Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023:2668:45-55.
doi: 10.1007/978-1-0716-3203-1_5.

Extracellular Vesicle Isolation by a Tangential-Flow Filtration-Based Large-Scale Purification Method

Runjie Yuan  1 , Yijun Zhou  1 , Gabriel F Arias  2 , Dirk P Dittmer  3
Affiliations

Extracellular Vesicle Isolation by a Tangential-Flow Filtration-Based Large-Scale Purification Method

Runjie Yuan et al. Methods Mol Biol. 2023.

Abstract

Extracellular vesicle (EV) isolation from conditioned cell culture medium has been a challenging topic. It is particularly difficult to obtain pure and intact EVs at a large scale. The commonly used methods such as differential centrifugation, ultracentrifugation, size exclusion chromatography, polyethylene glycol (PEG) precipitation, filtration, and affinity-based purification each have their advantages and limitations. Here, we present a tangential-flow filtration (TFF) based, multi-step purification protocol that combines filtration, PEG precipitation, and Capto Core 700 multimodal chromatography (MMC) to isolate EVs at high purity from large volumes of cell culture conditioned medium. Inserting the TFF step before PEG precipitation removes proteins, which may aggregate in subsequent steps and co-purify with EVs.

Keywords: Capto Core 700 multimodal chromatography (MMC); Extracellular vesicles (EVs); Purification; Tangential-flow filtration (TFF).

PubMed Disclaimer

References

    1. Kalluri R, LeBleu VS (2020) The biology, function, and biomedical applications of exosomes. Science 367(6478):eaau6977 - DOI - PubMed - PMC
    1. Théry C, Witwer KW, Aikawa E et al (2018) Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 7(1):1535750 - DOI - PubMed - PMC
    1. Théry C, Amigorena S, Raposo G et al (2006. Chapter 3) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol 3:22
    1. Lobb RJ, Becker M, Wen SW et al (2015) Optimized exosome isolation protocol for cell culture supernatant and human plasma. J Extracell Vesicles 4:27031 - DOI - PubMed
    1. McNamara RP, Caro-Vegas CP, Costantini LM et al (2018) Large-scale, cross-flow based isolation of highly pure and endocytosis-competent extracellular vesicles. J Extracell Vesicles 7(1):1541396 - DOI - PubMed - PMC

Publication types

Substances

LinkOut - more resources