Chen Wenwen, Xie Yingying, Chang Yuang, Xu Yuhai, Zhao Mengqian, Deng Pengwei, Qin Jianhua, Li Hongjing
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
University of Chinese Academy of Sciences, Beijing 100864, China.
Micromachines (Basel). 2021 Sep 29;12(10):1182. doi: 10.3390/mi12101182.
Exosomes are membrane-bound nanovesicles secreted by most types of cells, which contain a series of biologically important molecules, such as miRNAs, proteins, and lipids, etc. Emerging evidence show that exosomes can affect the physiological status of cells and are involved in various pathological processes. However, due to their small size and density close to body fluids, it is challenging to separate exosomes from a small volume of biological samples in a simple manner. Herein, we propose a new strategy for isolating circulating exosomes from biological samples in a portable device. This method synergistically integrates chitosan electrostatic-adsorption, scaffold substrates, and shuttle flow to enable the highly effective capture of circulating exosomes with a recovery rate of over 80% within 20 min, which is much better than the performance of traditional ultracentrifugation (5-25%, 3 h). Besides, the isolated exosomes from samples could be lysed in situ and further subjected to RNA concentration detection and protein analysis. In particular, all the necessary procedures for exosome separation could be integrated into a single device without the need for bulky equipment. This established device is portable and easy to operate, which provides a promising platform for the study of exosome biology and clinical diagnosis.
外泌体是大多数类型细胞分泌的膜结合纳米囊泡，其包含一系列生物学上重要的分子，如微小RNA、蛋白质和脂质等。新出现的证据表明，外泌体可影响细胞的生理状态，并参与各种病理过程。然而，由于其体积小且密度接近体液，以简单的方式从少量生物样品中分离外泌体具有挑战性。在此，我们提出了一种在便携式设备中从生物样品中分离循环外泌体的新策略。该方法协同整合了壳聚糖静电吸附、支架底物和穿梭流，能够在20分钟内高效捕获循环外泌体，回收率超过80%，这比传统超速离心法（5-25%，3小时）的性能要好得多。此外，从样品中分离出的外泌体可原位裂解，并进一步进行RNA浓度检测和蛋白质分析。特别地，外泌体分离的所有必要程序都可整合到单个设备中，无需大型设备。所建立的设备便于携带且易于操作，为外泌体生物学研究和临床诊断提供了一个有前景的平台。
