Direct isolation of small extracellular vesicles from human blood using viscoelastic microfluidics.

Direct isolation of small extracellular vesicles from human blood using viscoelastic microfluidics.
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DOI:
10.1126/sciadv.adi5296
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发表时间:
2023-10-06
期刊:
影响因子:
13.6
通讯作者:
deMello, Andrew J.
deMello, Andrew J.
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Meng, Yingchao;Zhang, Yanan;Buhler, Marcel;Wang, Shuchen;Asghari, Mohammad;Sturchler, Alessandra;Mateescu, Bogdan;Weiss, Tobias;Stavrakis, Stavros;deMello, Andrew J.

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含有脂质、核酸和蛋白质的小细胞外囊泡(sEV; <200 nm)被认为是多种疾病的有希望的生物标志物。从血液中分离sEV的常规方法与常规临床工作流程不相容,显著阻碍了血液来源的sEV在临床环境中的利用。在这里,我们提出了一个简单的,基于粘弹性的微流体平台,用于从人血液中无标记分离sEV。通过从全血中分离荧光sEV来评估该装置的分离性能,证明纯度和回收率分别超过97%和87%。值得注意的是,与金标准超离心相比,我们的基于粘弹性的微流体方法还提供了sEV产率的显着增加,通过两种方法纯化的血液来源的sEV的蛋白质组学谱显示出相似的蛋白质组成。为了证明该方法的临床实用性,我们从20名癌症患者和20名健康供体的血液样本中分离出sEV,证明在来自癌症患者的血液中可以观察到升高的sEV浓度。粘弹性微流体能够以高分离效率和回收率从人血液中无标记地分离sEV。
Small extracellular vesicles (sEVs; <200 nm) that contain lipids, nucleic acids, and proteins are considered promising biomarkers for a wide variety of diseases. Conventional methods for sEV isolation from blood are incompatible with routine clinical workflows, significantly hampering the utilization of blood-derived sEVs in clinical settings. Here, we present a simple, viscoelastic-based microfluidic platform for label-free isolation of sEVs from human blood. The separation performance of the device is assessed by isolating fluorescent sEVs from whole blood, demonstrating purities and recovery rates of over 97 and 87%, respectively. Significantly, our viscoelastic-based microfluidic method also provides for a remarkable increase in sEV yield compared to gold-standard ultracentrifugation, with proteomic profiles of blood-derived sEVs purified by both methods showing similar protein compositions. To demonstrate the clinical utility of the approach, we isolate sEVs from blood samples of 20 patients with cancer and 20 healthy donors, demonstrating that elevated sEV concentrations can be observed in blood derived from patients with cancer. Viscoelastic microfluidics enables label-free isolation of sEVs from human blood with high separation efficiency and recovery.
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