Saccharides form one of the major constituents of biological macromolecules in living organisms. Many biological processes including protein folding, stability, immune response and receptor activation are regulated by glycosylation. In this work, we optimized a capillary electrophoresis method with capacitively coupled contactless conductivity detection for the separation of eight monosaccharides commonly found in glycoproteins, namely D-glucose, D-galactose, D-mannose, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-fucose, N-acetylneuraminic acid, and D-xylose. A highly alkaline solution of 50 mM sodium hydroxide, 22.5 mM disodium phosphate, and 0.2 mM CTAB (pH 12.4) was used as a background electrolyte in a 10 μm id capillary. To achieve baseline separation of all analytes, a counter-directional pressure of -270 kPa was applied during the separation. The limits of detection of our method were below 7 μg/ml (i.e., 1.5 pg or 1 mg/g protein) and the limits of quantification were below 22 μg/ml (i.e., 5 pg or 3 mg/g protein). As a proof of concept of our methodology, we performed an analysis of monosaccharides released from fetuin glycoprotein by acid hydrolysis. The results show that, when combined with an appropriate pre-concentration technique, the developed method can be used as a monosaccharide profiling tool in glycoproteomics and complement the routinely used LC-MS/MS analysis.
- MeSH
- acetylgalaktosamin MeSH
- acetylglukosamin MeSH
- cetrimonium MeSH
- chromatografie kapalinová MeSH
- elektroforéza kapilární metody MeSH
- elektrolyty chemie MeSH
- fetuiny MeSH
- fosfáty MeSH
- fukosa MeSH
- galaktosa MeSH
- glukosa MeSH
- glykoproteiny chemie MeSH
- hydroxid sodný MeSH
- kyselina N-acetylneuraminová * MeSH
- mannosa MeSH
- monosacharidy * analýza MeSH
- tandemová hmotnostní spektrometrie MeSH
- xylosa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
