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蛋白A洗涤缓冲液中用于有效清除宿主细胞蛋白的适量氯化钠。

The adequate amount of sodium chloride in Protein A wash buffer for effective host cell protein clearance.

作者信息

Han Jinyu, Yang Ju, Wang Ying, Li Yifeng

机构信息

Technology and Process Development (TPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.

Technology and Process Development (TPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.

出版信息

Protein Expr Purif. 2019 Jun;158:59-64. doi: 10.1016/j.pep.2019.02.016. Epub 2019 Feb 28.

Abstract

Post-load column wash in Protein A chromatography can effectively improve host cell protein (HCP) clearance. A commonly used wash additive for this purpose is sodium chloride. However, the adequate amount of sodium chloride required for effective HCP clearance is less consistent in literature. In this study we investigated the impact of different amounts of sodium chloride on HCP clearance with five monoclonal antibodies (mAbs). For each mAb, elution pool HCP levels from runs under different wash conditions are compared. For all five mAbs, the data suggested that 250 mM would be an adequate amount for the salt to largely achieve its HCP reducing effect. The same conclusion is also reached for calcium chloride, a less commonly used but equally effective Protein A wash additive for HCP clearance.

摘要

蛋白A层析中的上样后柱洗涤可有效提高宿主细胞蛋白(HCP)清除率。为此常用的洗涤添加剂是氯化钠。然而,文献中关于有效清除HCP所需的适量氯化钠并不一致。在本研究中,我们用五种单克隆抗体(mAb)研究了不同量的氯化钠对HCP清除率的影响。对于每种mAb,比较了不同洗涤条件下运行的洗脱池HCP水平。对于所有五种mAb,数据表明250 mM的盐量足以在很大程度上实现其降低HCP的效果。对于氯化钙也得出了相同的结论,氯化钙是一种较少使用但同样有效的用于清除HCP的蛋白A洗涤添加剂。

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