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Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models
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Chapters
- 00:04Introduction
- 00:46Double Plasmid Transfection of HEK293 Cells in Cell Stacks
- 02:30Harvesting Adeno-Associated Virus AAV and Chemical Lysis of The Transfected HEK293 Cells
- 03:23AAV Vector Purification Using Heparin Affinity Chromatography
- 05:45Results: Serum Human Monoclonal Antibody (hIgG) Expression in Animals Upon Administration of AAV6.2FF-hIgG
- 06:41Conclusion
Here we provide a detailed procedure for large-scale production of research-grade AAV vectors using adherent HEK 293 cells grown in cell stacks and affinity chromatography purification. This protocol consistently yields >1 x 1013 vector genomes/mL, providing vector quantities appropriate for large animal studies.
