Extracellular vesicles (EVs) are heterogeneous membranous vesicles secreted by every cell type and offer significant potential in therapy and diagnostics. Differential ultracentrifugation is the gold standard for EV isolation, although other techniques including, polyethylene glycol (PEG) precipitation, immunoprecipitation, size exclusion chromatography, and immuno‐isolation approaches are common. Purified EVs can be characterized based on their physical characteristics, biochemical composition, or cell of origin. For size and concentration measurement, nanoparticle tracking analysis (NTA), dynamic light scattering (DLS), and electron microscopy are commonly employed methods. Biochemical analyses of EVs are typically performed using flow cytometry, immunoblotting, or proteomic investigation. Based on tissue of origin, EVs have specific markers that can be used to isolate and purify specific cell‐associated EVs using an affinity selection approach. Despite existence of several methods for isolation and characterization, major limitations associated with each method hinder the progress of the field. Evolving concepts in EV biology possess great promise for better isolation and characterization leading to a better insight of biological function and have immense clinical implications. In this review, we discuss recent advancements in EV isolation and characterization approaches.

中文翻译：

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细胞外囊泡的制备和表征

细胞外囊泡 (EV) 是由每种细胞类型分泌的异质膜囊泡，在治疗和诊断方面具有巨大潜力。差速超速离心是 EV 分离的金标准，但其他技术包括聚乙二醇 (PEG) 沉淀、免疫沉淀、尺寸排阻色谱和免疫分离方法也很常见。纯化的 EV 可以根据其物理特性、生化成分或来源细胞进行表征。对于尺寸和浓度测量，纳米粒子跟踪分析 (NTA)、动态光散射 (DLS) 和电子显微镜是常用的方法。EV 的生化分析通常使用流式细胞术、免疫印迹或蛋白质组学研究进行。根据起源组织，EV 具有特定的标记，可用于使用亲和选择方法分离和纯化特定的细胞相关 EV。尽管存在几种用于隔离和表征的方法，但与每种方法相关的主要限制阻碍了该领域的进展。EV 生物学中不断发展的概念对更好的分离和表征具有很大的希望，从而更好地了解生物功能，并具有巨大的临床意义。在这篇综述中，我们讨论了 EV 隔离和表征方法的最新进展。EV 生物学中不断发展的概念对更好的分离和表征具有很大的希望，从而更好地了解生物功能并具有巨大的临床意义。在这篇综述中，我们讨论了 EV 隔离和表征方法的最新进展。EV 生物学中不断发展的概念对更好的分离和表征具有很大的希望，从而更好地了解生物功能并具有巨大的临床意义。在这篇综述中，我们讨论了 EV 隔离和表征方法的最新进展。