Alternative solutions to separate AAV full and empty capsids using anion exchange

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.

In this webinar, a high-performance anion exchange (AEX) chromatography separation for several AAV serotypes using step-elution mode is presented, and 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. The reproducibility and scale up potential of these AEX approaches will also be demonstrated.

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.

Attend this webinar to:

• 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.
• Discover hints and tips to avoid common pitfalls

Åsa Hagner-McWhirter

Principal Scientist at Cytiva

Åsa has been with Cytiva based in Uppsala, Sweden since 2003 and is a Downstream and analytics SME, with a broad and deep understanding of viral vector processing. Due to her long experience and from customer interactions she has gained insights into common challenges and pitfalls in the area of viral vectors and vaccines as well as general protein purification and analysis. She has also worked with proteomics and fluorescent-based protein analysis technologies.

Åsa holds a PhD in Medical Biochemistry from Uppsala University in 1999 based on research around biosynthesis of proteoglycans. The studies involved polysaccharide structure analysis, enzyme purification, and cloning as well as characterizing an enzyme reaction.

Mark Schofield

Senior R&D Manager at Cytiva

Mark earned his degrees in Scotland, a bachelor’s degree from the University of Edinburgh and a molecular biology PhD from the University of Dundee. For the last 12 years he has been at Cytiva focusing on chromatography applications. Currently his team works on bioprocess intensification solutions and chromatographic separations for gene therapy modalities.

Watch now

SPEAKERS

Åsa Hagner-McWhirter

Principal Scientist at Cytiva

Mark Schofield

Senior R&D Manager at Cytiva

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