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The enrichment of tissue\u2010specific EVs (TS\u2010EVs) from human biofluids can be challenging due to technical and methodological limitations. Here, we use single and sequential immunoaffinity capture workflows to enrich antigen\u2010specific EVs circulating in human blood. We demonstrate the specificity, efficiency, and consistency of our approach for enriching blood plasma EV subpopulations from the nervous system, alveolar cells and hepatocytes. The enriched subpopulations are characterized by canonical EV features and markers, as well as co\u2010localization of tissue\u2010specific and general markers on the surface. We provide a validated workflow to derive multiple EV subpopulations from circulation, leveraging their promise as informative biomarkers of tissue status and enabling liquid biopsy and biomarker discovery.<\/jats:p>","DOI":"10.1002\/jex2.70171","type":"journal-article","created":{"date-parts":[[2026,7,14]],"date-time":"2026-07-14T01:22:27Z","timestamp":1783992147000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Immunoaffinity\u2010Based Protocol to Enrich Nervous System Cell\u2010, Lung Alveolar Cell\u2010, and Hepatocyte\u2010Derived Extracellular Vesicles From Human Plasma"],"prefix":"10.1002","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5248-3014","authenticated-orcid":false,"given":"Biancamaria","family":"Pierri","sequence":"first","affiliation":[{"name":"Department of Environmental Health Sciences Mailman School of Public Health Columbia University  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela L.","family":"Jackson","sequence":"additional","affiliation":[{"name":"Department of Environmental Health Sciences Mailman School of Public Health Columbia University  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0392-268X","authenticated-orcid":false,"given":"Olesia","family":"Gololobova","sequence":"additional","affiliation":[{"name":"Molecular and Comparative Pathobiology and Neurology The Johns Hopkins University School of Medicine  Baltimore Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Environmental Health Sciences Mailman School of Public Health Columbia University  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erez","family":"Eitan","sequence":"additional","affiliation":[{"name":"NeuroDex Inc.  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