山西医科大学学报2024，Vol.55Issue(9)：1219-1227,9.DOI:10.13753/j.issn.1007-6611.2024.09.017
靶向BCMA/CD3的双特异性抗体药物的质量控制研究
Study on quality control of anti-BCMA/CD3 bispecific monoclonal antibody drugs
摘要
Abstract
Objective To establish a critical quality attribute(CQA)quality control(QC)method for bispecific antibody(bsAb)targeting B cell maturation antigen(BCMA)and cluster of differentiation 3(CD3).Method QC of anti-BCMA/CD3 bsAb were inves-tigated in terms of identification,purity,charge heterogeneity,product related impurities,post-translational modifications,and biological activity.The peptide map method using Lys-C enzyme digestion and reverse phase high performance liquid chromatography(RP-HPLC)was used to specifically identify the bsAb.Purity control was performed using size exclusion chromatography(SEC)and reduced/non-reduced sodium dodecyl sulfate capillary electrophoresis(CE-SDS).Ion exchange chromatography(IEC)was used to measure charge heterogeneity and product related impurities.Multi-attribute method(MAM)composing of reverse phase ultra high performance liquid chromatography(RP-UHPLC)and online mass spectrometry was used to monitor post-translational modifications(PTMs).The biological activity was determined using T cell activation and reporter gene assay(RGA).Results The key peptide segments were effectively identified by the peptide map method which benefits for specific identification.The main components,aggregates and frag-ments were effectively distinguished by both SEC and CE-SDS methods to control the purity of the active ingredients.The acid-base variants,target bsAb,and parental residual homodimers were also effectively distinguished by IEC method,which is beneficial for controlling product related impurities.The PTMs,such as aspartic acid isomerization and asparagine deamidation,were detected by MAM method to serve as a powerful supplement to CQA control.The biological activity was evaluated by T cell activation with RGA method.Conclusion Based on the physicochemical and biological characteristics of anti-BCMA/CD3 bsAb,a QC method for their CQAs has been studied and established to ensure the clinical safety and effectiveness of bsAb products from a quality controllable per-spective.关键词
BCMA/CD3/双特异性抗体/多发性骨髓瘤/关键质量属性/多属性监测方法Key words
BCMA/CD3/bispecific antibodies/multiple myeloma/critical quality attributes/multi-attribute method分类
医药卫生引用本文复制引用
杜加亮,于传飞,武刚,段茂芹,李萌,倪永波,崔永霏,郭璐韵,梅玉婷,王兰..靶向BCMA/CD3的双特异性抗体药物的质量控制研究[J].山西医科大学学报,2024,55(9):1219-1227,9.基金项目
药品监管科学全国重点实验室课题(2024SKLDRS0204,2023SKLDRS0112) （2024SKLDRS0204,2023SKLDRS0112）
