Chromatographic Tools for Isolation of Extracellular Vesicles and Process Monitoring Analytics

Katja Vrabec, Project Manager, Sartorius BIA Separations

The development of fast and efficient processes for the isolation of extracellular vesicles (EVs) depends on the availability of chromatography media that meet the special fractionation needs of these products and analytical methods for process monitoring. Convective Interaction Media (CIM) monolithic columns offer high binding capacities and low shear stress conditions for the purification of large biologics such as EVs. Multiple-detector PATfix technology was developed for monitoring sample composition and product detection in upstream and downstream processes. We will present the EV isolation process from clarified conditioned media and fractionation using CIMmultus chromatography columns. Testing of the raw materials, analysis of upstream samples produced in different cell lines, and analysis of downstream samples were performed using PATfix analytical approaches. The composition of EV populations was monitored through the detection of tetraspanins using immunofluorescent labelling and SEC analytics. High-throughput analysis of in-process samples and impurity composition was monitored using CIMac anion exchange columns.

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