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Chapters
- 00:05Introduction
- 00:44Generation of Baculovirus-infected TIPS Cells
- 03:05Production of AAV Vector
- 05:08Lysis of Cells and Release of AAV
- 06:07Purification of AAV Vector Using Affinity Column Chromatography System
- 08:03Concentration and Diafiltration of AAV Vector Using Tangential Flow Filtration (TFF)
- 08:54Results: Purification of Adeno-Associated Virus (AAV)2 Vector Using Baculovirus-Insect Cell Culture System
- 09:48Conclusion
I detta protokoll produceras AAV2 vektor genom samodling Spodoptera frugiperda (Sf9) insektsceller med baculovirus (BV)-AAV2-grönt fluorescerande protein (GFP) eller terapeutisk gen och BV-AAV2-rep-cap infekterade Sf9 celler i suspension kultur. AAV-partiklar frigörs från cellerna med tvättmedel, förtydligas, renas genom affinitetskolonnkromatografi och koncentreras genom tangentiell flödesfiltrering.
