Extracellular vesicles (EVs) are heterogenous membrane-bound vesicles released from various origins. EVs play a crucial role in cellular communication and mediate several physiological and pathological processes, highlighting their potential therapeutic and diagnostic applications. Due to the rapid increase in interests and needs to elucidate EV properties and functions, numerous isolation and separation approaches for EVs have been developed to overcome limitations of conventional techniques, such as ultracentrifugation. This review focuses on recently emerging and modern EV isolation and separation techniques, including size-, charge-, and affinity-based techniques while excluding ultracentrifugation and precipitation-based techniques due to their multiple limitations. The advantages and drawbacks of each technique are discussed together with insights into their applications. Emerging approaches all share similar features in terms of being time-effective, easy-to-operate, and capable of providing EVs with suitable and desirable purity and integrity for applications of interest. Combination and hyphenation of techniques have been used for EV isolation and separation to yield EVs with the best quality. The most recent development using an automated on-line system including selective affinity-based trapping unit and asymmetrical flow field-flow fractionation allows reliable isolation and fractionation of EV subpopulations from human plasma.

细胞外囊泡（EVs）是从各种来源释放的异质膜结合囊泡。电动汽车在细胞通讯中起着至关重要的作用，并介导了几种生理和病理过程，突出了其潜在的治疗和诊断应用。由于兴趣的快速增长和阐明电动汽车特性和功能的需求，因此，为克​​服传统技术（如超速离心）的局限性，开发了许多电动汽车的隔离和分离方法。这篇综述着重于新兴的和现代的电动汽车隔离技术，包括基于大小，电荷和亲和力的技术，同时由于其多重局限性而排除了超速离心和基于沉淀的技术。讨论了每种技术的优缺点以及对它们的应用的见解。新兴方法在时间效率，易于操作以及能够为感兴趣的应用提供具有合适和所需纯度和完整性的电动汽车方面均具有相似的功能。EV的隔离和分离已采用了技术的组合和连字技术，以产生具有最佳质量的EV。使用包括基于选择性亲和力的捕获单元和不对称流场-流分离的自动化在线系统的最新开发，可以从人血浆中可靠地分离和分离EV亚群。并能够为感兴趣的应用提供具有合适且理想的纯度和完整性的电动汽车。EV的隔离和分离已采用了技术的组合和连字符，以产生具有最佳质量的EV。使用包括基于选择性亲和力的捕获单元和不对称流场-流分离的自动化在线系统的最新开发，可以从人血浆中可靠地分离和分离EV亚群。并能够为感兴趣的应用提供具有合适且理想的纯度和完整性的电动汽车。EV的隔离和分离已采用了技术的组合和连字技术，以产生具有最佳质量的EV。使用包括基于选择性亲和力的捕获单元和不对称流场-流分离的自动化在线系统的最新开发，可以从人血浆中可靠地分离和分离EV亚群。