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外泌体是由活细胞主动分泌的一种纳米级囊泡，内含蛋白质、核酸等生物活性物质，参与细胞间通讯，在肿瘤微环境调节、免疫识别、炎症反应等众多生理病理过程中发挥重要作用。作为一种运输载体，外泌体广泛存在于生物流体中，但由于其粒径小、内含物丰度低，能否获得高质量的外泌体成为后续研究成败的关键。虽然外泌体分离、纯化和鉴定方法日趋发展，但仍未达到成熟水平。本文就外泌体分离和纯化方法的研究进行综述，并比较不同方法的优缺点、适用范围及应用前景，以期为外泌体研究提供参考。“,”Exosomes are nano-scale vesicles and secreted by living cells actively, which contain proteins, nucleic acids and other bioactive substances, participate in intercellular communication, and play an important role in many physiological and pathological processes, such as tumor microenvironment regulation, immune recognition, inflammatory reaction and so on. As a transport carrier, exosomes exist abundantly in biological fluids. However, due to their small particle size and low abundance of inclusions, the acquirement of high-quality exosomes is critical for further experiments. Although the separation, purification and identification of exosomes have been developed, there are still a lot of work to be done. In this paper, the methods of exosome separation and purification were summarized with various advantages in application and prospect, providing some thoughts for the investigation of exosomes.