Investigation of the correlation between charge and glycosylation of IgG1 variants by liquid chromatography–mass spectrometry |
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Authors: | Jia-Ming Yang Junwen AiYuemei Bao Zhijun YuanYumin Qin Yi-Wu XieDesheng Tao Daotian FuYucai Peng |
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Institution: | Livzon MabPharm, Zhuhai, Guangdong 519020, China |
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Abstract: | A recombinant IgG1 monoclonal antibody (mAb) showed multiple charge variants in a cation exchange chromatography profile. To better understand the correlation between charge heterogeneity and glycosylation, a rapid reversed phase ultra-performance liquid chromatography–mass spectrometry (UPLC–MS) method with integrated mass analysis has been developed for simultaneous determination of N-terminal pyroglutamate, C-terminal lysine truncation, and Fc glycosylation. The results show that various degrees and/or types of N-terminal pyroglutamate formation and C-terminal lysine (Lys) cleavage account for the majority of charge heterogeneity; and the charge variants showed Fc glycosylation patterns in relation to their terminal modifications. The amount of G1F decreased in the basic variants, whereas Man5 and G0F-GN increased. The complement-dependent cytotoxicity (CDC) activity of purified charge variants also suggested the potential impact of the charge differences on the glycosylation profile. |
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Keywords: | Monoclonal antibody UPLC&ndash MS Charge heterogeneity Glycosylation |
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