治疗性单克隆抗体电荷异质性分析方法比较

发布时间：2018-03-29 02:16

本文选题：单克隆抗体　切入点：电荷异质性　出处：《药物分析杂志》2017年08期

【摘要】：目的:对4种常用的单抗电荷异质性分析方法进行比对研究。方法:以重组抗TNFα全人源单克隆抗体为例,分别利用离子交换色谱(IEC)、毛细管区带电泳(CZE)、毛细管等电聚焦电泳(CIEF)和成像毛细管等电聚焦电泳(iCIEF)4种方法对该抗体的电荷异质性进行分析;利用CIEF和iCIEF 2种方法对抗体主峰等电点(pI)进行分析。结果:4种方法能够对重组抗TNFα全人源单克隆抗体的电荷异构体进行有效的分离和定量,其中IEC和CZE 2种方法检测的电荷异构体峰面积百分比(酸性峰、主峰和碱性峰)较为一致,4种方法检测的碱性峰比例较一致,CIEF和iCIEF检测的主峰与酸性峰比例较其他方法偏高。CIEF和iCIEF检测抗体主峰等电点具有一定差异,其中,聚焦时间长短和pI Marker的选择是影响单抗pI值检测的主要因素。结论:通过抗体电荷异质性分析方法的比较研究可见,在单抗电荷异构体比例和主峰pI测定上4种方法具有一定的差异。对单抗的质控应结合产品特性和表征研究,合理选择电荷异质性分析方法。
[Abstract]:Objective: to compare and study four common methods for the analysis of the charge heterogeneity of monoclonal antibodies (McAbs). Methods: the recombinant monoclonal antibody against TNF 伪 was used as an example. The charge heterogeneity of the antibody was analyzed by ion-exchange chromatography (IEC), capillary zone electrophoresis (CZE), capillary isoelectric focusing electrophoresis (CIEF) and imaging capillary isoelectric focusing electrophoresis (iCIEF). The main peak isoelectric point (Pi) of the antibody was analyzed by CIEF and iCIEF. Results the charge-isomers of the recombinant anti-human monoclonal antibody against TNF 伪 could be effectively separated and quantified by the two methods. The charge isomer peak area percentage detected by IEC and CZE methods (acid peak, acid peak, acid peak, acid peak, acid peak, acid peak, acid peak, acid peak). The ratio of main peak and acid peak detected by iCIEF and iCIEF were higher than those of other methods, and the isoelectric point of main peak of antibody detected by iCIEF was higher than that of other methods, among them, the ratio of main peak to acid peak was higher than that of other methods, and the ratio of main peak to acid peak was higher than that of other methods. The length of focus time and the selection of Pi Marker are the main factors influencing the detection of Pi value of monoclonal antibody. Conclusion: through the comparison of antibody charge heterogeneity analysis, we can see that, There are some differences in the ratio of charge isomers of McAb and the determination of main peak Pi. The quality control of McAbs should be combined with the study of product characteristics and characterization, and the analysis method of charge heterogeneity should be reasonably selected.
【作者单位】：中国食品药品检定研究院单克隆抗体产品室卫生部生物技术产品检定及标准化重点实验室;
【基金】：国家“重大新药创制”科技重大专项资助项目(2014ZX09304311-001,2012ZX09304010)
【分类号】：R392-33

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