The DOI matches its Crossref record; 6 of the 6 references it cites were found in a registry or an index.
Heparin affinity purification of extracellular vesicles
Open accessAbstract
Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely used isolation method is ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs. Previously we have shown that heparin blocks EV uptake in cells, supporting a direct EV-heparin interaction. Here we show that EVs can be purified from cell culture media and human plasma using ultrafiltration (UF) followed by heparin-affinity beads. UF/heparin-purified EVs from cell culture displayed the EV marker Alix, contained a diverse RNA profile, had lower levels of protein contamination and were functional at binding to and uptake into cells. RNA yield was similar for EVs isolated by UC. We were able to detect mRNAs in plasma samples with comparable levels to UC samples. In conclusion, we have discovered a simple, scalable and effective method to purify EVs taking advantage of their heparin affinity.
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Cites
Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids
Théry, C., Amigorena, S., Raposo, G., & Clayton, A. (2006) · Current Protocols in Cell Biology · doi:10.1002/0471143030.cb0322s30 · Cited by 5,766
Christianson, H. C., Svensson, K. J., van Kuppevelt, T. H., Li, J.-P., & Belting, M. (2013) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1304266110 · Cited by 903
Webber, J., & Clayton, A. (2013) · Journal of Extracellular Vesicles · doi:10.3402/jev.v2i0.19861 · Cited by 817
Microfluidic isolation and transcriptome analysis of serum microvesicles
Chen, C., Skog, J., Hsu, C.-H., Lessard, R. T., Balaj, L., Wurdinger, T., Carter, B. S., Breakefield, X. O., Toner, M., & Irimia, D. (2010) · Lab Chip · doi:10.1039/b916199f · Cited by 572
Self-citationMicrovesicle-associated AAV Vector as a Novel Gene Delivery System
Maguire, C. A., Balaj, L., Sivaraman, S., Crommentuijn, M. H., Ericsson, M., Mincheva-Nilsson, L., Baranov, V., Gianni, D., Tannous, B. A., Sena-Esteves, M., et al. (2012) · Molecular Therapy · doi:10.1038/mt.2011.303 · Cited by 283
Self-citationNoerholm, M., Balaj, L., Limperg, T., Salehi, A., Zhu, L. D., Hochberg, F. H., Breakefield, X. O., Carter, B. S., & Skog, J. (2012) · BMC Cancer · doi:10.1186/1471-2407-12-22 · Cited by 190
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Cited by
Théry, C., Witwer, K. W., Aikawa, E., Alcaraz, M. J., Anderson, J. D., Andriantsitohaina, R., Antoniou, A., Arab, T., Archer, F., Atkin‐Smith, G. K., et al. (2018) · Journal of Extracellular Vesicles · doi:10.1080/20013078.2018.1535750 · Cited by 11,099
Self-citationWelsh, J. A., Goberdhan, D. C. I., O'Driscoll, L., Buzas, E. I., Blenkiron, C., Bussolati, B., Cai, H., Di Vizio, D., Driedonks, T. A. P., Erdbrügger, U., et al. (2024) · Journal of Extracellular Vesicles · doi:10.1002/jev2.12404 · Cited by 4,663
Extracellular vesicles in cancer — implications for future improvements in cancer care
Xu, R., Rai, A., Chen, M., Suwakulsiri, W., Greening, D. W., & Simpson, R. J. (2018) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-018-0036-9 · Cited by 1,445
Isolation of Extracellular Vesicles: General Methodologies and Latest Trends
Konoshenko, M. Y., Lekchnov, E. A., Vlassov, A. V., & Laktionov, P. P. (2018) · BioMed Research International · doi:10.1155/2018/8545347 · Cited by 1,243
Haraszti, R. A., Didiot, M., Sapp, E., Leszyk, J., Shaffer, S. A., Rockwell, H. E., Gao, F., Narain, N. R., DiFiglia, M., Kiebish, M. A., et al. (2016) · Journal of Extracellular Vesicles · doi:10.3402/jev.v5.32570 · Cited by 746
Metadata
- Authors
- Balaj, L., Atai, N. A., Chen, W., Mu, D., Tannous, B. A., Breakefield, X. O., Skog, J., & Maguire, C. A.
- Venue
- Scientific Reports
- Published
- 2015-05-19
- Volume
- 5
- Issue
- 1
- Pages
- 10266-10266
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W1888751811
- Cited by
- 189
- References
- 6
- FWCI
- 7.85
- Type
- journal article
- Language
- en
- Open access
- gold
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- Balaj, L., Atai, N. A., Chen, W., Mu, D., Tannous, B. A., Breakefield, X. O., Skog, J., & Maguire, C. A. (2015). Heparin affinity purification of extracellular vesicles. Scientific Reports. https://doi.org/10.1038/srep10266
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
