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Cited by 308

The DOI matches its Crossref record; 74 of the 74 references it cites were found in a registry or an index.

Comparative analysis of tangential flow filtration and ultracentrifugation, both combined with subsequent size exclusion chromatography, for the isolation of small extracellular vesicles

Open access
Visan, Kekoolani S., Lobb, Richard J., Ham, Sunyoung, Lima, Luize G., Palma, Carlos, Edna, Chai Pei Zhi, Wu, Li‐Ying, Gowda, Harsha, Datta, Keshava K., Hartel, Gunter, Salomon, Carlos, Möller, Andreas·2022·Journal of Extracellular Vesicles

Abstract

Small extracellular vesicles (sEVs) provide major promise for advances in cancer diagnostics, prognostics, and therapeutics, ascribed to their distinctive cargo reflective of pathophysiological status, active involvement in intercellular communication, as well as their ubiquity and stability in bodily fluids. As a result, the field of sEV research has expanded exponentially. Nevertheless, there is a lack of standardisation in methods for sEV isolation from cells grown in serum‐containing media. The majority of researchers use serum‐containing media for sEV harvest and employ ultracentrifugation as the primary isolation method. Ultracentrifugation is inefficient as it is devoid of the capacity to isolate high sEV yields without contamination of non‐sEV materials or disruption of sEV integrity. We comprehensively evaluated a protocol using tangential flow filtration and size exclusion chromatography to isolate sEVs from a variety of human and murine cancer cell lines, including HeLa, MDA‐MB‐231, EO771 and B16F10. We directly compared the performance of traditional ultracentrifugation and tangential flow filtration methods, that had undergone further purification by size exclusion chromatography, in their capacity to separate sEVs, and rigorously characterised sEV properties using multiple quantification devices, protein analyses and both image and nano‐flow cytometry. Ultracentrifugation and tangential flow filtration both enrich consistent sEV populations, with similar size distributions of particles ranging up to 200 nm. However, tangential flow filtration exceeds ultracentrifugation in isolating significantly higher yields of sEVs, making it more suitable for large‐scale research applications. Our results demonstrate that tangential flow filtration is a reliable and robust sEV isolation approach that surpasses ultracentrifugation in yield, reproducibility, time, costs and scalability. These advantages allow for implementation in comprehensive research applications and downstream investigations.

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Crossref · 10.1002/jev2.12266

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Metadata

Authors
Visan, K. S., Lobb, R. J., Ham, S., Lima, L. G., Palma, C., Edna, C. P. Z., Wu, L., Gowda, H., Datta, K. K., Hartel, G., Salomon, C., & Möller, A.
Venue
Journal of Extracellular Vesicles
Publisher
Wiley
Published
2022-09
Volume
11
Issue
9
Pages
e12266-e12266
ISSN
2001-3078 (ISSN-L)
OpenAlex
W4296805213
Cited by
308
References
74
FWCI
14.29
Type
journal article
Language
en
Open access
gold

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Visan, K. S., Lobb, R. J., Ham, S., Lima, L. G., Palma, C., Edna, C. P. Z., Wu, L., Gowda, H., Datta, K. K., Hartel, G., Salomon, C., & Möller, A. (2022). Comparative analysis of tangential flow filtration and ultracentrifugation, both combined with subsequent size exclusion chromatography, for the isolation of small extracellular vesicles. Journal of Extracellular Vesicles. https://doi.org/10.1002/jev2.12266

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Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.