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Procesudvikling til produktion og rensning af Adeno-Associated Virus (AAV)2 Vector ved hjælp af Baculovirus-Insect Cell Culture System
Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP)
Journal (Biology)
Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP)
Engineering Cell-permeable Protein
Journal (Biology)
Engineering Cell-permeable Protein
 
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Procesudvikling til produktion og rensning af Adeno-Associated Virus (AAV)2 Vector ved hjælp af Baculovirus-Insect Cell Culture System

Article DOI: 10.3791/62829-v • 10:31 min • January 13th, 2022
January 13th, 2022 •
1,2, 1, 1,2,3, 1
1Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, 2University of Cincinnati College of Medicine, 3Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children's Hospital of Philadelphia

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I denne protokol produceres AAV2 vektor af co-culturing Spodoptera frugiperda (Sf9) insektceller med baculovirus (BV)-AAV2-grønt fluorescerende protein (GFP) eller terapeutisk gen og BV-AAV2-rep-cap inficerede Sf9-celler i suspension kultur. AAV partikler frigives fra cellerne ved hjælp af vaskemiddel, afklaret, renset ved affinitet kolonne kromatografi, og koncentreret ved tangentiel flow filtrering.

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Biologi Udgave 179 AAV baculovirus Sf9 celler bioreaktor kromatografi rensning tangentiel flow filtrering
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