Adeno-associated virus (AAV) is the main vector for gene therapy and there is need for scalable, cost-efficient, and robust chromatography-based purification. Key for a successful process are high overall yields of full capsids with effective empty capsid reduction and efficient impurity removal.
Here we present high-performance anion exchange (AEX) chromatography separation for several AAV serotypes using step-elution mode. Optimized protocols and conditions for both resin and membrane formats are described. Full and empty capsid peaks were identified using the UV260:280 ratio from chromatograms. And separation results were confirmed by PCR, ELISA, mass photometry, and analytical ultracentrifugation. We show that these AEX approaches are reproducible and can be scaled up.
In conclusion, to maximize full and empty capsid separation by AEX, conditions such as pH, MgCl2, elution salt concentration, and length of the elution step are critical. Depending on serotype or AAV capsid properties, prescreening and fine-tuning for the optimal conditions may be required, but high yields and significant enrichment of full capsids can be achieved.
In this webinar you'll:
• Learn about AAV full and empty capsid separation using Capto Q and Mustang Q anion exchange chromatography.
• Dive into details on how to achieve high yields and purity of full capsid.
• Understand the pre-screening procedure that is needed for each AAV capsid serotype/variant.
Have more questions? Let us know how we can help.
https://info.cytivalifesciences.com/vv-downstream-request-contact.html
