Free flow electrophoresis allows quick and reproducible preparation of extracellular vesicles from conditioned cell culture media

Aim: Despite intensive research during the last decade, it remains challenging to prepare extracellular vesicles (EVs) of high purity, especially from primary body liquids or protein-rich conditioned media. For now, time-consuming combinations at least two orthogonal methods, e.g., density and size separation, are required enrich EVs often expense processing time. Therefore, novel technologies that allow EV preparation in acceptable time intervals fair purities. Free-flow electrophoresis (FFE) constitutes a well-established semi-preparative method separate analytes, by inherent differences their electric charges. FFE combines flow-driven longitudinal transport sample material with vertical allows separation components into up 96 different fractions. It was our aim evaluate potential for other EV-containing cell culture Methods: Exemplarily, media mesenchymal stem/stromal cells raised presence 10% human platelet lysate were processed. We analyzed obtained fractions technologies, including imaging flow cytometry, western blot nanoparticle tracking analysis. Results: demonstrate quickly reproducibly separates huge proportion molecules included original sample. Conclusion: Our results qualify as feasible, quick reproducible technology bona fide EVs.