维护rAAV稳定的关键方面
Johannes Lengler1, Mia Gavrila1, Janina Brandis2
1Gene Therapy Process Development, Baxalta Innovations GmbH, a Part of Takeda Companies, Uferstraße 15, 2304, Orth an Der Donau, Austria.
Scientific reports
|November 12, 2024
概括
保持腺相关病毒 (AAV) 载体的稳定性对于基因治疗至关重要. 储存AAV载体在-65°C以下或使用冷化,可以长时间保持它们的生物潜能和疗效.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 基因治疗 基因治疗
背景情况:
- 腺相关病毒 (AAV) 载体对于基因疗法应用至关重要.
- 在储存期间确保AAV载体的产品质量和有效性是一致的,这一点至关重要.
- 了解存储条件对AAV载体特性的影响,对于制造和临床使用至关重要.
研究的目的:
- 研究各种储存条件对rAAV8和rAAV9载体物理化学和生物特性的影响.
- 评估AAV制剂的短期存储稳定性,低温生存性和冷解效应.
- 确定最佳的储存策略,以保持AAV载体的生物潜能和疗效.
主要方法:
- 体和基因组标位的量化.
- 生物潜能评估,以评估载体的功能性和有效性.
- 在不同的温度,pH值和储存条件下 (短期,长期,冷/解,冷化) 分析AAV载体稳定性.
主要成果:
- 冷/解周期对rAAV9-aGAL的功能影响最小.
- 长时间的室温储存降低了生物潜能,而高温加速了衰变.
- 添加糖和酸醇增强了rAAV稳定性;酸性pH (2.5) 显著降低了生物潜能.
- 在 -65°C以下的长期储存保存了rAAV载体的有效性多年;冷解保持了至少10个月的功能.
结论:
- 储存温度和pH值显著影响AAV载体的生物潜能和疗效.
- 非常低温储存 (低于-65°C) 和冷化是长期保存AAV载体的有效方法.
- 单靠囊酸标位并不能保证生物效能保持;功能性评估至关重要.
- 优化的储存条件可以最大限度地提高AAV载体的稳定性和治疗潜力.
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