Application of PATfix SEC and CIM Anion Exchange Chromatography for Optimization of EV Upstream Production

Extracellular vesicles (EVs) are a diverse group of particles secreted by all living cells. Numerous different therapeutic applications of both native and engineered EVs are currently in different stages of clinical development. Nevertheless, considerable challenges are still present in the manufacturing, purification and analytics of EVs. Many factors can influence the final product, therefore an all-inclusive approach to development of the bioprocess is needed. Cell culture parameters and production platform selection might alter the number and composition of EVs. Furthermore, raw materials used in upstream production, such as media and supplements, can greatly impact the chromatographic purification.

In this study, we evaluated EV production in different HEK293-derived cell lines. Separation on a strong anion exchange column CIMmultus EV was used to assess the abundance of different EV populations. Multi-detector PATfix SEC analytics coupled with antibody labeling was then used to analyze chromatographic fractions. Furthermore, the analytical methods and performance in downstream processing were applied in the optimization of the upstream process.

 

*Please note that the CIMmultus EV product line referenced in this document has been discontinued as of 1st June 2024. For comparable results, we recommend using products from the CIMmultus QA line.

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