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Produktion af Adeno-Associerede Virus Vektorer i Cell Stakke til prækliniske undersøgelser i store dyremodeller
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
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

Automatic Translation

Her giver vi en detaljeret procedure for storstilet produktion af AAV-vektorer af forskningskvalitet ved hjælp af klæbende HEK 293 celler dyrket i cellestakke og affinitetskromatografirensning. Denne protokol giver konsekvent >1 x 1013 vektorgenomer/mL, hvilket giver vektormængder, der er passende til store dyreforsøg.

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