For recombinant antibody purification, removal of product-related impurities usually relies on the two polishing steps post Protein A chromatography. A certain impurity may bind weaker or tighter to a particular type of column than the target antibody, and this forms the basis for separation. For impurities that bind weaker, they can be removed by pre-elution wash under appropriate conditions. For impurities that bind stronger, they can be separated by using a suitable condition that selectively elutes the product. In this study, with a bispecific antibody case, we compared the relative robustness of byproduct removal by wash and by elution using two different types of chromatography. The data suggest that elution-enabled byproduct clearance is more robust than wash-enabled clearance, and the former approach provides consistent impurity clearance over a relatively wide range of loading density.

对于重组抗体纯化，产品相关杂质的去除通常取决于蛋白A层析后的两个抛光步骤。某些杂质可能会比目标抗体更弱或更紧密地结合到特定类型的色谱柱上，这构成了分离的基础。对于结合力较弱的杂质，可以在适当的条件下通过预洗脱洗涤将其除去。对于结合力更强的杂质，可以使用选择性洗脱产物的合适条件将其分离。在这项研究中，以双特异性抗体为例，我们比较了使用两种不同类型的色谱法通过洗涤和洗脱去除副产物的相对耐用性。数据表明，可洗脱的副产物清除能力比可洗涤的清除能力强，