Glycan profiling of monoclonal antibodies using zwitterionic-type hydrophilic interaction chromatography coupled with electrospray ionization mass spectrometry detection
| Autor: | Joseph P. Hutchinson, Nathan A. Lacher, Emily F. Hilder, Paul R. Haddad, Lea Mauko, Anna Nordborg |
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| Přispěvatelé: | Mauko, Lea, Nordborg, Anna, Hutchinson, Joseph P, Lacher, Nathan A, Hilder, Emily F, Haddad, Paul R |
| Rok vydání: | 2010 |
| Předmět: |
Glycan
Spectrometry Mass Electrospray Ionization animal structures Glycosylation medicine.drug_class Electrospray ionization Molecular Sequence Data Biophysics Oligosaccharides Mass spectrometry Monoclonal antibody Biochemistry chemistry.chemical_compound Ribonucleases medicine liquid chromatography Molecular Biology ZIC-HILIC Fluorescent Dyes Chromatography biology Hydrophilic interaction chromatography ESI-MS Antibodies Monoclonal Cell Biology Fluorescence carbohydrates (lipids) hydrophilic interaction chromatography chemistry Carbohydrate Sequence Glycan profiling biology.protein Chromatography Gel glycans monoclonal antibodies Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Analytical biochemistry. 408(2) |
| ISSN: | 1096-0309 |
| Popis: | 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. Refereed/Peer-reviewed |
| Databáze: | OpenAIRE |
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