Monosaccharide profiling of glycoproteins by capillary electrophoresis with contactless conductivity detection

Abstract

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.

Triazoles as a Potential Threat to the Nutritional Quality of Tomato Fruits

“…The content of glucose, fructose and saccharose in freeze-dried and peeled tomato fruits was determined by capillary electrophoresis with capacitively coupled contactless conductivity detection on the G7100A Capillary Electrophoresis Instrument (Agilent Technologies, Waldbronn, Germany) [ 43 ].…”

Research of saccharides and related biocomplexes: A review with recent techniques and applications

“…Better utilization of wood waste was shown to improve the production of monosaccharides, polysaccharides, and saccharide metabolites [30,43,201,207,284]. However, the purification of individual saccharides needs large-scale isolation processes Saccharides of aerosols hold valuable information about the sources of organic carbons and their drift, which have implications for human health [32,192,213,214,218,[308][309][310][311][312]. Levoglucosan is a tracer of biomass burning in aerosols [32,213].…”

“…Fetuins are blood proteins that are made in the liver and secreted into the bloodstream. In addition, CE‐CCD4 could also be used as a pre‐concentration technique and profiling tool in glycoproteomics and a complement to the routinely used LC‐MS/MS [311].…”

Establishment and application of characteristic degradation fingerprint for the quality control of Compound Banlangen Granules polysaccharides

“…Compared with ultraviolet spectroscopy, infrared spectroscopy, and high-performance liquid chromatography, capillary electrophoresis has the advantages of high column efficiency, large peak capacity, and simple sample pretreatment. For many years, capillary electrophoresis has been widely used in sugar analysis [18][19][20][21][22]. Zhang et al [23] used glycation enzyme to remove the excipient dextrin in Banlangen Granules, established a fingerprint of partial degradation products of polysaccharide in Banlangen Granules by capillary zone electrophoresis (CZE), and evaluated the quality of preparations from different manufacturers on the basis of polysaccharides and its partial degradation products by cluster analysis.…”

“…Compared with ultraviolet spectroscopy, infrared spectroscopy, and high‐performance liquid chromatography, capillary electrophoresis has the advantages of high column efficiency, large peak capacity, and simple sample pretreatment. For many years, capillary electrophoresis has been widely used in sugar analysis [18–22]. Zhang et al.…”