Production and Purification of Mesenchymal Stem Cell-Derived Extracellular Vesicles Using Preferential Exclusion Chromatography

Abstract

Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) therapeutics hold the potential to harness the regenerative power of their parental cells, promoting proliferation and reducing inflammation. There is a need for the production and isolation process that can be easily scaled and deliver the desired quality and quantity of EVs. We present a chromatographic method development based on preferential exclusion principle that can be used for EV isolation from adherent and microcarrier-based upstream. The method employs monolithic hydroxyl columns to purify and concentrate MSC-EVs. The heterogeneity in EV size and composition poses challenges in upstream and downstream processes; therefore, process development was guided by PATfix biochromatography analytics, tracking EV-specific surface antigens. The developed purification method enables direct concentration of MSC-EVs from conditioned media while removing 97% of protein impurities and 95% of double-stranded deoxyribonucleic acid (dsDNA)-related impurities in the process.