Analysis of small extracellular vesicles from dried blood spots.

IF 3.8 Q3 ENGINEERING, BIOMEDICAL Frontiers in medical technology Pub Date : 2025-01-27 eCollection Date: 2025-01-01 DOI:10.3389/fmedt.2025.1494239

Rikke Bæk, Jenni Kathrine Sloth, Mohammad Mehedi Hasan, Getnet Midekessa, Malene Møller Jørgensen

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Abstract

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.

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干血斑细胞外小泡分析。

本文描述了如何从干血斑（DBS）中提取小细胞外囊泡（sev）。详细描述了方法，并提供了进一步的证据，证明提取的颗粒是sev，使用western blotting（抗cd9， CD63和CD81）和荧光纳米颗粒跟踪分析（fNTA）。此外，我们提供的证据表明，与直接从血浆中分析的ev相比，大约40%的sev是从DBS中回收的。该方案被证明是稳健可靠的，并且在使用蛋白质微阵列分析sev时，即使在DBS存储几周（最多3周）后也显示出非常有趣的性能，用于标记CD9， CD63, CD81, EpCAM, Flotilin-1, CD62E/P， CD142和CD235a。这些发现对sev作为未来潜在的诊断工具具有重要意义，因为它支持了可在脆弱人群或现场实施的低侵入性方法的有效性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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