Extracellular vesicles (EVs) play an important role in intercellular communication. Recently, there has been increasing interest in EVs as potential diagnostic biomarkers and therapeutic vehicles. However, the molecular properties and cargo information of EV subpopulations have not yet been fully investigated due to lack of reliable and reproducible EV separation technology. Current approaches have faced difficulties with efficiently isolating EVs from biofluids, especially subpopulations of small EVs. Here, we report an EV isolation method based on a size-selective microfluidic platform (ExoSMP) via nanomembrane filtration and electrophoretic force. This unique platform offers an enhanced approach to sorting a heterogeneous population of EVs based on size, with the additional advantages of being label-free and low-cost, and featuring a short processing time (<1 h), and convenient integration with downstream analysis. In this research, we used ExoSMP to demonstrate the isolation of cancer-derived small EVs (30–120 nm) with high recovery (94.2%) and reproducibility at an optimum sample flow rate. Furthermore, we investigated isolation of EV subpopulations by altering nanomembrane combinations with different pore size combinations (50 and 100 nm, 30 and 100 nm, 30 and 200 nm, and 30 and 50 nm). This ExoSMP technique can serve as a standardized EV isolation/separation tool, facilitating the clinical prospects of EVs and opening up a new avenue for future point-of-care applications in liquid biopsies.

中文翻译：

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通过尺寸选择性微流体平台分离癌症来源的细胞外囊泡亚群。

细胞外囊泡（EV）在细胞间通讯中发挥着重要作用。最近，人们对电动汽车作为潜在的诊断生物标志物和治疗工具越来越感兴趣。然而，由于缺乏可靠且可重复的 EV 分离技术，EV 亚群的分子特性和货物信息尚未得到充分研究。目前的方法在有效地将电动汽车与生物流体分离时面临着困难，特别是小型电动汽车的亚群。在这里，我们报告了一种基于尺寸选择性微流体平台（ExoSMP）通过纳米膜过滤和电泳力分离EV的方法。这个独特的平台提供了一种增强的方法来根据大小对不同种类的电动汽车进行分类，具有无标签和低成本的额外优势，并且处理时间短（<1小时），并且可以方便地与下游分析集成。在这项研究中，我们使用 ExoSMP 展示了在最佳样品流速下分离癌症衍生的小型 EV（30-120 nm）的高回收率（94.2%）和重现性。此外，我们通过改变具有不同孔径组合（50 和 100 nm、30 和 100 nm、30 和 200 nm、30 和 50 nm）的纳米膜组合来研究 EV 亚群的分离。这种 ExoSMP 技术可以作为标准化的 EV 隔离/分离工具，促进 EV 的临床前景，并为未来液体活检的护理点应用开辟新途径。