Asghar-Lab-Micro and Nanotechnology in Medicine Lab, Florida Atlantic University, Boca Raton, FL 33431, USA.
Department of Biological Sciences, Florida Atlantic University, Boca Raton, FL 33431, USA.
Sensors (Basel). 2023 Oct 7;23(19):8292. doi: 10.3390/s23198292.
Exosomes have gained recognition in cancer diagnostics and therapeutics. However, most exosome isolation methods are time-consuming, costly, and require bulky equipment, rendering them unsuitable for point-of-care (POC) settings. Microfluidics can be the key to solving these challenges. Here, we present a double filtration microfluidic device that can rapidly isolate exosomes via size-exclusion principles in POC settings. The device can efficiently isolate exosomes from 50-100 µL of plasma within 50 min. The device was compared against an already established exosome isolation method, polyethylene glycol (PEG)-based precipitation. The findings showed that both methods yield comparable exosome sizes and purity; however, exosomes isolated from the device exhibited an earlier miRNA detection compared to exosomes obtained from the PEG-based isolation. A comparative analysis of exosomes collected from membrane filters with 15 nm and 30 nm pore sizes showed a similarity in exosome size and miRNA detection, with significantly increased sample purity. Finally, TEM images were taken to analyze how the developed devices and PEG-based isolation alter exosome morphology and to analyze exosome sizes. This developed microfluidic device is cost-efficient and time-efficient. Thus, it is ideal for use in low-resourced and POC settings to aid in cancer and disease diagnostics and therapeutics.
外泌体在癌症诊断和治疗中得到了认可。然而,大多数外泌体分离方法耗时、昂贵,且需要大型设备,因此不适合即时检测(POC)环境。微流控技术可能是解决这些挑战的关键。在这里,我们提出了一种双重过滤微流控装置,可通过尺寸排阻原理在 POC 环境中快速分离外泌体。该装置可在 50 分钟内从 50-100μL 的血浆中有效分离外泌体。该装置与已建立的外泌体分离方法——聚乙二醇(PEG)沉淀进行了比较。结果表明,两种方法得到的外泌体大小和纯度相当;然而,与从 PEG 分离得到的外泌体相比,从该装置分离得到的外泌体更早检测到 miRNA。对 15nm 和 30nm 孔径的膜过滤器收集的外泌体进行比较分析表明,外泌体大小和 miRNA 检测具有相似性,且样品纯度显著提高。最后,通过 TEM 图像分析了开发的设备和基于 PEG 的分离如何改变外泌体的形态并分析外泌体的大小。这种开发的微流控装置具有成本效益和时间效益。因此,它非常适合在资源有限和 POC 环境中使用,以辅助癌症和疾病的诊断和治疗。