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Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models
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JoVE 杂志 免疫与感染
Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models
DOI:

07:21 min

June 30, 2021

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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

Summary

自动翻译

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.

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