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Herein, we present a single\u2010EV and particle (siEVP) protein and RNA assay (\n                    <jats:sup>siEVP<\/jats:sup>\n                    PRA) to simultaneously detect mRNAs, miRNAs, and proteins in subpopulations of EVs and LPs. The\n                    <jats:sup>siEVP<\/jats:sup>\n                    PRA immobilizes and sorts particles via positive immunoselection onto micropatterns and focuses biomolecular signals in situ. By detecting EVPs at a single\u2010particle resolution, the\n                    <jats:sup>siEVP<\/jats:sup>\n                    PRA outperformed the sensitivities of bulk\u2010analysis benchmark assays for RNA and protein. To assess the specificity of RNA detection in complex biofluids, EVs from various glioma cell lines were processed with small RNA sequencing, whereby two mRNAs and two miRNAs associated with glioblastoma multiforme (GBM) were chosen for cross\u2010validation. Despite the presence of single\u2010EV\u2010LP co\u2010isolates in serum, the\n                    <jats:sup>siEVP<\/jats:sup>\n                    PRA detected GBM\u2010associated vesicular RNA profiles in GBM patient siEVPs. The\n                    <jats:sup>siEVP<\/jats:sup>\n                    PRA effectively examines intravesicular, intervesicular, and interparticle heterogeneity with diagnostic promise.\n                  <\/jats:p>","DOI":"10.1002\/jev2.12369","type":"journal-article","created":{"date-parts":[[2023,11,1]],"date-time":"2023-11-01T02:49:50Z","timestamp":1698806990000},"source":"Crossref","is-referenced-by-count":31,"title":["Engineering a tunable micropattern\u2010array assay to sort single extracellular vesicles and particles to detect RNA and protein in situ"],"prefix":"10.1002","volume":"12","author":[{"given":"Jingjing","family":"Zhang","sequence":"first","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5633-5887","authenticated-orcid":false,"given":"Xilal Y.","family":"Rima","sequence":"additional","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Wang","sequence":"additional","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1811-6194","authenticated-orcid":false,"given":"Luong T. H.","family":"Nguyen","sequence":"additional","affiliation":[{"name":"William G. 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Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paola Loreto","family":"Palacio","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Neurology Nationwide Children's Hospital  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim Truc","family":"Nguyen","sequence":"additional","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karunya","family":"Albert","sequence":"additional","affiliation":[{"name":"William G. 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Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Li","sequence":"additional","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob","family":"Doon\u2010Ralls","sequence":"additional","affiliation":[{"name":"William G. Lowrie Department of Chemical and Biomolecular Engineering The Ohio State University  Columbus Ohio USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin L.","family":"Hisey","sequence":"additional","affiliation":[{"name":"William G. 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