Adeno-Associated Viruses (AAVs) are increasingly receiving approval for the in vivo treatment of acquired and inherited genetic diseases. However, purification challenges remain as AAV production typically results in a significant proportion of empty capsids which may invoke an immune response, without the ability to deliver the gene therapy payload.<br/>Here we propose a scalable purification process, based on anion exchange (AEX) membrane chromatography for final polishing and empty/full capsid separation. Despite widespread application of AEX membrane chromatography, there has been little published effort to understand the scalability of membrane devices when transitioning from filter plate experiments to flow capsules. A Design of Experiments (DoE) approach was implemented for the optimization of binding and elution conditions of AAVs via high throughput screening on Mustang Q filter plates. These conditions were transferred to Mustang Q capsules along with alternative membrane chromatography devices of comparable scale. Gradient and step elution experiments were carried out to understand the role of salt and additives in desorption of AAV from the membrane surface and the separation of empty and full capsids. In summary, a scalable and platform-ready AAV purification protocol was developed to facilitate the transition from Mustang Q filter plates to XT acrodiscs.

Scalability and competitor benchmarking of anion exchange membrane chromatography devices for adeno-associated virus purification using a DoE approach

ACS Fall 2020 Virtual Meeting

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.