【摘要】 细胞外囊泡(EVs)是细胞向外部环境释放的膜结构颗粒，广泛存在于血液、尿液和唾液等体液中，包含蛋白质及核酸等多种生物分子，可作为多种疾病的生物标记物，其提取与分析对疾病的快速诊断与治疗至关重要。传统的EVs分选技术，如差速超速离心，存在样本需求量大且仪器设备昂贵等局限，而微流控分选技术因其微型化、高回收率和集成化等优势，更适合于医疗机构的实际应用。文中将微流控分选方法分类为无标记被动分选、无标记主动分选、固定基底免疫亲和分选以及游离微珠免疫亲和分选，并综述了各类方法在EVs分离领域的研究进展，概括和比较了各类方法在纯度、回收率、通量以及芯片制备难度等方面的差异和特点，展望了EVs微流控分选技术的发展方向，包括改善分选指标、降低分选成本和减小分离截止尺寸等改进目标。在无标记分选中，声泳具有纯度高、通量高以及可分选纳米EVs的优势，且能单片集成EVs浓度的声学传感模块，更具发展前景；而免疫亲和分选，因使用游离微珠技术，凭借其捕获效率高且芯片可复用的优势，更适用于连续和批量化的分选应用。更多还原

【Abstract】 Extracellular vesicles(EVs) are one type of membrane-enclosed particles released by cells into the extracellular environment. EVs are ubiquitously present in various body fluids including blood and urine. They serve as biomarkers for numerous diseases, owing to their content of diverse biomolecules like proteins and nucleic acids. The extraction and analysis of EVs are crucial for the rapid diagnosis and treatment of diseases. Traditional techniques employed for sorting EVs, such as differential ultracentrifugation, need a substantial amount of sample material and involve costly instrumentation. Conversely, microfluidic sorting techniques offer several benefits, including miniaturization, high recovery rates and integration, thus making them more suitable for the application in medical institutions and clinics. This review categorized microfluidic sorting methods into label-free passive, label-free active, fixed-substrate immunoaffinity and free-bead immunoaffinity sorting, introduced the research progress of these methods in the field of EVs isolation. The characteristics including purity, recovery, flux and difficulty in chip fabrication of each method are summarized and compared in the review. In addition, the review envisaged the future development directions for microfluidic sorting of EVs, which included sorting indexes improvement, sorting costs reduction and separation cut-off size decrement. For label-free sorting, acoustophoresis is more promising due to the advantages in achieving high purity and throughput, as well as the capability to sort nano-sized EVs. Acoustophoresis also enables monolithic integration of acoustic sensing modules for EV concentration. For immunoaffinity sorting, the technology using free beads has advantages of high capture efficiency and chip reusability, which is suitable for continuous and batch sorting scenarios.更多还原