两种阴离子交换层析填料去除重组全人源抗程序性死亡配体1单克隆抗体宿主蛋白能力及动态载量的比较

Host cell protein removal and dynamic binding capacities of two anion exchange chromatographic media used for recombinant human complete monoclonal antibody against programmed cell death-1

目的比较两种阴离子交换层析填料Capto Q和Eshmuno Q去除重组全人源抗程序性死亡配体1(programmed cell death-ligand 1,PD-L1)单克隆抗体宿主蛋白(host cell protein,HCP)的能力以及动态载量。方法采用AKTA avant 25层析系统,通过实验设计(design of experiment,Do E)考查上样量、pH和电导率3个工艺参数变量对HCP去除的影响,设计条件为上样量范围40~60 mg IgG/m L gel,pH范围6.6~7.4,电导率范围1~4 ms/cm,阴离子交换层析采用抗体穿透模式。结果在设计的工艺参数范围内,随着pH的增加和电导率的减小,HCP去除效果有上升趋势;随着pH的减小和电导率的增加,动态载量有上升趋势。在最佳操作范围内当pH=7.0,电导率=1 ms/cm时,Capto Q的HCP去除能力为0.01%,载量为45 mg/m L gel;Eshmuno Q的HCP去除能力为0.02%,载量为35 mg/m L gel。结论阴离子交换层析能有效去除PD-L1单抗的HCP,去除率达50倍以上,动态载量Eshmuno Q达35 mg/m L gel,Capto Q达45 mg/m L gel。

Objective To compare the host cell protein（HCP） removal capability and dynamic binding capacity of two chromatographic media Capto Q and Eshmuno Q used for recombinant human complete monoclonal antibody（Mc Ab）against programmed cell death-1（PD-L1）. Methods The effects of technological parameters such as sample loading volume,pH value and conductivity on removal of HCP were evaluated using AKTA avant 25 system by design of experiment（Do E）. The sample loading volume was designed as 40 ~ 60 mg Ig G/m L gel,while the pH value as 6. 6 to7. 4,the conductivity as 1 ~ 4 ms/cm,and the anion exchange chromatography was performed in antibody penetrating mode. Results Within the range of designed process parameters,the HCP removal capability showed a rising trend as the pH value increased and conductivity decreased,while the dynamic binding capacity increased with the decrease of pH value and the increase of conductivity. However,at the optimal pH value of 7. 0 and the optimal conductivity of1 ms/cm,the HCP removal capacity and sample loading of Copto Q were 0. 01% and 45 mg/m L gel,while those of Eshmuno Q were 0. 02% and 35 mg/m L gel,respectively. Conclusion Anion exchange chromatography effectively removed the HCP in Mc Ab against PD-L1,with an efficiency of more than 50 times,while the dynamic binding capacities of Eshmuno Q and Capto Q were 35 and 45 mg/m L gel respectively.