Dong Wanyuan, Zhang Penglong, Wu Di, Wan Yan, Li Yifeng
Downstream Process Development (DSPD), WuXi Biologics, Shanghai, 200131, China.
J Biol Methods. 2024 Dec 12;12(1):e99010045. doi: 10.14440/jbm.2025.0106. eCollection 2025.
Protein L-conjugated resins are affinity media that bind to the variable region of the kappa light chain (LC) and have been used for initial product capture in the downstream processing of full-length antibodies and antibody fragments. Previous studies, including ours, have demonstrated that Protein L chromatography effectively separated various byproducts generated during the production of bispecific antibodies (bsAbs), including half-antibody, homodimer, LC-missing species, and aggregates. Cytiva recently launched its second-generation Protein L resin, MabSelect VL, which offers significantly improved binding capacity compared to its predecessor, Capto L.
This study aimed to explore the feasibility of developing a two-column process, which includes MabSelect VL capture step and a polishing step, for purification of complex antibody molecules.
We employed two bsAb cases to demonstrate that MabSelect VL's enhanced byproduct removal capability allows for a potential two-column purification process.
For both bsAbs, the developed two-column process yielded a product with quality attributes comparable to those obtained using the traditional three-column process.
The MabSelect VL-based two-column process can be successfully applied to bsAb purification. In addition, it should also be feasible with regular monoclonal antibodies, whose purification is generally less challenging than that of bsAbs. By reducing the downstream process from three columns to two columns, significant savings in terms of time, labor, and materials can be achieved.
蛋白L偶联树脂是一种亲和介质，可与κ轻链（LC）的可变区结合，已用于全长抗体和抗体片段下游加工中的初始产物捕获。包括我们的研究在内的先前研究表明，蛋白L色谱法可有效分离双特异性抗体（bsAb）生产过程中产生的各种副产物，包括半抗体、同二聚体、缺失轻链的物种和聚集体。赛多利斯最近推出了其第二代蛋白L树脂MabSelect VL，与前代产品Capto L相比，其结合能力有显著提高。
本研究旨在探索开发一种两柱法（包括MabSelect VL捕获步骤和精制步骤）用于纯化复杂抗体分子的可行性。
我们采用两个双特异性抗体案例来证明MabSelect VL增强的副产物去除能力允许潜在的两柱纯化工艺。
对于这两种双特异性抗体，所开发的两柱法产生的产品质量属性与使用传统三柱法获得的产品相当。
基于MabSelect VL的两柱法可成功应用于双特异性抗体的纯化。此外，对于常规单克隆抗体来说应该也是可行的，其纯化通常比双特异性抗体的纯化难度小。通过将下游工艺从三柱减少到两柱，可以在时间、劳动力和材料方面实现显著节省。
