Large-scale, cross-flow based isolation of highly pure and endocytosis-competent extracellular vesicles

Journal of Extracellular Vesicles

Carolina P. Caro-Vegas¹  Justin T. Landis¹  Dirk P. Dittmer¹  Lindsey M. Costantini¹  Ryan P. McNamara¹  Jack D. Griffith¹  Blossom A. Damania¹ 

[1] The University of North Carolina;

关键词: Extracellular vesicles;    industrial scale;    tangential flow;    cross-flow;    capto core;   

DOI  :  10.1080/20013078.2018.1541396

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown