{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:35:53Z","timestamp":1787027753000,"version":"build-2736575974"},"reference-count":39,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:00:00Z","timestamp":1760400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011033","name":"Spanish State Research Agency","doi-asserted-by":"publisher","award":["PID2019-106625RB-I00"],"award-info":[{"award-number":["PID2019-106625RB-I00"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011033","name":"Spanish State Research Agency","doi-asserted-by":"publisher","award":["PID2022-136453OB-I00"],"award-info":[{"award-number":["PID2022-136453OB-I00"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation","doi-asserted-by":"publisher","award":["PDC2022-133363-I00"],"award-info":[{"award-number":["PDC2022-133363-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ajuntament de Barcelona","award":["24S05876-001"],"award-info":[{"award-number":["24S05876-001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Exosomes are nanoscale extracellular vesicles that carry valuable biomolecular information. However, their characterization still depends on complex and costly techniques such as flow cytometry. In this study, a paper-based Vertical Flow Assay (VFA) specifically designed for the detection and profiling of exosomes derived from metastatic breast cancer cell lines is presented. The assay operates in an ELISA-like format, targeting exosomal surface proteins (CD9, CD63, CD81, and EGFR1) with specific antibodies and a secondary antibody conjugated to alkaline phosphatase. Upon reaction with the NBT\/BCIP substrate, an insoluble indigo precipitate forms on the nitrocellulose membrane, generating a visual signal that can be further quantified by smartphone imaging. The VFA was optimized for membrane type, pore size, and blocking agents, reaching a detection limit of ~6 \u00d7 107 exosomes \u00b5L\u22121 in less than 20 min. Comparative studies with bead-based flow cytometry confirmed consistent biomarker expression profiles, demonstrating the reliability of the method. By enabling exosome biomarker profiling in a simplified and low-cost format, this approach provides a promising alternative to flow cytometry and other applications required for exosome characterization.<\/jats:p>","DOI":"10.3390\/bios15100694","type":"journal-article","created":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T10:12:29Z","timestamp":1760523149000},"page":"694","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Exosome Biomarker Profiling Using a Paper-Based Vertical Flow Assay"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9990-148X","authenticated-orcid":false,"given":"Arnau","family":"Pallar\u00e8s-Rusi\u00f1ol","sequence":"first","affiliation":[{"name":"Grup de Sensors i Biosensors, Departament de Qu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"},{"name":"Biosensing and Bioanalysis Group, Institute of Biotechnology and Biomedicine, Universitat Aut\u00f2noma de Barcelona, Bellaterra 08193, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5321-3841","authenticated-orcid":false,"given":"Jennifer","family":"Marf\u00e0","sequence":"additional","affiliation":[{"name":"Grup de Sensors i Biosensors, Departament de Qu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"},{"name":"Biosensing and Bioanalysis Group, Institute of Biotechnology and Biomedicine, Universitat Aut\u00f2noma de Barcelona, Bellaterra 08193, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3656-2908","authenticated-orcid":false,"given":"Rosanna","family":"Rossi","sequence":"additional","affiliation":[{"name":"Grup de Sensors i Biosensors, Departament de Qu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"},{"name":"Biosensing and Bioanalysis Group, Institute of Biotechnology and Biomedicine, Universitat Aut\u00f2noma de Barcelona, Bellaterra 08193, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1846-0043","authenticated-orcid":false,"given":"Merc\u00e8","family":"Mart\u00ed","sequence":"additional","affiliation":[{"name":"Biosensing and Bioanalysis Group, Institute of Biotechnology and Biomedicine, Universitat Aut\u00f2noma de Barcelona, Bellaterra 08193, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5266-7873","authenticated-orcid":false,"given":"Mar\u00eda Isabel","family":"Pividori","sequence":"additional","affiliation":[{"name":"Grup de Sensors i Biosensors, Departament de Qu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"},{"name":"Biosensing and Bioanalysis Group, Institute of Biotechnology and Biomedicine, Universitat Aut\u00f2noma de Barcelona, Bellaterra 08193, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Moura, S.L., Pallar\u00e8s-Rusi\u00f1ol, A., Sappia, L., Mart\u00ed, M., and Pividori, M.I. 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