Polyethylene glycol-based isolation of urinary extracellular vesicles, an easily adoptable protocol

• Urine can be collected non-invasively and repeatedly, hence a very useful specimen for biomarker discovery. Urinary EVs (UEVs), derived from urine, offer a stable diagnostic tool, but standardised isolation and analysis approaches are warranted. • To have enough UEVs for any study, large volumes of urine sample are necessary, which limits different isolation methods by cost, yield, and time. • The protocol developed could help researchers by offering a cost-effective and dependable UEV isolation method and may lay the foundation for UEVs adoption in clinical space.