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METHODS article

Front. Med. Technol.
Sec. Diagnostic and Therapeutic Devices
Volume 7 - 2025 | doi: 10.3389/fmedt.2025.1494239

Analysis of small extracellular vesicles from dried blood spots

Provisionally accepted
Rikke Bæk Rikke Bæk 1Jenni Katrine Sloth Jenni Katrine Sloth 1Mohammad Mehedi Hasan Mohammad Mehedi Hasan 2Gtnet Midekessa Gtnet Midekessa 3,4Malene Møller Jørgensen Malene Møller Jørgensen 1,5*
  • 1 Department of Clinical Immunology, Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark
  • 2 University College London, Elizabeth Garrett Anderson Institute for Women’s Health, Research Department of maternal and fetal medicine, London, United Kingdom
  • 3 Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, Faculty of Medicine, University of Tartu, Tartu, Tartu County, Estonia
  • 4 Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Tartu, Tartu County, Estonia
  • 5 Department of Clinical Medicine, Aalborg University, Aalborg, Denmark

The final, formatted version of the article will be published soon.

    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.

    Keywords: Small extracellular vesicles, Dried blood spots, EV Array, phenotyping, whole blood

    Received: 10 Sep 2024; Accepted: 14 Jan 2025.

    Copyright: © 2025 Bæk, Sloth, Hasan, Midekessa and Jørgensen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Malene Møller Jørgensen, Department of Clinical Immunology, Department of Clinical Immunology, Aalborg University Hospital, Aalborg, 9000, Denmark

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.