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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 |
| UTAS Author: | Mauko, L (Ms Lea Mauko) |
| UTAS Author: | Nordborg, A (Dr Anna Nordborg) |
| UTAS Author: | Hutchinson, JP (Dr Joseph Hutchinson) |
| UTAS Author: | Hilder, EF (Professor Emily Hilder) |
| UTAS Author: | Haddad, PR (Professor Paul Haddad) |
| ID Code: | 72206 |
| Year Published: | 2011 |
| Web of Science® Times Cited: | 34 |
| Deposited By: | Chemistry |
| Deposited On: | 2011-08-24 |
| Last Modified: | 2017-10-30 |
| Downloads: | 3 View Download Statistics |
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