Simultaneous monitoring of oxidation, deamidation, isomerization, and glycosylation of monoclonal antibodies by liquid chromatography-mass spectrometry method with ultrafast tryptic digestion

MAbs
Yi WangXiaoyu Yang

Abstract

Monoclonal antibodies are subjected to a wide variety of post-translational modifications (PTMs) that cause structural heterogeneity. Characterization and control of these modifications or quality attributes are critical to ensure antibody quality and to define any potential effects on the ultimate safety and potency of antibody therapeutics. The biopharmaceutical industry currently uses numerous tools to analyze these quality attributes individually, which requires substantial time and resources. Here, we report a simple and ultrafast bottom-up liquid chromatography-mass spectrometry (uLC-MS) method with 5 min tryptic digestion to simultaneously analyze multiple modifications, including oxidation, deamidation, isomerization, glycation, glycosylation, and N-terminal pyro-glutamate formation, which can occur during antibody production in mammalian cell culture, during purification and/or on storage. Compared to commonly used preparation procedures, this uLC-MS method eliminates assay artifacts of falsely-increased Met oxidation, Asp isomerization, and Asn deamidation, a problem associated with long digestion times in conventional LC-MS methods. This simple, low artifact multi-attribute uLC-MS method can be used to quickly and ac...Continue Reading

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Citations

Jan 21, 2017·MAbs·Izabela SokolowskaPing Hu
Dec 30, 2017·Expert Opinion on Biological Therapy·William C LamannaMartin Schiestl
Apr 26, 2018·Expert Review of Proteomics·Deepali RathoreSarah Rogstad
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May 3, 2019·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Amita Datta-Mannan
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Sep 19, 2020·Biotechnology and Bioengineering·Dhanuka P WasalathanthriZheng Jian Li
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Jan 30, 2021·Journal of the American Society for Mass Spectrometry·Craig JakesJonathan Bones
Feb 6, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yvonne Ehwang SongUdo Roth
Feb 15, 2020·Journal of Pharmaceutical and Biomedical Analysis·Dongdong WangHongcheng Liu
Jun 22, 2021·Journal of the American Society for Mass Spectrometry·Johanna ParisPeter B O'Connor

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Methods Mentioned

BETA
glycosylation
ion exchange chromatography
deamidation

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