Procedures for enrichment, isolation and purification of exosomes from complex biological samples are difficult, tedious, non-standardized, and require bulky instrumentation such as ultracentrifugation (UC). In this article, a simple method for isolating exosomes. Size-Selective Method (SSM) was established based on commercially available materials, and the UC and ExoQuick-TC kits (EQkit) methods were compared in terms of morphology, particle size, quantity, Western Blot (WB), and extraction time. Results showed that all three different exosome separation methods could obtain circular membranous vesicles, with a diameter of 30-110 nm. There were more non-exosome components in the samples extracted by SSM, such as large microvesicles, with a lower purity. UC obtained a large number of exosomes with a higher purity, but it required an ultracentrifuge, costed much time and had low yield. Both EQkit and SSM were easy to operate, but EQkit tended to aggregate exosomes and consume much time. WB results showed that exosomes extracted by all the three methods expressed CD63 protein. The SSM had the highest CD63 protein content and at the same protein concentration. The above evidences showed that SSM was fast and had high recovery, low cost and high protein concentration, but had more non-exosome protein components, which can be a choice for exosome separation.

中文翻译：

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建立并评估外泌体富集和纯化的简单大小选择方法。

从复杂的生物样品中富集，分离和纯化外泌体的过程困难，繁琐，不标准化，并且需要笨重的仪器，例如超速离心（UC）。在本文中，一种分离外泌体的简单方法。基于市售材料建立了尺寸选择方法（SSM），并比较了UC和ExoQuick-TC试剂盒（EQkit）的形态，粒径，数量，蛋白质印迹（WB）和提取时间。结果表明，三种不同的外泌体分离方法均可获得直径为30-110 nm的圆形膜状囊泡。用SSM提取的样品中有更多的非外泌体成分，例如较大的微泡，且纯度较低。UC获得了大量纯度更高的外泌体，但它需要超速离心机，花费大量时间且产量低。EQkit和SSM都易于操作，但EQkit倾向于聚集外泌体并消耗大量时间。WB结果表明，三种方法提取的外泌体均表达CD63蛋白。SSM具有最高的CD63蛋白质含量，且蛋白质浓度相同。以上证据表明，SSM具有快速，回收率高，成本低，蛋白浓度高等优点，但非外泌体蛋白成分较多，可以作为外泌体分离的选择。SSM具有最高的CD63蛋白质含量，且蛋白质浓度相同。以上证据表明，SSM具有快速，回收率高，成本低，蛋白浓度高等优点，但非外泌体蛋白成分较多，可以作为外泌体分离的选择。SSM具有最高的CD63蛋白质含量，且蛋白质浓度相同。以上证据表明，SSM具有快速，回收率高，成本低，蛋白浓度高等优点，但非外泌体蛋白成分较多，可以作为外泌体分离的选择。