{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T14:55:07Z","timestamp":1785855307546,"version":"3.56.0"},"reference-count":15,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"vor","delay-in-days":21,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100004944","name":"Naresuan University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004944","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2026,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    This article presents a rigorous and reproducible protocol for the enrichment of plant\u2010derived nanovesicles (PDNVs) from the aquatic plant\n                    <jats:italic>Wolffia globosa<\/jats:italic>\n                    using differential ultracentrifugation combined with sucrose density gradient enrichment. The protocol is specifically optimized to address the technical challenges associated with PDNV enrichment from extremely small aquatic plants that require whole\u2010plant homogenization, a process that inherently increases contamination from intracellular organelles, soluble proteins, and chlorophyll. The workflow integrates gentle homogenization, sequential low\u2010 and medium\u2010speed centrifugation, sterile filtration, and an initial ultracentrifugation step to obtain crude PDNVs, followed by density\u2010based separation using a discontinuous sucrose gradient (8%\u201330% w\/v). This combined size\u2010 and density\u2010based strategy enables effective enrichment of PDNVs while substantially reducing non\u2010vesicular contaminants. Quality control and characterization are performed in accordance with MISEV2023 recommendations using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Gradient\u2010enriched PDNVs exhibit a predominantly nanoscale, homogeneous particle population with a mean diameter of approximately 105.3 nm and a particle concentration on the order of 10\n                    <jats:sup>10<\/jats:sup>\n                    particles\/ml. TEM analysis confirms well\u2010preserved membrane\u2010bound vesicles with characteristic round\u2010to\u2010cup\u2010shaped morphology and minimal background contamination, demonstrating a clear improvement over crude PDNV preparations. Quantitative normalization of PDNV yield to plant biomass and assessment of particle\u2010to\u2010protein ratios provide standardized metrics for evaluating preparation quality and reproducibility across independent preparations. Overall, this protocol provides a robust, scalable approach for obtaining enriched PDNVs prepared from\n                    <jats:italic>W.globosa<\/jats:italic>\n                    using commonly available laboratory equipment. The resulting PDNV preparations are suitable for downstream physicochemical characterization, molecular profiling, and functional studies, supporting their application in plant PDNV biology and emerging biomedical research. \u00a9 2026 Wiley Periodicals LLC.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Enrichment of plant\u2010derived nanovesicles (PDNVs) from the aquatic plant\n                    <jats:italic>W. globosa<\/jats:italic>\n                    by differential ultracentrifugation and sucrose density gradient\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70427","type":"journal-article","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T15:25:34Z","timestamp":1784733934000},"source":"Crossref","is-referenced-by-count":0,"title":["Enrichment and Characterization Protocol for Plant\u2010Derived Nanovesicles from the Aquatic Plant\n                    <i>Wolffia globosa<\/i>"],"prefix":"10.1002","volume":"6","author":[{"given":"Arunya","family":"Jiraviriyakul","sequence":"first","affiliation":[{"name":"Department of Medical Technology, Faculty of Allied Health Sciences Naresuan University  Phitsanulok Thailand"},{"name":"Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences Naresuan University  Phitsanulok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manatsanan","family":"Piamano","sequence":"additional","affiliation":[{"name":"Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences Naresuan University  Phitsanulok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Korawut","family":"Pimpa","sequence":"additional","affiliation":[{"name":"Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences Naresuan University  Phitsanulok 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