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使用杆状病毒-昆虫细胞培养系统生产和纯化腺相关病毒(AAV)2载体的工艺开发
JoVE Journal
Biology
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JoVE Journal Biology
Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System
DOI:

10:31 min

January 13, 2022

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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

Summary

Automatic Translation

在该协议中,通过将 Spodoptera frugiperda(Sf9) 昆虫细胞与杆状病毒(BV)-AAV2-绿色荧光蛋白(GFP)或治疗基因和BV-AAV2-rep-cap感染的Sf9细胞在悬浮培养物中共同培养来产生AAV2载体。使用洗涤剂从细胞中释放AAV颗粒,澄清,通过亲和柱色谱纯化,并通过切向流过滤浓缩。

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