Generalizability of Enzyme‐Based Isolation Approach for Extracellular Vesicles From Traditional Medicinal Plants

Extracellular vesicles (EVs) from plants, particularly traditional medicinal plants, have garnered therapeutic interest. However, therapeutic utility requires establishing effective extraction and purification protocols for manufacturing. Here, we systemically optimized an enzyme-based EV extraction method and demonstrated the generalizability with different parts from seventeen medicinal plants, fresh, frozen or dried. The enzymatic pulping process (henceforth denoted as Phyto-EVpure) of fresh ginger, <i>Morus alba</i> leaves and <i>Isatis indigotica Fort</i>. roots improved EV yield and purity over grinding. Ginger EVs processed via Phyto-EVpure preserved bioactivities and conferred hepatoprotection in a non-alcoholic fatty liver disease model in vitro and in vivo. Phyto-EVpure efficiently isolated EVs from leaf, rhizome and root stored fresh, frozen or dried. Phyto-EVpure enabled us to demonstrate that EVs from phloem of previously intractable dried vine samples exhibited greater antitumour activities than xylem in vitro. Collectively, we provided a manufacturing-friendly protocol for medicinal plant EV isolation and demonstrated the generalizability of Phyto-EVpure for efficient isolation of EVs from a wide spectrum of traditional medicinal plants under different conditions.