Protein glycosylation is ubiquitous and important in biological systems. Among hundreds of known modifications, glycosylation is one of the most common and necessary modifications. It determines protein folding, transportation and stability, and regulates almost all cell activities, including gene expression, signal transduction, extracellular activities and other responses. Glycoproteomics will remain an attractive research area because of the realization that glycosylation may be involved in the progress of many diseases. Therefore, the development of effective analytical tools to detect abnormal glycoprotein is very important for the in-depth understanding of the pathological function of glycoproteins and the development of new biomarkers. It is difficult to systematically and comprehensively analyze glycosylation modifications with conventional biochemical methods, but mass spectrometry in recent years has provided researchers with unique and site-specific opportunities to characterize protein modifications.
However, glycoproteins are not abundant compared to unmodified proteins. The heterogeneity of its glycan structure and the complexity of the various amino acid residues covalently bound to glycans make it still a great challenge to comprehensive analysis. Therefore, effective enrichment of glycopeptides /glycoproteins is essential before using MS for systematical analysis. Enrichment allows us to minimize the interference of high-abundance non-glycoproteins on glycosylation analysis and obtain enriched glycoproteins. CD BioGlyco provides a variety of advanced methods to enrich glycopeptides/glycoproteins.
Fig 1. Illustration of the most commonly used strategies for glycoproteins/glycopeptides enrichment (Zhao, M.; et al. 2014)
CD BioGlyco provides fast and efficient glycoprotein enrichment services, such as O-Glycoprotein and Peptide Enrichment. Our enrichment methods include but are not limited to:
The glycan is oxidized to generate aldehyde groups, and the oxidized glycopeptide is conjugated to a solid support using hydrazide chemistry. After on-beads digestion, non-glycopeptides can be removed, and N-glycans are cleaved from the beads by using Peptide-N-Glycosidase F to achieve the purpose of enrichment.
Generally, lectins can be immobilized on a solid support for solid phase extraction of glycopeptides. Beads containing lectins can also be packed into separation columns for use in combination with liquid chromatography.
The stationary phase and glycans are highly hydrophilic, so the retention time of glycopeptides on HILIC columns is usually longer than that of non-glycopeptides.
The covalent interactions between boronic acids (BA) and cisdiols on glycans have been extensively studied and been applied for glycoprotein enrichment and analysis.
An isotope target glycoproteomics method that combines metabolic labeling, isotope recording and MS-based proteomics. This innovative method can be used to enrich and analyze intact N- and O- glycopeptides.
CD BioGlyco has many advanced technologies for glycoprotein purification, we guarantee to provide the target glycoprotein that has high purity and high biological activity. Our researchers can also customize the experimental program for you according to your research needs.
Customers can contact our employees directly and we will respond promptly. If you are interested in our services, please contact us for more detailed information.
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