Global characterization of modifications to the charge isomers of IgG antibody

作     者：Xinling Cui Wei Mi Zhishang Hu Xiaoyu Li Bo Meng Xinyuan Zhao Xiaohong Qian Tao Zhu Wantao Ying Xinling Cui;Wei Mi;Zhishang Hu;Xiaoyu Li;Bo Meng;Xinyuan Zhao;Xiaohong Qian;Tao Zhu;Wantao Ying

作者机构：State Key Laboratory of ProteomicsBeijing Proteome Research CenterNational Center for Protein Sciences(Beijing)Beijing Institute of LifeomicsBeijingChina National Institute of MetrologyCenter for Advanced Measurement ScienceBeijingChina National Institute of MetrologyChemical Metrology&Analytical Science DivisionBeijingChina CanSino Bio Corporation Tianjin Key Laboratory of Recombinant Bacterial Recombination and Bacterial VaccinesTianjinChina 

出 版 物：《Journal of Pharmaceutical Analysis》 (药物分析学报（英文版）)

年 卷 期：2022年第12卷第1期

页      面：156-163页

核心收录：

学科分类：1001[医学-基础医学(可授医学、理学学位)] 100102[医学-免疫学] 10[医学] 

基　　金：the financial support from the National Key Program for Basic Research of China(Grant Nos.:2018YFC0910302 and 2017YFF0205400) the National Natural Science Foundation of China(Grant No.:81530021) Innovation Foundation of Medicine(Grant Nos.:BWS14J052 and 16CXZ027) 

主　　题：Antibody Charge isomers Mass spectrometry Posttranslational modification Glycopeptide 

摘      要：Posttranslational modifications of antibody products affect their stability,charge distribution,and drug activity and are thus a critical quality *** comprehensive mapping of antibody modifications and different charge isomers(CIs)is of utmost importance,but is *** intended to quantitatively characterize the posttranslational modification status of CIs of antibody drugs and explore the impact of posttranslational modifications on charge *** CIs of antibodies were fractionated by strong cation exchange chromatography and verified by capillary isoelectric focusing-whole column imaging detection,followed by stepwise structural characterization at three ***,the differences between CIs were explored at the intact protein level using a top-down mass spectrometry approach;this showed differences in glycoforms and deamidation ***,at the peptide level,common modifications of oxidation,deamidation,and glycosylation were *** mapping showed nonuniform deamidation and glycoform distribution among *** total,10 N-glycoforms were detected by peptide ***,an in-depth analysis of glycan variants of CIs was performed through the detection of enriched *** and quantitative analyses demonstrated the dynamics of 24 *** results revealed that sialic acid modification is a critical factor accounting for charge heterogeneity,which is otherwise missed in peptide mapping and intact molecular weight *** study demonstrated the importance of the comprehensive analyses of antibody CIs and provides a reference method for the quality control of biopharmaceutical analysis.