indicates that β-galactosidase shows concentration due to negative DEP which we compare to DEP response of immunoglobulin G (IgG) encapsulated in micelles also showing negative DEP. Experimental observations are further compared with numerical simulations to elucidate the influence of electrokinetic transport and the magnitude of DEP mobility. Numerical simulations suggest that the DEP mobility calculated using the classical model underestimates the actual contribution of DEP on the experimentally monitored concentration effect of proteins. Moreover, we observed a unique voltage dependent β-galactosidase concentration which we attribute to an additional factor influencing the protein concentration at the nanoconstrictions, namely ion concentration polarization. Our work aids in understanding factors influencing protein iDEP transport which is required for the future development of protein preconcentration or separation methods based on iDEP.
近年来,基于绝缘体的介电电泳(iDEP)已被探索作为一种强大的分析技术。与细胞、细菌或细胞器等较大的实体不同,蛋白质的iDEP传输机制仍鲜为人知。在这项工作中,我们在具有β-半乳糖苷酶的纳米结构装置中,扩展了用iDEP研究的蛋白质种类。我们的工作表明,β-半乳糖苷酶由于负介电电泳而出现浓度变化,我们将其与包裹在胶束中的免疫球蛋白G(IgG)的介电电泳响应进行比较,后者也显示出负介电电泳。实验观察结果进一步与数值模拟进行比较,以阐明电动传输的影响和介电电泳迁移率的大小。数值模拟表明,使用经典模型计算的介电电泳迁移率低估了介电电泳对实验监测的蛋白质浓度效应的实际贡献。此外,我们观察到一种独特的电压依赖性β-半乳糖苷酶浓度,我们将其归因于影响纳米收缩处蛋白质浓度的另一个因素,即离子浓度极化。我们的工作有助于理解影响蛋白质iDEP传输的因素,这是基于iDEP的蛋白质预浓缩或分离方法未来发展所必需的。