Study on quality control of anti-BCMA/CD3 bispecific monoclonal antibody drugs
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.
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