Identity confirmation of extracellular vesicles by capillary electrophoresis using non-specific dyes

[Display omitted] •Non-specific dyes were used for extracellular vesicle (EV) staining.•The approach was used for the confirmation of EV identity.•Separation and detection of plant and bacteria EV subpopulations was possible. Extracellular vesicles (EVs) are membrane-enveloped nanostructures secrete...

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Bibliographic Details
Published in:Microchemical journal 2024-12, Vol.207, p.112288, Article 112288
Main Authors: Steć, Aleksandra, Klocek, Kamil, Czyrski, Grzegorz S., Heinz, Andrea, Dziomba, Szymon
Format: Article
Language:English
Subjects:
Capillary electrophoresis
Citrus aurantifolia
Citrus limon
Escherichia coli
Extracellular vesicles
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Summary:[Display omitted] •Non-specific dyes were used for extracellular vesicle (EV) staining.•The approach was used for the confirmation of EV identity.•Separation and detection of plant and bacteria EV subpopulations was possible. Extracellular vesicles (EVs) are membrane-enveloped nanostructures secreted by prokaryotic and eukaryotic cells. Carrying a variety of essential components, EVs are involved in numerous biological processes including intercellular communication and immunological response. The characterization of extracellular vesicles with capillary electrophoresis (CE) faces several challenges. Due to the limited availability of commercial standards, the identity confirmation of the EV signal is among these issues. In the presented work, a method for identity confirmation of EVs by CE was developed. Identification was based on the application of non-specific dyes and fluorescence detection, and EVs obtained from Escherichia coli and citrus fruits (Citrus limon and Citrus aurantifolia) were used. It was found that the presented methodology effectively detects the cargo carried by EVs (including nucleic acids, proteins, and lipids) and enables the staining of subpopulations separated by CE. The strategy holds promise to be utilized for EV heterogeneity assessment.
ISSN:0026-265X
DOI:10.1016/j.microc.2024.112288