多参数过程优化用于高产量AAV2载体生产,使用缩放式多平行生物反应器系统和高通量分析工具
Nazgul Wagner1, Jonas Austerjost1, Julia Niemann1
1Sartorius Stedim Biotech GmbH, August-Spindler-Straße 11, Göttingen 37079, Germany.
Journal of biotechnology
|December 21, 2025
概括
使用生物反应器优化腺相关病毒 (AAV) 生产,揭示了温度变化显著提高功能性AAV产量. 批处理过程的效率增加了两倍,而 perfusion 过程的效率增加了三倍以上,从而提高了整体效率.
科学领域:
- 生物技术是生物技术.
- 基因治疗 基因治疗
- 生物工艺工程 生物工艺工程
背景情况:
- 腺相关病毒 (AAV) 载体对于基因疗法提供至关重要.
- 过渡性生产是AAV制造的常见但可变的方法.
- 优化过程参数对于高效的AAV矢量生产至关重要.
研究的目的:
- 为了评估腺相关病毒 (AAV) 生产的关键过程参数.
- 为了比较AAV产量的批量与 perfusion 生物工艺模式.
- 调查温度变化对AAV生产效率的影响.
主要方法:
- 利用一个15毫升的多平行缩放式生物反应器系统来评估细胞密度和等离子体DNA度.
- 采用250毫升生物反应器来比较批量和 perfusion 过程.
- 研究了温度转移策略以提高AAV产量.
- 评估生产使用生物层干涉计 (BLI) 进行囊位和Incucyte®用于功能位.
主要成果:
- 在低细胞密度条件下,在37°C的批处理过程中, perfusion 过程的功能位数比批处理过程低三倍以上.
- 温度变化使功能性AAV产量在批量中增加了近两倍,在 perfusion 过程中增加了三倍以上.
- 温度变化改善了功能和体标位的比率,表明化增强.
结论:
- 优化的生物反应器条件,包括温度变化,显著提高功能性AAV产量.
- 这项研究表明了优化的AAV生产过程的可扩展性和可重复性.
- 高通量分析技术可以快速评估AAV生产质量.
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