实施多式离子交换染色学以应对产品质量挑战和下游平台的局限性:一个案例研究
Julie Robinson1, Mayank Vats1, Michael Hartmann2
1Biologics Process Research and Development, Merck & Co., Inc., Rahway NJ 07065 USA.
Journal of chromatography. A
|February 21, 2025
概括
多模态离子交换染色学 (MMAEX) 为净化复杂的Fc融合蛋白提供了强大的解决方案,克服了传统方法的局限性. 这项研究证明了MMAEXEX.
科学领域:
- 生物制药工艺开发 生物制药过程开发
- 染色学净化 染色学净化
- 蛋白质工程是指蛋白质工程.
背景情况:
- 传统的离子交换染色学 (AEX) 在去除复杂生物分子中的杂质方面面临挑战,例如低PI Fc融合蛋白.
- 病毒清除要求可以限制传统AEX抛光步骤的设计空间.
- 多模态离子交换染色学 (MMAEX) 为类似平台的制造工艺提供了一个有前途的替代方案.
研究的目的:
- 使用MMAEX.开发一个优化的抛光步骤,用于具有挑战性杂质配置的Fc融合蛋白,使用MMAEX.
- 为了评估MMAEX Capto Adhere的性能,作为传统AEX流通抛光的替代品.
- 调查工艺参数和料成分对MMAEX步骤性能的影响.
主要方法:
- 综合计算和高通量选 (HTS) 工作流程用于树脂和条件选.
- 在pH值5.5.5时对MMAEX Capto Adhere进行实验评估.
- 基于产量,总清除,宿主细胞蛋白 (HCP),DNA和病毒清除 (XMULV,MVM) 的阶段性能的评估.
主要成果:
- 在pH 5.5的MMAEX Capto Adhere被确定为Fc融合蛋白的可行抛光步骤.
- HMW清除受初始总量水平的影响,在中等负载时清除最佳.
- 其余的HCP和DNA被有效地去除到量化限制以下.
- 证明了XMULV和MVM的4个日志的病毒清除,以及与过程相关的料流.
结论:
- 集成在模拟和HTS简化生物工艺开发和抛光步骤优化.
- MMAEX提供了一种灵活有效的平台,用于净化复杂的Fc融合蛋白.
- 料杂质的成分显著影响MMAEX性能,需要仔细的工艺设计.
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