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Characterization of disulfide linkages at the hinge region of IgG antibodies by HPLC mass spectrometry
Author(s) -
You Jia,
Shi Ying,
Zhu Wenli,
Wu Zhigang,
Xiong Jingyuan
Publication year - 2018
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4371
Subject(s) - chemistry , disulfide bond , mass spectrometry , antibody , chromatography , disulfide linkage , characterization (materials science) , high performance liquid chromatography , immunoglobulin fc fragments , immunoglobulin g , biochemistry , enzyme , cysteine , nanotechnology , materials science , immunology , biology
There are two types of disulfide linkages in IgG antibodies at the hinge region: intra‐ and inter‐disulfide linkages. Characterization of intra‐disulfide linked isomer will provide important information on the stability of the antibodies and better understanding of the mechanism of Fab‐arm exchange. In this report, HPLC coupled with high‐resolution mass spectrometry was applied for characterization of disulfide linkages in IgG antibodies at the hinge region. We were able to accurately identify both inter‐ and intra‐disulfide linked peptides and correctly quantify intra‐disulfide isomers. It is the first study to quantify intra‐disulfide isomers in IgG antibodies with a mass spectrometry approach. It will help to achieve efficient generation of bispecific antibodies with Fab‐arm exchange.