Analysis of small extracellular vesicles from dried blood spots
This protocol paper describes how to extract small extracellular vesicles (sEVs) from dried blood spots (DBS). The methodology is described in detail and offers further evidence that the extracted particles are sEVs using western blotting (anti-CD9, CD63 and CD81) and fluorescence nanoparticle track...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Medical Technology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmedt.2025.1494239/full |
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| author | Rikke Bæk Jenni Kathrine Sloth Mohammad Mehedi Hasan Getnet Midekessa Getnet Midekessa Malene Møller Jørgensen Malene Møller Jørgensen |
| author_facet | Rikke Bæk Jenni Kathrine Sloth Mohammad Mehedi Hasan Getnet Midekessa Getnet Midekessa Malene Møller Jørgensen Malene Møller Jørgensen |
| author_sort | Rikke Bæk |
| collection | DOAJ |
| description | This protocol paper describes how to extract small extracellular vesicles (sEVs) from dried blood spots (DBS). The methodology is described in detail and offers further evidence that the extracted particles are sEVs using western blotting (anti-CD9, CD63 and CD81) and fluorescence nanoparticle tracking analysis (fNTA). In addition, we present evidence that approximately 40% of the sEVs were recovered from DBS compared with EVs analyzed from plasma directly. The protocol proves to be robust, reliable and displays very interesting performances even after several weeks (up to 3 weeks) of storage of the DBS when analyzing the sEVs using protein microarray for the presence of the markers CD9, CD63, CD81, EpCAM, Flotilin-1, CD62E/P, CD142 and CD235a. These findings have important implications for using sEVs as future potential diagnostic tools by supporting the validity of less-invasive methods that can be implemented within vulnerable populations or in the field. |
| format | Article |
| id | doaj-art-1bc2751f47e74fc5a912f283115e666c |
| institution | Kabale University |
| issn | 2673-3129 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Medical Technology |
| spelling | doaj-art-1bc2751f47e74fc5a912f283115e666c2025-01-27T06:41:03ZengFrontiers Media S.A.Frontiers in Medical Technology2673-31292025-01-01710.3389/fmedt.2025.14942391494239Analysis of small extracellular vesicles from dried blood spotsRikke Bæk0Jenni Kathrine Sloth1Mohammad Mehedi Hasan2Getnet Midekessa3Getnet Midekessa4Malene Møller Jørgensen5Malene Møller Jørgensen6Department of Clinical Immunology, Aalborg University Hospital, Aalborg, DenmarkDepartment of Clinical Immunology, Aalborg University Hospital, Aalborg, DenmarkResearch Department of Maternal and Fetal Medicine, Elizabeth Garrett Anderson Institute for Women’s Health, University College London, London, United KingdomDepartment of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, EstoniaInstitute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu, EstoniaDepartment of Clinical Immunology, Aalborg University Hospital, Aalborg, DenmarkDepartment of Clinical Medicine, Aalborg University, Aalborg, DenmarkThis protocol paper describes how to extract small extracellular vesicles (sEVs) from dried blood spots (DBS). The methodology is described in detail and offers further evidence that the extracted particles are sEVs using western blotting (anti-CD9, CD63 and CD81) and fluorescence nanoparticle tracking analysis (fNTA). In addition, we present evidence that approximately 40% of the sEVs were recovered from DBS compared with EVs analyzed from plasma directly. The protocol proves to be robust, reliable and displays very interesting performances even after several weeks (up to 3 weeks) of storage of the DBS when analyzing the sEVs using protein microarray for the presence of the markers CD9, CD63, CD81, EpCAM, Flotilin-1, CD62E/P, CD142 and CD235a. These findings have important implications for using sEVs as future potential diagnostic tools by supporting the validity of less-invasive methods that can be implemented within vulnerable populations or in the field.https://www.frontiersin.org/articles/10.3389/fmedt.2025.1494239/fullsmall extracellular vesiclesdried blood spotsEV Arrayphenotypingwhole blood |
| spellingShingle | Rikke Bæk Jenni Kathrine Sloth Mohammad Mehedi Hasan Getnet Midekessa Getnet Midekessa Malene Møller Jørgensen Malene Møller Jørgensen Analysis of small extracellular vesicles from dried blood spots Frontiers in Medical Technology small extracellular vesicles dried blood spots EV Array phenotyping whole blood |
| title | Analysis of small extracellular vesicles from dried blood spots |
| title_full | Analysis of small extracellular vesicles from dried blood spots |
| title_fullStr | Analysis of small extracellular vesicles from dried blood spots |
| title_full_unstemmed | Analysis of small extracellular vesicles from dried blood spots |
| title_short | Analysis of small extracellular vesicles from dried blood spots |
| title_sort | analysis of small extracellular vesicles from dried blood spots |
| topic | small extracellular vesicles dried blood spots EV Array phenotyping whole blood |
| url | https://www.frontiersin.org/articles/10.3389/fmedt.2025.1494239/full |
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