April 5, 2024

Exosome isolation by tangential flow filtration and size exclusion chromatography

We demonstrate a scalable workflow for the isolation of exosomes—a type of extracellular vesicle with significant potential for targeted drug/gene delivery. For research and development and diagnostic purposes, ultracentrifugation is an established technology to separate and purify exosomes. However, ultracentrifugation is not easily scalable. We describe an evaluation of purification workflows from three different cell culture supernatants. The workflows we describe are suitable for scale-up and combine tangential flow filtration (TFF) for the concentration of exosomes followed by gentle size exclusion chromatography (SEC) with our next-generation Cytiva™ superSEC resin. We found that:

- Cytiva™ superSEC resin enriched exosomes from impurities compared to legacy Sepharose™ CL-2B resin—a well-established resin for exosome purification—after the initial TFF step.

- High productivity, scalability, and single-use compatibility were observed for the superSEC resin.

- Scaling up the SEC resin volumes using the superSEC resin was clearly shown with shorter purification cycle times than used in existing SEC protocols for the purification of exosomes.

Acknowledgements to VivaZome Therapeutics Pty Ltd and Evox Therapeutics Ltd for contributions to this application note.


Tags: downstream, gene therapy, process development, purification, size exclusion chromatography, Exosome, TFF