Cocoa Pulp as a Novel Source of Plant-Derived Exosome-Like Nanovesicles: Method Comparison and Physicochemical Characterization

Cocoa Pulp as a Novel Source of Plant-Derived Exosome-Like Nanovesicles: Method Comparison and Physicochemical Characterization | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Cocoa Pulp as a Novel Source of Plant-Derived Exosome-Like Nanovesicles: Method Comparison and Physicochemical Characterization Andrea Tapia-Aguayo, Stephanie Krüger, Ludger A. Wessjohann, María José Chica, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6989592/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Cocoa (Theobroma cacao L.) processing generates substantial agricultural residues, including pod husks and mucilaginous pulp, which remain largely underutilized. In this study, we report for the first time the isolation of plant-derived exosome-like nanovesicles (PENs) from cocoa pulp, a novel biological matrix with potential applications in sustainable nanobiotechnology. Two primary cell disruption methods, high-performance homogenization and cryogenic grinding, were compared for PEN recovery, followed by ultracentrifugation-based purification. The vesicles were characterized by cryo-transmission electron microscopy (Cryo-TEM), dynamic light scattering (DLS), and bicinchoninic acid (BCA) protein assays. PENs isolated via cryogenic processing exhibited more uniform size distributions (111.9 ± 2.4 nm) and spherical morphology compared to those obtained by homogenization (120.2 ± 5.5 nm). While homogenization yielded higher protein concentrations, it also resulted in greater vesicle heterogeneity. No vesicles were detected from cocoa pod husks under the tested conditions. These findings establish cocoa pulp as a promising and sustainable source of PENs and demonstrate how primary recovery methods critically influence vesicle quality. This methodological contribution lays the groundwork for future studies on PEN bioactivity, cargo profiling, and functional applications. Biological sciences/Biochemistry Biological sciences/Biological techniques Biological sciences/Biotechnology Physical sciences/Nanoscience and technology Biological sciences/Plant sciences Plant-derived Exosome-like Nanovesicles (PENs) Cocoa residues Primary Recovery Operations High-performance Homogenizing Cryogenic Grinding Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6989592","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":477319557,"identity":"2ed97c38-7fed-4c4a-8831-8ea7a9a0c60c","order_by":0,"name":"Andrea Tapia-Aguayo","email":"","orcid":"","institution":"Monterrey Institute of Technology","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"","lastName":"Tapia-Aguayo","suffix":""},{"id":477319558,"identity":"76ab81f4-01db-4719-96a9-e260f82f6566","order_by":1,"name":"Stephanie Krüger","email":"","orcid":"","institution":"Martin-Luther-University","correspondingAuthor":false,"prefix":"","firstName":"Stephanie","middleName":"","lastName":"Krüger","suffix":""},{"id":477319559,"identity":"51fe7ca0-04e6-47d0-8a83-6654efef23c4","order_by":2,"name":"Ludger A. 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In this study, we report for the first time the isolation of plant-derived exosome-like nanovesicles (PENs) from cocoa pulp, a novel biological matrix with potential applications in sustainable nanobiotechnology. Two primary cell disruption methods, high-performance homogenization and cryogenic grinding, were compared for PEN recovery, followed by ultracentrifugation-based purification. The vesicles were characterized by cryo-transmission electron microscopy (Cryo-TEM), dynamic light scattering (DLS), and bicinchoninic acid (BCA) protein assays. PENs isolated via cryogenic processing exhibited more uniform size distributions (111.9 ± 2.4 nm) and spherical morphology compared to those obtained by homogenization (120.2 ± 5.5 nm). While homogenization yielded higher protein concentrations, it also resulted in greater vesicle heterogeneity. No vesicles were detected from cocoa pod husks under the tested conditions. 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