Asymmetric depth-filtration – a versatile and scalable approach for isolation and purification of extracellular vesicles

Abstract

A novel asymmetric depth filtration (DF) approach for isolation of extracellular vesicles (EVs) from biological fluids is presented, and its performance is compared with established methods. The developed workflow is simple, inexpensive, and relatively fast. Compared with ultracentrifugation and size-exclusion chromatography, the developed method isolates EVs with higher purity and yield. Only standard laboratory equipment is needed for its implementation, which makes it suitable for low-resource locations. The described implementation of the method is suitable for EV isolation from small biological samples in diagnostic and treatment guidance applications. Following the scale-up routes adopted in the biomanufacturing of therapeutics, which routinely rely on DF as one of the product purification steps, the developed method may be scaled up to harvesting therapeutic EVs from large volumes of growth medium.

Current advances in non‐viral gene delivery systems: Liposomes versus extracellular vesicles

“…The team demonstrated that DF outperforms ultracentrifugation and SEC in terms of isolation purity and yield. [63] Therefore, this new technique may be used for both diagnostics and EV research.…”

“…This approach is simple, fast, inexpensive and does not require specialized equipment. This method was first proposed by Chernyshev et al [63] In a single-step procedure, plasma was diluted in 1:50 Phosphate-buffered saline, centrifuged at 4500g for 30 min at 4°C, and the supernatant was collected and filtered through a 0.8 μm filter. The collected sample was then concentrated by centrifugation at 3500g using a 100 kDa MWCO filter to isolate EVs.…”