提高带有阴离子缓冲器的阴离子交换色谱的工艺稳定性,以减少聚合物
Shuo Wang1, Shuaihua Wang1, Dijing Shi1
1Downstream Process Development (DSPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.
Protein expression and purification
|December 29, 2024
概括
用于重组蛋白净化的阴离子交换染色学 (CEX) 使用像histidine或Bis-Tris.这样的阴离子缓冲器更强大. 这些缓冲器确保一致的产量和化量,与化不同,改善CEX过程的稳定性.
科学领域:
- 生物化学 生物化学
- 化学工程是化学工程的重要组成部分.
- 生物制药制造业 生物制药制造业
背景情况:
- 阴阳交换色谱 (CEX) 对于在重组蛋白质生产中将蛋白质聚合物与单体分离至关重要.
- 聚合物和单体的相似的单电点挑战了逐步化CEX方法的稳定性.
研究的目的:
- 为了比较化阶段化对化 (histidine和Bis-Tris) 的性能稳定性与CEX中的分阶段化对比.
- 确定CEX化策略,以提高复合蛋白制造过程的稳定性和一致性.
主要方法:
- 利用蒙特卡洛模拟来建模和比较化策略.
- 通过在不同条件下评估单体纯度,产量和化量来评估过程稳定性.
- 研究了pH值和化剂度变化对CEX性能的影响.
主要成果:
- 所有测试的化方法都实现了可接受的单体纯度.
- 化溶解显示出由于pH值和度的变化,产量和溶解体积的显著波动.
- 希斯蒂丁和Bis-Tris缓冲化在广泛的操作范围内表现出一致的产量和化量.
结论:
- 与化相比,阴离子缓冲器 (histidine,Bis-Tris) 为重组蛋白净化提供了更强大的CEX过程.
- 阴离子缓冲器的解离行为有效地减少了pH值变化对离子交换平衡的影响.
- 实施阳离子缓冲化提高了工业规模CEX净化稳定性和可靠性.
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