Rufo Joseph, Zhang Peiran, Wang Zeyu, Gu Yuyang, Yang Kaichun, Rich Joseph, Chen Chuyi, Zhong Ruoyu, Jin Ke, He Ye, Xia Jianping, Li Ke, Wu Jiarong, Ouyang Yingshi, Sadovsky Yoel, Lee Luke P, Huang Tony Jun
Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC USA.
Department of Biomedical Engineering, Duke University, Durham, NC USA.
Microsyst Nanoeng. 2024 Feb 4;10:23. doi: 10.1038/s41378-023-00648-3. eCollection 2024.
Extracellular vesicles (EVs) have been identified as promising biomarkers for the noninvasive diagnosis of various diseases. However, challenges in separating EVs from soluble proteins have resulted in variable EV recovery rates and low purities. Here, we report a high-yield ( > 90%) and rapid ( < 10 min) EV isolation method called FLocculation via Orbital Acoustic Trapping (FLOAT). The FLOAT approach utilizes an acoustofluidic droplet centrifuge to rotate and controllably heat liquid droplets. By adding a thermoresponsive polymer flocculant, nanoparticles as small as 20 nm can be rapidly and selectively concentrated at the center of the droplet. We demonstrate the ability of FLOAT to separate urinary EVs from the highly abundant Tamm-Horsfall protein, addressing a significant obstacle in the development of EV-based liquid biopsies. Due to its high-yield nature, FLOAT reduces biofluid starting volume requirements by a factor of 100 (from 20 mL to 200 µL), demonstrating its promising potential in point-of-care diagnostics.
细胞外囊泡（EVs）已被确认为各种疾病无创诊断的有前景的生物标志物。然而，将EVs与可溶性蛋白质分离面临挑战，导致EVs回收率各异且纯度较低。在此，我们报告一种名为“通过轨道声阱絮凝法分离细胞外囊泡（FLOAT）”的高产率（>90%）且快速（<10分钟）的EVs分离方法。FLOAT方法利用声流体液滴离心机旋转并可控加热液滴。通过添加热响应性聚合物絮凝剂，小至20纳米的纳米颗粒可快速且选择性地浓缩在液滴中心。我们展示了FLOAT从高度丰富的Tamm-Horsfall蛋白中分离尿液EVs的能力，解决了基于EVs的液体活检开发中的一个重大障碍。由于其高产率特性，FLOAT将生物流体起始体积需求降低了100倍（从20毫升降至200微升），证明了其在即时诊断中的广阔潜力。
