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Extracellular vesicles (EVs) are promising biological nanovesicles which can be targeted by displaying binding proteins and are being developed as therapeutics. Currently, discovering EV targeting constructs is limited by low throughput and resource-intensive EV production and isolation. To accelerate discovery, we developed a screening pipeline to identify EV targeting constructs without requiring EV production. This approach is premised on the hypothesis that cell-cell interactions may predict some cell-EV interactions. Our\n                  <jats:underline>cell<\/jats:underline>\n                  binding as\n                  <jats:underline>say<\/jats:underline>\n                  (CELLISA) quantifies binding of a cell surface-displayed targeting protein to its cognate receptor on a target cell, employing a microscopy-based analysis pipeline. After validating the premise using existing T cell-targeting reagents, we develop CELLISA for either adherent or suspension EV producer cells. Finally, we use CELLISA to evaluate new binders and validate that hits mediate targeting and\/or delivery of genetic cargo to natural killer cells and T cells. CELLISA increased throughput &gt; 6-fold and decreased time by 40% compared to standard EV screens, and it identified a T-cell binder conferring efficient gene delivery. CELLISA is easily adaptable to other laboratories and can accelerate EV research.\n                <\/jats:p>","DOI":"10.64898\/2026.04.09.717595","type":"posted-content","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T01:30:18Z","timestamp":1776130218000},"source":"Crossref","is-referenced-by-count":0,"title":["CELLISA \u2013 a cell-cell binding assay for evaluation of nanovesicle targeting proteins"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2498-2630","authenticated-orcid":false,"given":"Taylor F.","family":"Gunnels","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA"},{"name":"Center for Synthetic Biology, Northwestern University, Evanston, IL, 60208, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6938-4813","authenticated-orcid":false,"given":"Jonathan D.","family":"Boucher","sequence":"additional","affiliation":[{"name":"Center for Synthetic Biology, Northwestern University, Evanston, IL, 60208, USA"},{"name":"Interdisciplinary Biological Sciences Training Program, Northwestern University, Evanston, IL, 60208, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5943-6018","authenticated-orcid":false,"given":"Yazeed S.","family":"Alroogi","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9362-6106","authenticated-orcid":false,"given":"Neha P.","family":"Kamat","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA"},{"name":"Center for Synthetic Biology, Northwestern University, Evanston, IL, 60208, USA"},{"name":"Interdisciplinary Biological Sciences Training Program, Northwestern University, Evanston, IL, 60208, USA"},{"name":"Chemistry of Life Processes Institute, Northwestern University, Evanston, IL, 60208, USA"},{"name":"Member, Robert H. 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