Department of Biological Chemistry, University of Hasanuddin, Indonesia
Mini Review
Impact of Ion-Pairing Agents on Hydrophilic-Interaction Liquid Chromatography High-Resolution Mass Spectrometry's Ability to Separate Intact Glycoproteins
Author(s): Shiva Mishra*
High-resolution separations of glycoforms of glycoproteins that differ in the number of glycans are possible using hydrophilic-interaction liquid chromatography of intact proteins. To maximise the contribution of the hydroxyl groups of the sugars in the glycoprotein, it is necessary to shield the positively charged sites of the proteins with acidic ion-pair reagents in order to get effective separations. Here, we looked at how different IPRs with diverse physico-chemical characteristics, like hydrophobicity and acidity, affected the capillary-scale HILIC separation of intact. The use of fluoroacetic acid, difluoroacetic acid, trifluoroacetic acid, and heptafluorobutyric acid as diluents for sample preparation, solvents for loading samples onto a reversed-phase trap prior to the HILIC separation, and mobile-phase components for HILIC and HILICMS were all examined. To we used an acrylam.. View More»
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