Online Hydrophobic Interaction Chromatography-Mass Spectrometry for the Analysis of Intact Monoclonal Antibodies

Analytical Chemistry
Bifan ChenYing Ge

Abstract

Therapeutic monoclonal antibodies (mAbs) are an important class of drugs for a wide spectrum of human diseases. Liquid chromatography (LC) coupled to mass spectrometry (MS) is one of the techniques in the forefront for comprehensive characterization of analytical attributes of mAbs. Among various protein chromatography modes, hydrophobic interaction chromatography (HIC) is a popular offline nondenaturing separation technique utilized to purify and analyze mAbs, typically with the use of non-MS-compatible mobile phases. Herein we demonstrate for the first time, the application of direct HIC-MS and HIC-tandem MS (MS/MS) with electron capture dissociation (ECD) for analyzing intact mAbs on quadrupole-time-of-flight (Q-TOF) and Fourier transform ion cyclotron resonance (FTICR) mass spectrometers, respectively. Our method allows for rapid determination of relative hydrophobicity, intact masses, and glycosylation profiles of mAbs as well as sequence and structural characterization of the complementarity-determining regions in an online configuration.

References

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Citations

Sep 8, 2020·Mass Spectrometry Reviews·Trisha Tucholski, Ying Ge
Apr 19, 2019·F1000Research·John R Yates
Sep 18, 2020·Advances in Colloid and Interface Science·Shixiang LiuHailin Cong
Nov 2, 2019·Journal of Pharmaceutical and Biomedical Analysis·Julien CamperiNathalie Delaunay
May 14, 2021·Journal of the American Society for Mass Spectrometry·Jake A MelbyYing Ge
Jun 23, 2021·Current Opinion in Biotechnology·Florian FüsslJonathan Bones
Nov 5, 2020·Analytical Chemistry·Jiangnan ZhengRuijun Tian
Sep 1, 2020·Journal of the American Society for Mass Spectrometry·Yuetian YanNing Li
Nov 2, 2019·Analytical Chemistry·Luis A MaciasJennifer S Brodbelt
Sep 1, 2021·Bioanalysis·Shulei Liu, Benjamin L Schulz

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