化学
抛光
离子色谱法
离子交换
洗脱
色谱法
亲和层析
重组DNA
产量(工程)
配对
生物物理学
离子
生物化学
酶
有机化学
基因
材料科学
超导电性
物理
量子力学
生物
冶金
复合材料
作者
Xiaoying Liang,Qingquan He,Guohong Qin,Guozhu Li,Qian Li,Huanghong Tan,Zichen Wang,Minghui Fan,Dan Xu
标识
DOI:10.1016/j.jchromb.2023.123767
摘要
Small amounts of by-products are nevertheless created during the recombinant production of IgG-like bispecific antibodies due to imbalanced chain expression and improper chain pairing, despite the employment of molecular strategy techniques to promote accurate pairing. Among them, homodimers represent the species that are more difficult to remove due to their physical and chemical properties being similar to the target antibody. Homodimer by-products are always produced even though various technologies can significantly increase the expression of heterodimers, so a robust purification process to recover high-purity heterodimers is required. Most of the chromatography methods commonly adopt the bind-and-elute mode or two-step to separate homodimers, which has numerous drawbacks such as prolonged process times and limited dynamic binding capacity. Flow-through mode of anion exchange is a frequently-used polishing step for antibodies, but it is typically regarded as being more effective for host-cell protein or host-cell DNA removal rather than other product-related impurities such as homodimers and aggregates. This paper demonstrated that single-step anion exchange chromatography allows high capacity and effective clearance of the homodimer byproduct to be simultaneously achieved, suggesting that weak partitioning was a better polishing strategy for achieving a high level of heterodimer purity. And robust operation range of anion exchange chromatography steps for homodimer removal was also developed by leveraging the design of experiments.
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