Zhang Yuan, Wang Ying, Li Yifeng
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.
Protein Expr Purif. 2019 Jun;158:65-73. doi: 10.1016/j.pep.2019.02.017. Epub 2019 Mar 1.
Protein A chromatography is generally less effective at removing antibody aggregates under typical conditions. We recently developed a method that can significantly improve Protein A's aggregate removal capability. This method involves adding calcium chloride/polyethylene glycol (PEG) or sodium chloride/PEG combination to wash and elution buffers. Each salt alone showed some resolution-enhancing effect when its concentration reaches a certain level, and this effect was significantly enhanced by the presence of PEG, which itself had no effect on resolution. The synergistic effect of salt and PEG results in almost complete separation of monomer from aggregates. For the particular case used for method development and demonstration, in comparison with the control run the optimized procedure reduces aggregates in elution pool from 20% to 3-4%. This new method, by facilitating aggregate removal at the capture step, improves the overall robustness of downstream process.
在典型条件下，蛋白A层析在去除抗体聚集体方面通常效果较差。我们最近开发了一种方法，可显著提高蛋白A去除聚集体的能力。该方法包括在洗涤缓冲液和洗脱缓冲液中添加氯化钙/聚乙二醇（PEG）或氯化钠/PEG组合。当每种盐的浓度达到一定水平时，单独使用时都显示出一定的分辨率增强效果，并且PEG的存在显著增强了这种效果，而PEG本身对分辨率没有影响。盐和PEG的协同作用几乎能使单体与聚集体完全分离。对于用于方法开发和验证的特定案例，与对照运行相比，优化后的程序将洗脱液中的聚集体从20%减少到3 - 4%。这种新方法通过在捕获步骤促进聚集体的去除，提高了下游工艺的整体稳健性。
