Technology and Process Development (TPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China.
Protein Expr Purif. 2021 Dec;188:105976. doi: 10.1016/j.pep.2021.105976. Epub 2021 Sep 16.
In general, purification of bispecific antibody (bsAb) is more challenging than that of monospecific antibody due to the increased complexity in byproduct profile. Like in the case of monospecific antibody purification, immunoglobulin-binding protein-based affinity chromatography is an indispensable tool for bsAb purification. For example, Protein A affinity chromatography has been widely used to capture Fc-containing bsAbs whereas other affinity media such as Protein L and KappaSelect, which bind kappa light chain, are used to capture bsAbs that do not contain a Protein A-binding site. In fact, affinity chromatography also possesses the capability of removing certain product-related impurities in bsAb purification when it is conducted with suitable medium and under appropriate conditions. Fully exploring the potential of affinity chromatography in bsAb purification to achieve both product capture and byproduct removal is highly desirable, as this can greatly alleviate the purification burden on subsequent polishing steps and hence improves the overall robustness of the downstream process. This article briefly reviews the byproduct clearance potential of several commonly used affinity media under relevant bsAb purification scenarios.
一般来说，由于副产物谱的复杂性增加，双特异性抗体（bsAb）的纯化比单特异性抗体更具挑战性。与单特异性抗体的纯化一样，基于免疫球蛋白结合蛋白的亲和层析是 bsAb 纯化不可或缺的工具。例如，Protein A 亲和层析已广泛用于捕获含 Fc 的 bsAb，而其他亲和介质，如结合 κ 轻链的 Protein L 和 KappaSelect，则用于捕获不含 Protein A 结合位点的 bsAb。事实上，当在合适的介质和条件下进行亲和层析时，它还具有去除 bsAb 纯化中某些与产品相关的杂质的能力。充分挖掘亲和层析在 bsAb 纯化中的潜力，实现产物捕获和副产物去除，是非常理想的，因为这可以大大减轻后续精修步骤的纯化负担，从而提高下游工艺的整体稳健性。本文简要综述了几种常用亲和介质在相关 bsAb 纯化方案下的副产物清除潜力。
