Comparison of traditional 2-AB Fluorescence LC-MS/MS and automated microscale LC-MS for the comparative glycan analysis of monoclonal antibodies

Published

June 5, 2015

Author(s)

John E. Schiel, Michael T. Boyne, Sarah M. Rogstad

Abstract

Monoclonal antibody therapeutics are a heterogeneous mixture of glycoforms. Multiple methods exist for defining the glycan composition and relative abundance of species present. In the current report, two MS-based methods were compared for their ability to both identify glycans, as well as, monitor differences in the glycoprofile as may be useful in characterizing products. The current data indicates a disparity in glycosylation between the analyzed batches of US and Foreign sourced mAb X. Major differences in the glycopofile can most easily be identified by visual inspection of fluorescence profiles, and correlated to glycan identities when coupled with online MS/MS. In comparison, both the LC-F-MS/MS and an automated LC-MS label free approach were able to identify minor differences in low abundance glycoforms, and could serve as a potential screening test for understanding process, comparability, similarity, and possibly detecting counterfeit agents.

Citation

Journal of Pharmaceutical Sciences

Pub Type

Journals

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DOI Link

Keywords

glycan analysis, biopharmaceutical, 2-aminobenzamide, mass spectrometry, fluorescence, comparability, similarity, HILIC

Citation

Schiel, J. , Boyne, M. and Rogstad, S. (2015), Comparison of traditional 2-AB Fluorescence LC-MS/MS and automated microscale LC-MS for the comparative glycan analysis of monoclonal antibodies, Journal of Pharmaceutical Sciences, [online], https://doi.org/10.1002/jps.24522 (Accessed May 11, 2026)

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