What they are

Often called plant-derived extracellular vesicles (PDEVs) or plant nanovesicles (PDNVs), these are 30–200+ nm lipid bilayer vesicles released by plants (fruits, leaves, seeds, roots). They carry miRNA/siRNA, lipids, proteins, and metabolites and can cross kingdoms—being taken up by mammalian cells, microbes, and other plants.

Source & Pre-Processing

Sources: Ginger, grape, citrus, broccoli, ginseng, aloe, wheat, rice, tea leaves, etc.

Harvest: Fresh tissues → rinse (low-endotoxin water), homogenize or juice (keep 4 °C), add antioxidant (e.g., 1–5 mM EDTA + 1 mM DTT optional) to curb phenolics.

Clarify: 300–500 g (cell debris) → 2,000–10,000 g (organelles), with protease/PVPP to bind polyphenols; filter 0.8/0.45 µm.

Isolation Methods (choose by scale/purity)

Differential ultracentrifugation (UC)

100,000–150,000 g pellets. Pros: standard; Cons: co-pelleting contaminants.

Density gradients (sucrose or iodixanol)

Best purity; float at 1.08–1.20 g/mL.

Tangential flow filtration (TFF) + size-exclusion chromatography (SEC)

GMP-friendly scaling; SEC gives tight size fractions.

Ultrafiltration (UF)

Fast pre-concentration; pair with SEC/gradient for purity.

Polymer precipitation (PEG)

High yield, lower purity; good for screening.

Aqueous two-phase systems (ATPS)

Gentle; separates from soluble proteins and phenolics.

Immunoaffinity / ligand capture (developing)

Targets plant EV markers (e.g., TET8 in Arabidopsis); currently species-limited.

Microfluidic EV chips

Rapid analytics and small-volume isolation.

Recommended scalable pipeline: Clarification → TFF (300 kDa) → SEC (qEV/packed Sepharose) → optional density polish.