Isolation of extracellular vesicles (EVs) from cell culture supernatant or plasma can be accomplished in a variety of ways. Common measures to quantify relative success are: concentration of the EVs, purity from non-EVs associated protein, size homogeneity and functionality of the final product. Here, we present an industrial-scale workflow for isolating highly pure and functional EVs using cross-flow based filtration coupled with high-molecular weight Capto Core size exclusion. Through this combination, EVs loss is kept to a minimum. It outperforms other isolation procedures based on a number of biochemical and biophysical assays. Moreover, EVs isolated through this method can be further concentrated down or directly immunopurified to obtain discreet populations of EVs. From our results, we propose that cross-flow/Capto Core isolation is a robust method of purifying highly concentrated, homogenous, and functionally active EVs from industrial-scale input volumes with few contaminants relative to other methods.

从细胞培养上清液或血浆中分离细胞外囊泡（EVs）可以通过多种方式完成。量化相对成功的常用方法是：电动汽车的浓度，非电动汽车相关蛋白的纯度，最终产品的尺寸均质性和功能性。在这里，我们介绍了一种工业规模的工作流程，该工作流程使用基于交叉流的过滤技术以及高分子量Capto Core尺寸排除技术来隔离高纯度和功能性的EV。通过这种结合，可以将电动汽车的损失降至最低。它优于基于许多生化和生物物理分析的其他分离程序。而且，通过这种方法分离出的电动汽车可以进一步浓缩或直接免疫纯化以获得离散的电动汽车群体。根据我们的结果，