文献类型：期刊文献

英文题名：Identification of multiple sources of the acidic charge variants in an IgG1 monoclonal antibody

作者：Miao, Shiwei[1];Xie, Panpan[1];Zou, Mao[1];Fan, Li[1];Liu, Xuping[1];Zhou, Yan[1];Zhao, Liang[1];Ding, Ding[2];Wang, Haibin[2];Tan, Wen-Song[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Hisun Pharmaceut Hangzhou Co Ltd, Hangzhou 311404, Zhejiang, Peoples R China

年份：2017

卷号：101

期号：14

起止页码：5627

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20171703605768);WOS:【SCI-EXPANDED(收录号:WOS:000405181100005)】；

基金：This work was supported by the National Natural Science Foundation of China (Nos. 21206040 and 21406066).

语种：英文

外文关键词：mAb IgG1; Acidic charge heterogeneity; Deamidation; Galactosylation; Disulfide bond reduction

摘要：Charge variants, especially acidic charge variants, of recombinant monoclonal antibodies are the major critical quality attributes in the biotechnology industry due to their potential influence on stability and biological activity. The chemical properties of the acidic charge variants have been challenging to fully characterize, and it is critical for process development and optimization. To completely understand the multiple sources of acidic charge variants, the major charge forms of an IgG1 monoclonal antibody were firstly isolated and then analyzed by a battery of characterization tools. It was found that various degrees of disulfide bond reduction, the deamination of HC-T8 Asn84 and HC-T35 Asn388 and aggregation account for the majority of acidic charge heterogeneity and the terminal galactosylation content was in relation to the acidic charge heterogeneity. The correlation between acidic charge heterogeneity and galactosylation content was further explored by weak cation exchange chromatography with the use of beta 1-4 galactosidase digestion. The results showed that galactosylation was not the source of the acidic charge variants per se. Meanwhile, to gain insights into the impact on binding affinity of monoclonal antibody to IgE and FcRn, charge variants were also analyzed by competitive ELISA and surface plasmon resonance, respectively. All isolated charge variants had similar affinity binding to IgE and FcRn binding relative to the starting material.