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Comparative evaluation of five extracellular vesicle isolation methods using proteomic profiling
| Autoři | |
|---|---|
| Rok publikování | 2026 |
| Druh | Článek v odborném periodiku |
| Časopis / Zdroj | AIMS Molecular Science |
| Fakulta / Pracoviště MU | |
| Citace | |
| www | https://www.aimspress.com/article/doi/10.3934/molsci.2026006 |
| Doi | https://doi.org/10.3934/molsci.2026006 |
| Klíčová slova | extracellular vesicles; isolation methods; comparison; mass spectrometry; proteomic analysis |
| Přiložené soubory | |
| Popis | Extracellular vesicles (EVs) research has gained a significant amount of attention in recent years. EVs are a heterogeneous group of different vesicles that vary in their origin, size, and function. The very nature of EVs, from their biogenesis to the contents of their cargo, offers many options for exploration. Despite being studied for a few decades now, there has not been an established standardized approach to isolation. However, the future use of EVs in clinical practice is conditioned by the optimization and standardization of isolation methods. In this study, we systematically compared common EV isolation techniques such as ultracentrifugation, precipitation, tangential flow filtration, and affinity-based methods using conditioned cell culture media as the source. Isolated EVs were analyzed using mass spectrometry to characterize their proteomic profiles, and the shared protein content was evaluated across datasets. The gene ontology enrichment was further assessed using three bioinformatics platforms. Among the tested methods, ultracentrifugation emerged as the most effective isolation method in our analysis, thus underscoring its importance among commonly used techniques. Despite its analytical robustness, this method is unsuitable for routine clinical workflows due to its complexity and time demands. Our findings highlight the need for a standardized, less strenuous EV isolation method for clinical applications. |