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Rapid Detection and Trapping of Extracellular Vesicles by Electrokinetic Concentration for Liquid Biopsy on Chip

机译:芯片液体活检电动浓度快速检测和诱捕细胞内囊泡

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摘要

Exosomes have gained immense importance since their proteomic and genetic contents could potentially be used for disease diagnostics, monitoring of cancer progression, metastasis, and drug efficacy. However, establishing the clinical utility of exosomes has been restricted due to small sizes and high sample loss from extensive sample preparation. Sample loss is particularly critical for body fluids limited in volume and difficult to access, e.g., cerebrospinal fluid. We present a microfluidic technique that locally enhances the concentration of extracellular vesicles extracted from MDA-MB-231 human breast cancer cell lines by using an ion concentration polarization (ICP)-based electrokinetic concentrator. Our design incorporates a trapping mechanism near the conductive polymer membrane; therefore, we can preconcentrate and capture extracellular vesicles simultaneously. Compared with standard fluorescence detection, our method increased the limit of detection (LOD) of extracellular vesicles by two orders of magnitude in 30 min. Our concentrator increased the extracellular vesicle concentration for 5.0 × 107 particles/1 mL (LOD), 5.0 × 108 particles/1 mL, and 5.0 × 109 particles/1 mL by ~100-fold each within 30 min using 45 V. This study demonstrates an alternative platform to simultaneously preconcentrate and capture extracellular vesicles that can be incorporated as part of a liquid biopsy-on-a-chip system for the detection of exosomal biomarkers and analysis of their contents for early cancer diagnosis.
机译:由于它们的蛋白质组学和遗传含量可能用于疾病诊断,监测癌症进展,转移和药物疗效,因此外泌体具有巨大的重要性。然而,由于广泛的样品制备,因此建立了外索体的临床用途受到限制性和高样品损失。对于体积限制并且难以进入,例如脑脊液,样品损失特别关键。我们介绍了一种微流体技术,通过使用离子浓度极化(ICP)的电动浓缩器,局部增强了从MDA-MB-231人乳腺癌细胞系中提取的细胞外囊的浓度。我们的设计采用了导电聚合物膜附近的捕获机制;因此,我们可以同时预先培育并捕获细胞外囊泡。与标准荧光检测相比,我们的方法在30分钟内将细胞外囊泡的检测(LOD)的序列增加了两个数量级。在使用45 V.本研究中,我们的浓缩器增加了5.0×107颗粒/ 1mL(LOD),5.0×108颗粒/ 1ml,5.0×108颗粒/ 1ml,5.0×109颗粒/ 1ml的〜100倍。该研究证明一种替代平台,以同时预肠化和捕获可以掺入的细胞外囊泡,该细胞外囊泡可以作为液检结果的含芯片系统的一部分掺入,用于检测外泌体生物标志物和其含量的早期癌症诊断的分析。

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