Ivano Luigi Colao, Randolph L. Corteling, Daniel G. Bracewell, Ivan B. Wall
Cytotherapy 2024
Abstract:
Extracellular vesicles (EVs) have emerged as promising cell-free therapeutic candidates. A key focus of EV research is the development of scalable and robust purification methods. However, there is limited literature evaluating these methods in relation to the potency of the EV product.
This study examines two monolith chromatography methods to assess the ability of purified EVs to retain their stimulatory effects on fibroblasts, thereby linking scalable purification methods with product outcomes. The study found that monolith chromatography, using both QA (Quaternary Amine) and OH (Hydrophobic Interaction) columns, effectively purified EVs from neural stem cell cultures. Tangential flow filtration (TFF) was employed as a pre-purification step, facilitating the recovery of functional EVs. During QA chromatography, EVs co-eluted with albumin, indicating some co-isolation of proteins. The OH column demonstrated superior performance in terms of capacity and purity, capturing more EVs with fewer impurities. Both columns, whether used alone or in combination, maintained the EVs’ functional activity in stimulating fibroblast wound repair in an in vitro scratch assay. This research supports the use of monolith chromatography, combined with TFF, as a scalable purification method that preserves the biological activity of EVs, making it suitable for therapeutic applications.
