Isolation of extracellular vesicles from citrus limon fruit through membrane-based techniques.

The project is aimed at the development of a membrane-based protocol for the isolation of plant-derived extracellular vesicles. In particular, citrus limon fruit is selected as source material, due to its beneficial phyto-properties, availability and cheapness. In the chosen strategy the focus is placed upon the development of a protocol potentially suitable for large-scale operations, conversely to differential ultracentrifugation method, the benchmark in extracellular vesicles production, whose applicability is limited to lab-scale operations. In this context dead-end and crossflow MF/UF units are compared, both as concentration and purification steps. Followingly, the isolated vesicles ware analysed and characterized with SEC liquid chromatography and both physical and biochemical methods. Finally, passive cargo loading techniques are applied to the isolated vesicles to test their encapsulation capability with bioactive phytocomponents, as curcumin.