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Isolation of Citrus lemon extracellular vesicles: Development and process control using capillary electrophoresis
Abstract
A new and scalable method for the isolation of extracellular vesicles (EV) from Citrus lemon juice samples was developed. The methodology included preliminary preconcentration of the sample using ultrafiltration (UF) followed by size-exclusion chromatography (SEC) purification and final preconcentration of the eluates. Transmission electron microscopy and proteomic analysis showed that isolates contained exosome-like vesicles, exocyst-positive organelle (EXPO), and microvesicles. The efficiency of certain isolation steps was evaluated with total protein content assay (bicinchoninic acid assay, BCA), nanoparticles tracking analysis (NTA), and capillary electrophoresis (CE). A good correlation between CE, BCA, and NTA results was shown. The application of CE enabled the detection of soluble contaminants, macromolecular aggregates, and vesicles’ heterogeneity. The fluorescent staining of encapsulated nucleic acids was proposed for the identity confirmation of EV detected in CE. The study demonstrates the CE as a comprehensive tool for monitoring of the EV isolation process.
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Authors (10)
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Aleksandra Steć
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Martyna Chodkowska
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Joanna Kasprzyk-Pochopień
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Przemyslaw Mielczarek
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Wojciech Piekoszewski
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Bogdan Lewczuk
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Agata Płoska
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Bartosz Wielgomas
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Szymon Dziomba
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Details
- Category:
- Articles
- Type:
- artykuły w czasopismach
- Published in:
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FOOD CHEMISTRY
no. 424,
ISSN: 0308-8146 - Language:
- English
- Publication year:
- 2023
- Bibliographic description:
- Steć A., Chodkowska M., Kasprzyk-Pochopień J., Mielczarek P., Piekoszewski W., Lewczuk B., Płoska A., Kalinowski L., Wielgomas B., Dziomba S.: Isolation of Citrus lemon extracellular vesicles: Development and process control using capillary electrophoresis// FOOD CHEMISTRY -Vol. 424, (2023), s.136333-
- DOI:
- Digital Object Identifier (open in new tab) 10.1016/j.foodchem.2023.136333
- Sources of funding:
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- Polish Ministry of Education and Science, grants no. 10/E-389/SPUB/SP/2020 and DIR/WK/2017/01
- Verified by:
- Gdańsk University of Technology
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