Development and Optimization of a Serotype-Independent Method of Adeno-Associated Virus (AAV) Harvest and Purification (SJ-16-0036)

St. Jude Reference #SJ-16-0036

Description

AAV vectors are used in over 7% of all gene therapy clinical trials. During production, AAV is either released into the cell culture media or retained within the cell. Large-scale physical lysis methods for the release of cell-associated AAV require a significant capital investment and are not easily utilized when producer cells are grown within a solid support such as a perfusion bioreactor. Chemical methods, such as using detergent to release AAV from cells, can be highly successful. However, the detergent then becomes an impurity that has to be removed to acceptable limits and poses a challenge to downstream processing.

Researchers at St. Jude have developed a novel, integrated method for releasing cell associated AAV from producer cells and enriching this released AAV. The addition of ammonium sulfate to cells leads to osmotic shock and cellular lysis, releasing cell-associated AAV. Simultaneously, the ammonium sulfate solution acts as a chaotropic agent to promote binding between AAV and hydrophobic interaction chromatography ligands. The phenyl ligand provides highly effective binding and recovery of multiple serotypes of AAV from both cell culture media and cell lysates. This process leads to a substantial reduction of dsDNA-containing insoluble impurities, as well as host cell proteins. This has important implications for large scale production as this technique is amenable to either suspension or fixed bed adherent cell culture. This simple, robust, and scalable process integrates release, capture, and enrichment of AAV vectors and is compatible with multiple industrially relevant serotypes of AAV.


 

Keywords

AAV, Gene therapy, vector production


 

Granted Patents or Published Applications

Pending


 

Related Scientific References


 

Licensing Opportunities

The invention can be used for harvest and downstream processing of clinical-grade AAV gene therapy vectors, and to prepare AAV for use in research and development settings. It was developed for the production and purification of an AAV Factor IX vector, and has been nonexclusively licensed in that field; however, it can be used and is available for license in the production of any AAV vector. Contact: chad.riggs@stjude.org.

Related Links

Contact the Office of Technology Licensing (Phone: 901-595-2342, Fax: 901-595-3148) for more information.