The typical workflow probably includes front-end enrichment of glycoproteins/glycopeptides, peptide sequence identification, glycosylation sites determination, and glycan structure analysis. CD BioGlyco has professional researchers and experimental instruments, which can provide customers with efficient separation, purification, and analysis services of glycans.
Protein glycosylation analysis can be performed on different levels, including intact glycoproteins, glycopeptides, and released glycans. The analysis of complete glycoprotein is difficult to achieve good separation of different glycoforms of glycoproteins, especially for proteins with a variety of glycosylation sites. Glycopeptide analysis can realize the allocation of glycosylation to specific positions on proteins, and this site-specific information can be used to assign specific glycan structures to different glycosylation sites. In addition, it is helpful to understand the molecular structure of proteins.
Glycans can be released from proteins by chemical or enzymatic methods, and then derivatization steps are usually performed to improve the properties of glycans for analysis. In addition, when dealing with complex biological samples, glycans enrichment need to be performed. Lectin enrichment, graphitized carbon chromatography, hydrophilic liquid chromatography (HILIC), and reverse-phase solid phase extraction are the most commonly used methods for glycan enrichment. Separation techniques such as capillary electrophoresis (CE) are usually used in combination with mass spectrometry (MS) or fluorescence detection. For the analysis of released glycans, various LC stationary phases including HILIC, porous graphitized carbon (PGC), and reversed-phase stationary phase were used.
Fig.1 Strategy workflow for systematic study of N-glycoproteins. (Ruiz-May, 2014)
CD BioGlyco has an advanced and systematic platform for glycans separation, purification, and structure analysis. Currently, we have cooperated with thousands of enterprise customers and provided professional technical services for them. We are confident to provide customers with characteristic and efficient detection solutions. If you have a demand for glycan separation and purification, please contact us for more detailed information.
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