(340b) Isolation Strategies Based on Membrane Processes for Extracellular Vesicles from Citrus Limon

Extracellular vesicles (EVs) are lipidic nanoparticles secreted by all kind of cells. EVs can transport a pool of different biomolecules between close and distal cells, in a site-specific way. They are gaining increasing attention for their potential use in nanomedicine, nutraceuticals and cosmetics, in particular as drug delivery vectors, diagnostic tools and in tissue regeneration. The discovery that also plants do release EVs opened the doors to the use of vegetal vesicles as carriers of bioactive compounds and genes.

The greatest challenge in the field is associated to EVs separation from complex biological fluids. Ultracentrifugation is considered as the golden standard but it is mostly a lab-scale technique with a very low scalability potential. Conversely, alternative techniques as liquid chromatography and filtration processes are rising, allowing to obtain high yield products and feasible application to industrial scale.

This work is aimed at the development of a scalable and inexpensive integrated membrane process for the isolation of EVs from plants and fruits. Citrus Limon is selected as source material, due to its beneficial properties on human health, besides being a cheap and highly abundant source. Indeed, Citrus Limon juice extracts, represent an innovative and still poorly studied source of EVs that have very promising therapeutic use as they inhibit cancer cell proliferation on different cell lines.

In this process both dead-end and crossflow MF/UF units are compared, both as concentration and purification steps. Followingly, the isolated vesicles are analyzed and characterized with SEC liquid chromatography and both physical and biochemical methods. Finally, active and passive cargo loading techniques are applied to the isolated vesicles to test their encapsulation capability with bioactive phytocomponents, as curcumin.