自动化等离子体净化系统可以增加AAV生产
Hannes Thorell1, Sanne Rönning1, Olivér Daoda2
1KTH Royal Institute of Technology, Dept. of Protein science; Stockholm, SE-106 91, Sweden.
New biotechnology
|March 5, 2026
概括
自动化等离子体DNA净化显著减少了腺相关病毒 (AAV) 载体的制造时间. 虽然产量可能会下降,但自动化方法在特定系统中惊人地提高了AAV生产效率.
科学领域:
- 生物技术是生物技术.
- 基因治疗制造业 基因治疗制造业
- 分子生物学分子生物学
背景情况:
- 基因相关病毒 (AAV) 载体对于基因疗法至关重要,但高制造成本阻碍了开发和获取.
- 等离子体DNA (pDNA) 净化是AAV生产中的一个重要瓶,特别是在更大的规模上.
- 目前的手动等离子体净化方法耗时且限制了可扩展性.
研究的目的:
- 为了比较人工与自动化等离子体DNA净化系统在AAV生产中的效率和结果.
- 评估尺度 (Maxi,Mega,Giga) 对净化时间,产量和内毒素水平的影响.
- 调查净化方法之间观察到的AAV生产量差异的潜在机制.
主要方法:
- 使用常规手动套件和可扩展的自动化系统在Maxi,Mega和Giga尺度上的等离子体DNA净化.
- 通过使用纯化的pDNA对HEK293F细胞进行基于PEI的转染来产生AAV载体.
- 使用动态光散射分析对AAV9标位,内毒素水平和pDNA复合体形成进行分析.
主要成果:
- 自动化净化显著减少了处理时间,在更大的规模上节省了更多 (例如,在千兆级节省了8小时).
- 与手工方法相比,自动化系统的结果通常是较低的产量 (减少3-67%) 和更高的内毒素水平.
- 令人惊的是,自动化系统净化的等离子体导致AAV-MAX生产量增加了14倍,这是由于自动化系统的化缓冲器形成的较大的pDNA转染复合体.
结论:
- 自动化等离子体净化为AAV制造提供了相当大的时间节省,特别是在规模上.
- 净化方法的选择及其相关的化缓冲器可以严重影响转化效率和随后的生物制剂生产.
- 优化等离子体制备方法,包括化缓冲成分,有可能改善AAV和其他生物制剂的短暂生产.
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