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梯度导电电极的蛋白质介电泳为经验理论提供了新的视角。

Protein Dielectrophoresis with Gradient Array of Conductive Electrodes Sheds New Light on Empirical Theory.

机构信息

Nanophotonics and Metrology Laboratory (NAM), Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne1015, Switzerland.

The Polytechnic School, Arizona State University, Mesa, Arizona85212, United States.

出版信息

Anal Chem. 2023 Feb 7;95(5):2958-2966. doi: 10.1021/acs.analchem.2c04708. Epub 2023 Jan 24.

Abstract

Dielectrophoresis (DEP) is a versatile tool for the precise microscale manipulation of a broad range of substances. To unleash the full potential of DEP for the manipulation of complex molecular-sized particulates such as proteins requires the development of appropriate theoretical models and their comprehensive experimental verification. Here, we construct an original DEP platform and test the Hölzel-Pethig empirical model for protein DEP. Three different proteins are studied: lysozyme, BSA, and lactoferrin. Their molecular Clausius-Mossotti function is obtained by detecting their trapping event via the measurement of the fluorescence intensity to identify the minimum electric field gradient required to overcome dispersive forces. We observe a significant discrepancy with published theoretical data and, after a very careful analysis to rule out experimental errors, conclude that more sophisticated theoretical models are required for the response of molecular entities in DEP fields. The developed experimental platform, which includes arrays of sawtooth metal electrode pairs with varying gaps and produces variations of the electric field gradient, provides a versatile tool that can broaden the utilization of DEP for molecular entities.

摘要

介电泳(DEP)是一种用于精确微尺度操控各种物质的多功能工具。为了充分发挥DEP 在操控复杂分子大小的颗粒(如蛋白质)方面的潜力,需要开发适当的理论模型并对其进行全面的实验验证。在这里,我们构建了一个原始的DEP 平台,并测试了 Hölzel-Pethig 蛋白质 DEP 的经验模型。研究了三种不同的蛋白质:溶菌酶、BSA 和乳铁蛋白。通过检测它们的荧光强度来识别克服分散力所需的最小电场梯度,从而获得它们的分子 Clausius-Mossotti 函数。我们观察到与已发表的理论数据有很大差异,经过非常仔细的分析排除了实验误差,我们得出结论,对于 DEP 场中分子实体的响应,需要更复杂的理论模型。所开发的实验平台包括具有不同间隙的锯齿状金属电极对阵列,可产生电场梯度的变化,提供了一种多功能工具,可拓宽DEP 在分子实体中的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3d33/9909730/d364e00223c4/ac2c04708_0002.jpg

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