过程优化减轻了Fc融合蛋白在超过/透过过程中的保留损失
Biotechnology progress
|March 12, 2025
概括
超过/透过 (UF/DF) 对于抗体治疗至关重要. 过程优化,考虑到pH和度两极分化,改善了Fc-fusion蛋白质保留超出了膜分子重量切割值,提高了产量和质量.
科学领域:
- 生物制药制造业 生物制药制造业
- 蛋白质化学 蛋白质化学
- 分离科学 分离科学
背景情况:
- 超过/透过 (UF/DF) 是基于抗体的治疗药物的标准下游过程,对于度和缓冲区交换至关重要.
- 膜选择,料流量和跨膜压力 (TMP) 是影响UF/DF性能的关键因素,影响产量,缓冲区交换和产品质量.
研究的目的:
- 调查超出分子重量截止值 (MWCO) 的因素,这些因素会影响 UF/DF 期间的 Fc 融合蛋白保留.
- 优化 UF/DF 过程参数,以提高 Fc 融合蛋白质的回收和质量.
主要方法:
- 使用尺寸排除色谱 (SEC) 来评估Fc融合蛋白的构成和保留.
- 优化了UF/DF参数,包括膜类型,pH,料流量和跨膜压力 (TMP).
主要成果:
- 由于pH诱导的形状变化,Fc融合蛋白的保留受到载荷材料的pH值的显著影响.
- 通过增加料流量和降低TMP来减少度极化,使透物中蛋白质损失最小化.
- 使用 5 kDa 或 10 kDa 膜在优化过程条件下实现了高保留率.
结论:
- 在UF/DF过程中,Fc融合蛋白的保留受到MWCO以外的因素的影响,特别是pH依赖的形状变化和度极化.
- 优化过程参数,如负载条件,料流量和TMP,对于最大限度地恢复蛋白质和确保UF/DF中的产品质量至关重要.
- 这些发现为改进生物制药下游加工的UF/DF策略提供了宝贵的见解.
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