Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions
Abstract Separating extracellular vesicles (EV) from blood plasma is challenging and complicates their biological understanding and biomarker development. In this study, we fractionate blood plasma by combining size‐exclusion chromatography (SEC) and OptiPrep density gradient centrifugation to study...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-08-01
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| Series: | Journal of Extracellular Vesicles |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jev2.12122 |
| _version_ | 1819349529879642112 |
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| author | Glenn Vergauwen Joeri Tulkens Cláudio Pinheiro Francisco Avila Cobos Sándor Dedeyne Marie‐Angélique De Scheerder Linos Vandekerckhove Francis Impens Ilkka Miinalainen Geert Braems Kris Gevaert Pieter Mestdagh Jo Vandesompele Hannelore Denys Olivier De Wever An Hendrix |
| author_facet | Glenn Vergauwen Joeri Tulkens Cláudio Pinheiro Francisco Avila Cobos Sándor Dedeyne Marie‐Angélique De Scheerder Linos Vandekerckhove Francis Impens Ilkka Miinalainen Geert Braems Kris Gevaert Pieter Mestdagh Jo Vandesompele Hannelore Denys Olivier De Wever An Hendrix |
| author_sort | Glenn Vergauwen |
| collection | DOAJ |
| description | Abstract Separating extracellular vesicles (EV) from blood plasma is challenging and complicates their biological understanding and biomarker development. In this study, we fractionate blood plasma by combining size‐exclusion chromatography (SEC) and OptiPrep density gradient centrifugation to study clinical context‐dependent and time‐dependent variations in the biomolecular landscape of systemically circulating EV. Using pooled blood plasma samples from breast cancer patients, we first demonstrate the technical repeatability of blood plasma fractionation. Using serial blood plasma samples from HIV and ovarian cancer patients (n = 10) we next show that EV carry a clinical context‐dependent and/or time‐dependent protein and small RNA composition, including miRNA and tRNA. In addition, differential analysis of blood plasma fractions provides a catalogue of putative proteins not associated with systemically circulating EV. In conclusion, the implementation of blood plasma fractionation allows to advance the biological understanding and biomarker development of systemically circulating EV. |
| first_indexed | 2024-12-24T19:01:59Z |
| format | Article |
| id | doaj.art-83a21f9d3adf44778a5e98596e49c8c9 |
| institution | Directory Open Access Journal |
| issn | 2001-3078 |
| language | English |
| last_indexed | 2024-12-24T19:01:59Z |
| publishDate | 2021-08-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Vesicles |
| spelling | doaj.art-83a21f9d3adf44778a5e98596e49c8c92022-12-21T16:43:12ZengWileyJournal of Extracellular Vesicles2001-30782021-08-011010n/an/a10.1002/jev2.12122Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditionsGlenn Vergauwen0Joeri Tulkens1Cláudio Pinheiro2Francisco Avila Cobos3Sándor Dedeyne4Marie‐Angélique De Scheerder5Linos Vandekerckhove6Francis Impens7Ilkka Miinalainen8Geert Braems9Kris Gevaert10Pieter Mestdagh11Jo Vandesompele12Hannelore Denys13Olivier De Wever14An Hendrix15Department of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumCancer Research Institute Ghent Ghent BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Internal Medicine and Pediatrics HIV Cure Research Center Ghent University Hospital Ghent BelgiumDepartment of Internal Medicine and Pediatrics HIV Cure Research Center Ghent University Hospital Ghent BelgiumVIB Center for Medical Biotechnology Ghent BelgiumBiocenter Oulu University of Oulu Oulu FinlandCancer Research Institute Ghent Ghent BelgiumVIB Center for Medical Biotechnology Ghent BelgiumCancer Research Institute Ghent Ghent BelgiumCancer Research Institute Ghent Ghent BelgiumCancer Research Institute Ghent Ghent BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumDepartment of Human Structure and Repair Laboratory of Experimental Cancer Research Ghent University Ghent BelgiumAbstract Separating extracellular vesicles (EV) from blood plasma is challenging and complicates their biological understanding and biomarker development. In this study, we fractionate blood plasma by combining size‐exclusion chromatography (SEC) and OptiPrep density gradient centrifugation to study clinical context‐dependent and time‐dependent variations in the biomolecular landscape of systemically circulating EV. Using pooled blood plasma samples from breast cancer patients, we first demonstrate the technical repeatability of blood plasma fractionation. Using serial blood plasma samples from HIV and ovarian cancer patients (n = 10) we next show that EV carry a clinical context‐dependent and/or time‐dependent protein and small RNA composition, including miRNA and tRNA. In addition, differential analysis of blood plasma fractions provides a catalogue of putative proteins not associated with systemically circulating EV. In conclusion, the implementation of blood plasma fractionation allows to advance the biological understanding and biomarker development of systemically circulating EV.https://doi.org/10.1002/jev2.12122biomarkersbloodcoronaexosomesextracellular vesiclesisolation |
| spellingShingle | Glenn Vergauwen Joeri Tulkens Cláudio Pinheiro Francisco Avila Cobos Sándor Dedeyne Marie‐Angélique De Scheerder Linos Vandekerckhove Francis Impens Ilkka Miinalainen Geert Braems Kris Gevaert Pieter Mestdagh Jo Vandesompele Hannelore Denys Olivier De Wever An Hendrix Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions Journal of Extracellular Vesicles biomarkers blood corona exosomes extracellular vesicles isolation |
| title | Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| title_full | Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| title_fullStr | Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| title_full_unstemmed | Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| title_short | Robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| title_sort | robust sequential biophysical fractionation of blood plasma to study variations in the biomolecular landscape of systemically circulating extracellular vesicles across clinical conditions |
| topic | biomarkers blood corona exosomes extracellular vesicles isolation |
| url | https://doi.org/10.1002/jev2.12122 |
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