Isolation of extracellular vesicle from blood plasma using electrophoretic migration through porous membrane

Abstract

We present a high-yield electro-migration method to isolate extracellular vesicles (EVs), which achieves ultracentrifugation (U/C)-level size exclusion from biological fluids (e.g.,plasma).An electric field applied across a dialysis membrane with an appropriate pore size (30nm), facilitates protein migration through the membrane, but captures EVs on the membrane surface. Compared to conventional procedures, this method shows up to 65% EV recovery estimated at the RNA level (similar to 7.9 times better than U/C) and up to 83.6% protein removal (residual protein amount is approximately half of the precipitate) in similar to 30 min (similar to 9 times faster than U/C). With optimized working range of voltage and use of biochemically stable buffers, isolated EV are fully compatible with biological post-processes and assays. (C) 2016 Elsevier B.V. All rights reserved.