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Glycan profiling of monoclonal antibodies using zwitterionic-type hydrophilic interaction chromatography coupled with electrospray ionization mass spectrometry detection

Citation

Mauko, L and Nordborg, A and Hutchinson, JP and Lacher, NA and Hilder, EF and Haddad, PR, Glycan profiling of monoclonal antibodies using zwitterionic-type hydrophilic interaction chromatography coupled with electrospray ionization mass spectrometry detection, Analytical Biochemistry: Methods in The Biological Sciences, 408, (2) pp. 235-241. ISSN 0003-2697 (2011) [Refereed Article]


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DOI: doi:10.1016/j.ab.2010.09.035

Abstract

We present a new method for the analysis of glycans enzymatically released from monoclonal antibodies (MAbs) employing a zwitterionic-type hydrophilic interaction chromatography (ZIC-HILIC) column coupled with electrospray ionization mass spectrometry (ESI-MS). Both native and reduced glycans were analyzed, and the developed procedure was compared with a standard HILIC procedure used in the pharmaceutical industry whereby fluorescent-labeled glycans are analyzed using a TSK Amide-80 column coupled with fluorescence detection. The separation of isobaric alditol oligosaccharides present in monoclonal antibodies and ribonuclease B is demonstrated, and ZIC-HILIC is shown to have good capability for structural recognition. Glycan profiles obtained with the ZIC-HILIC column and ESI-MS provided detailed information on MAb glycosylation, including identification of some less abundant glycan species, and are consistent with the profiles generated with the standard procedure. This new ZIC-HILIC method offers a simpler and faster approach for glycosylation analysis of therapeutic antibodies.

Item Details

Item Type:Refereed Article
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Separation Science
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Chemical Sciences
Author:Mauko, L (Ms Lea Mauko)
Author:Nordborg, A (Dr Anna Nordborg)
Author:Hutchinson, JP (Dr Joseph Hutchinson)
Author:Hilder, EF (Professor Emily Hilder)
Author:Haddad, PR (Professor Paul Haddad)
ID Code:72206
Year Published:2011
Web of Science® Times Cited:29
Deposited By:Chemistry
Deposited On:2011-08-24
Last Modified:2017-10-30
Downloads:3 View Download Statistics

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