改进了单克隆抗体电荷变异分离的工艺设计,使用多列逆流溶剂梯度净化多列
Shu-Ying Jing1, Ce Shi1, Dong Gao2
1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang Key Laboratory of Smart Biomaterials, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China.
Journal of chromatography. A
|August 16, 2023
概括
一个新的重新设计策略改善了蛋白质的多列逆流溶剂梯度净化 (MCSGP). 这种方法通过创建更准确的化配置文件来提高回收和纯度,克服了传统批量工艺设计的局限性.
科学领域:
- 生物技术是生物技术.
- 化学工程是化学工程的重要组成部分.
- 染色体学 染色体学 是一种染色学.
背景情况:
- 多列逆流溶剂梯度净化 (MCSGP) 是有效的分离蛋白质与重叠的化配置文件.
- 由于流量和蛋白质负载的变化,批处理设计与实际MCSGP条件之间的差异会降低目标蛋白质的纯度和回收.
研究的目的:
- 为MCSGP开发一个改进的工艺设计 (重新设计),从而产生更具代表性的化配置文件.
- 通过更好地模拟连续工艺条件来提高MCSGP的分离性能.
主要方法:
- 使用首次运行的MCSGP实现了一个新的重新设计策略,以捕获实际的连续工艺化配置文件.
- 基于重新设计的MCSGP (reMCSGP) 使用模型蛋白质混合物和单克隆抗体 (mAb) 充电变体进行了验证.
主要成果:
- 对于一个模型蛋白质混合物,reMCSGP将回收率从83.6%提高到97.8%,同时保持>95%的纯度.
- 对于mAb充电变体,reMCSGP在>84%的纯度下实现了93.9%的回收,超过了标准的MCSGP.
结论:
- 重新设计策略提供了一个更准确的化配置文件,它密切反映了连续工艺条件.
- 这种增强的工艺设计显著提高了MCSGP的分离性能,纯度和回收.
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