Chen Xiujuan, Wang Ying, 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. 2020 Nov;175:105711. doi: 10.1016/j.pep.2020.105711. Epub 2020 Jul 29.
Asymmetric IgG-like bispecific antibodies (bsAbs) are normally derived from two parental mAbs with different origin. Despite the implementation of heterodimerization-promoting strategy in the design, homodimerization can still occur at a low level during the recombinant production of these molecules. In general, monitoring and removal of homodimers pose great challenges to analytical and purification teams, respectively, as these byproducts share high similarity in physicochemical properties with the target heterodimeric bsAb. Protein L is a bacterial surface protein that binds to the variable region of kappa light chain without interfering with the antigen binding site. In this work, we first showed that different antibodies bind Protein L-conjugated resin with varied strength, and then based on this observation we further demonstrated that Protein L chromatography can be a useful tool for monitoring/separating homodimers during the purification of asymmetric bsAbs.
不对称IgG样双特异性抗体（bsAbs）通常源自两种不同来源的亲本单克隆抗体。尽管在设计中实施了促进异源二聚化的策略，但在这些分子的重组生产过程中，仍可能会以较低水平发生同源二聚化。一般来说，监测和去除同源二聚体分别给分析团队和纯化团队带来了巨大挑战，因为这些副产物在物理化学性质上与目标异源二聚体bsAb高度相似。蛋白L是一种细菌表面蛋白，它能与κ轻链的可变区结合而不干扰抗原结合位点。在这项工作中，我们首先表明不同的抗体与蛋白L偶联树脂结合的强度不同，然后基于这一观察结果，我们进一步证明蛋白L色谱法可作为在不对称bsAbs纯化过程中监测/分离同源二聚体的有用工具。
