The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.
Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research
Open accessAbstract
Identification and validation of extracellular vesicle (EV)-associated biomarkers requires robust isolation and characterization protocols. We assessed the impact of some commonly implemented pre-analytical, analytical and post-analytical variables in EV research. Centrifugal filters with different membrane types and pore sizes are used to reduce large volume biofluids prior to EV isolation or to concentrate EVs. We compared five commonly reported filters for their efficiency when using plasma, urine and EV-spiked PBS. Regenerated cellulose membranes with pore size of 10 kDa recovered EVs the most efficient. Less than 40% recovery was achieved with other filters. Next, we analyzed the effect of the type of protein assays to measure EV protein in colorimetric and fluorometric kits. The fluorometric assay Qubit measured low concentration EV and BSA samples the most accurately with the lowest variation among technical and biological replicates. Lastly, we quantified Optiprep remnants in EV samples from density gradient ultracentrifugation and demonstrate that size-exclusion chromatography efficiently removes Optiprep from EVs. In conclusion, choice of centrifugal filters and protein assays confound EV analysis and should be carefully considered to increase efficiency towards biomarker discovery. SEC-based removal of Optiprep remnants from EVs can be considered for downstream applications.
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Cites
Glypican-1 identifies cancer exosomes and detects early pancreatic cancer
Melo, S. A., Luecke, L. B., Kahlert, C., Fernandez, A. F., Gammon, S. T., Kaye, J., LeBleu, V. S., Mittendorf, E. A., Weitz, J., Rahbari, N., et al. (2015) · Nature · doi:10.1038/nature14581 · Cited by 2,880
EV-TRACK: transparent reporting and centralizing knowledge in extracellular vesicle research
EV-TRACK Consortium, Van Deun, J., Mestdagh, P., Agostinis, P., Akay, Ö., Anand, S., Anckaert, J., Martinez, Z. A., Baetens, T., Beghein, E., et al. (2017) · Nature Methods · doi:10.1038/nmeth.4185 · Cited by 1,284
Self-citationActivity-associated miRNA are packaged in Map1b-enriched exosomes released from depolarized neurons
Goldie, B. J., Dun, M. D., Lin, M., Smith, N. D., Verrills, N. M., Dayas, C. V., & Cairns, M. J. (2014) · Nucleic Acids Research · doi:10.1093/nar/gku594 · Cited by 318
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
Self-citationZhang, Y., Bi, J., Huang, J., Tang, Y., Du, S., & Li, P. (2020) · International Journal of Nanomedicine · doi:10.2147/ijn.s264498 · Cited by 1,323
Meng, W., He, C., Hao, Y., Wang, L., Li, L., & Zhu, G. (2020) · Drug Delivery · doi:10.1080/10717544.2020.1748758 · Cited by 592
Erdbrügger, U., Blijdorp, C. J., Bijnsdorp, I. V., Borràs, F. E., Burger, D., Bussolati, B., Byrd, J. B., Clayton, A., Dear, J. W., Falcón‐Pérez, J. M., et al. (2021) · Journal of Extracellular Vesicles · doi:10.1002/jev2.12093 · Cited by 404
Metadata
- Authors
- Vergauwen, G., Dhondt, B., Van Deun, J., De Smedt, E., Berx, G., Timmerman, E., Gevaert, K., Miinalainen, I., Cocquyt, V., Braems, G., Van den Broecke, R., Denys, H., De Wever, O., & Hendrix, A.
- Venue
- Scientific Reports
- Published
- 2017-06-02
- Volume
- 7
- Issue
- 1
- Pages
- 2704-2704
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W2619136320
- Cited by
- 270
- References
- 3
- FWCI
- 6.89
- Type
- journal article
- Language
- en
- Open access
- gold
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- Vergauwen, G., Dhondt, B., Van Deun, J., De Smedt, E., Berx, G., Timmerman, E., Gevaert, K., Miinalainen, I., Cocquyt, V., Braems, G., Van den Broecke, R., Denys, H., De Wever, O., & Hendrix, A. (2017). Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research. Scientific Reports. https://doi.org/10.1038/s41598-017-02599-y
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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).
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