{
  "abstract": "The antioxidant activity of extracellular vesicles (EVs) and nanovesicles are increasingly linked to their physiological and therapeutic functions, yet its accurate quantification remains limited by low EV yield, co‐isolated contaminants and the lack of standardized methodologies. Here, we establish optimized microplate‐based protocols for ABTS and ORAC assays specifically adapted to highly purified EVs and nanovesicles obtained by ultracentrifugation followed by size‐exclusion chromatography (UC + SEC). By minimizing sample input, implementing protein‐based normalization (TEAC/g EV protein), and introducing assay‐specific controls—including a self‐blank correction for ORAC—we achieve sensitive and reproducible measurements across plant‐, human plasma‐ and bacterial‐derived EVs and nanovesicles. Our results demonstrate that conventional isolation methods substantially overestimate antioxidant activity due to non‐vesicular contaminants, underscoring the need for rigorous purification. Using a detergent‐based disruption strategy compatible with ABTS, we further distinguish surface‐associated from cargo‐derived antioxidant activity, revealing a two‐tier functional organization of EVs. Notably, we report for the first time measurable antioxidant activity in bacterial extracellular vesicles (BEVs). Application of these methods to a human dietary intervention shows that EV antioxidant capacity is dynamically modulated in vivo. Collectively, this work provides a standardized and broadly applicable framework for the reliable assessment of EV and nanovesicles antioxidant properties across biological kingdoms.",
  "abstract_shown": true,
  "affiliations": [
    "Laboratory of Epigenetics of Lipid Metabolism Instituto Madrileño de Estudios Avanzados (IMDEA)‐Nutrición, CEI UAM+CSIC Madrid Spain",
    "Bioactivity and Nutritional Immunology Group (BIOINUT), Faculty of Health Science Universidad Internacional de Valencia (VIU) Valencia Spain",
    "Department of Nutrition and Metabolism, Institute of Food Science, Technology and Nutrition Spanish National Research Council, CSIC‐ICTAN Madrid Spain",
    "Department of Pharmaceutical Sciences Università Degli Studi di Milano Milan Italy",
    "Laboratory of Functional Foods Instituto Madrileño de Estudios Avanzados (IMDEA)‐Nutrición, CEI UAM+CSIC Madrid Spain",
    "Department of Molecular Medicine University of Padova Padova Italy",
    "Facultad de Ciencias de la Salud Universidad Tecnológica Atlántico Mediterráneo ‐ UTAMED Málaga Spain",
    "Laboratory of Food & Health, Research Group on Quality, Safety, and Bioactivity of Plant Foods, CEBAS‐CSIC Campus Universitario de Espinardo Murcia Spain",
    "Nutraceuticals and Neuroendocrine and Neurodegenerative Diseases Group (NUENE), Institute of Biomedicine. Faculty of Environmental Sciences and Biochemistry University of Castilla‐La Mancha Toledo Spain",
    "Plant Molecular Genetics Department, National Centre for Biotechnology (CNB) Consejo Superior de Investigaciones Cientificas (CSIC) Madrid Spain",
    "Consorcio CIBER de la Fisiopatología de la Obesidad y Nutrición (CIBERObn) Instituto de Salud Carlos III (ISCIII) Madrid Spain"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 16,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Juan Antonio Nieto",
      "orcid": "0000-0002-9266-0190"
    },
    {
      "affs": [
        0,
        2
      ],
      "name": "María‐Carmen López de las Hazas"
    },
    {
      "affs": [
        0
      ],
      "name": "Victoria Martín‐Santiago",
      "orcid": "0009-0004-5161-8895"
    },
    {
      "affs": [
        0
      ],
      "name": "Laura Rubio‐Silvan"
    },
    {
      "affs": [
        0
      ],
      "name": "Carmen Mazarío‐Gárgoles"
    },
    {
      "affs": [
        0
      ],
      "name": "Ludovica Attanasio"
    },
    {
      "affs": [
        3,
        0
      ],
      "name": "Benedetta Mercuriali",
      "orcid": "0009-0005-0296-9194"
    },
    {
      "affs": [
        4
      ],
      "name": "João Tomé‐Carneiro",
      "orcid": "0000-0002-3597-9234"
    },
    {
      "affs": [
        0,
        5
      ],
      "name": "Giulia Bernabè"
    },
    {
      "affs": [
        0,
        6
      ],
      "name": "Andrea del Saz‐Lara"
    },
    {
      "affs": [
        4
      ],
      "name": "Carmen Crespo",
      "orcid": "0000-0003-0546-2143"
    },
    {
      "affs": [
        7
      ],
      "name": "María Ángeles Ávila‐Gálvez",
      "orcid": "0000-0003-2513-1026"
    },
    {
      "affs": [
        8
      ],
      "name": "Emma Burgos‐Ramos",
      "orcid": "0000-0002-6942-3023"
    },
    {
      "affs": [
        7
      ],
      "name": "Juan Carlos Espín",
      "orcid": "0000-0002-1068-8692"
    },
    {
      "affs": [
        9
      ],
      "name": "Andrea Chini",
      "orcid": "0000-0001-5841-0254"
    },
    {
      "affs": [
        0,
        10
      ],
      "name": "Alberto Dávalos",
      "orcid": "0000-0001-5709-6443"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/9kavx8vkpr77n.bib",
  "cited_by": 0,
  "doi": "10.1002/jev2.70392",
  "fulltext_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13730445/?tool=EBI",
  "id": "9kavx8vkpr77n",
  "issns": [
    "2001-3078"
  ],
  "landing_url": "https://europepmc.org/article/PMC/PMC13730445",
  "language": "eng",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Research Article"
    ],
    [
      "openalex",
      "1312|Extracellular vesicles in disease"
    ]
  ],
  "openalex": "W7225200019",
  "pdf_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13730445/pdf/?tool=EBI",
  "pmcid": "PMC13730445",
  "pmid": "42976256",
  "published": "2026-11-01",
  "publisher": "Wiley",
  "ris": "https://research.draconitic.com/paper/9kavx8vkpr77n.ris",
  "sources": [
    {
      "first_seen": "2026-11-04T15:20:46Z",
      "native_id": "PMC:PMC13730445",
      "source": "europepmc",
      "url": "https://europepmc.org/article/PMC/PMC13730445"
    },
    {
      "first_seen": "2026-11-07T12:23:00Z",
      "native_id": "PMC13730445",
      "source": "pmc",
      "url": "https://doi.org/10.1002/jev2.70392"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Extracellular vesicles in disease",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Standardized Microplate‐Based Assays for Quantifying Antioxidant Activity in Extracellular Vesicles and Nanovesicles Across Biological Kingdoms",
  "title_text": "Standardized Microplate‐Based Assays for Quantifying Antioxidant Activity in Extracellular Vesicles and Nanovesicles Across Biological Kingdoms",
  "type": "peer-reviewed",
  "updated_at": "2026-11-07T12:23:00Z",
  "url": "https://research.draconitic.com/paper/9kavx8vkpr77n",
  "venue": "Journal of extracellular vesicles",
  "versions": []
}