Characterization of Intact Neo-Glycoproteins by Hydrophilic Interaction Liquid Chromatography
| Autor: | Gabriella Massolini, Petrus Hemström, Marco Terreni, Alice Pedrali, Caterina Temporini, Giorgio Marrubini, Teodora Bavaro, Sara Tengattini |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Glycan Glycosylation Electrospray ionization hydrophilic interaction liquid chromatography Pharmaceutical Science intact glycoproteins semi-synthetic glycosylation monitoring neo-glycoproteins Mass spectrometry High-performance liquid chromatography Article Analytical Chemistry lcsh:QD241-441 chemistry.chemical_compound lcsh:Organic chemistry Drug Discovery Sample preparation Physical and Theoretical Chemistry Acetonitrile Chromatography High Pressure Liquid Glycoproteins Chromatography biology Hydrophilic interaction chromatography Organic Chemistry Ribonuclease Pancreatic chemistry Chemistry (miscellaneous) biology.protein Molecular Medicine Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Molecules; Volume 19; Issue 7; Pages: 9070-9088 Molecules, Vol 19, Iss 7, Pp 9070-9088 (2014) Molecules |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules19079070 |
| Popis: | In this study, an HPLC HILIC-UV method was developed for the analysis of intact neo-glycoproteins. During method development the experimental conditions evaluated involved different HILIC columns (TSKgel Amide-80 and ZIC-pHILIC), and water-acetonitrile mixtures containing various types of acids and salts. The final selected method was based on a TSKgel Amide-80 column and a mobile phase composed of acetonitrile and water both containing 10 mM HClO4. The influence of temperature and sample preparation on the chromatographic performances of the HILIC method was also investigated. The method was applied to the separation of neo-glycoproteins prepared starting from the model protein RNase A by chemical conjugation of different glycans. Using the method here reported it was possible to monitor by UV detection the glycosylation reaction and assess the distribution of neo-glycoprotein isoforms without laborious sample workup prior to analysis. |
| Databáze: | OpenAIRE |
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