Glycoprotein Quantification
Protein glycosylation is one of the major post-translational modifications that can significantly impact protein folding, distribution, stability, conformation, and activity. Glycoprotein abundance, glycosylation degree, and glycan structure changes are closely related to many diseases (such as genetic diseases, immunodeficiency, cardiovascular diseases, and cancer). The research on glycoproteomics has become popular, increasing the demand for the development of quantitative glycomics strategies. These strategies need to be able to sensitively monitor the abundance and structural changes of glycans. The role and characteristics of glycans in biological systems can be clarified using these strategies.
In the past, people focused their research on the enrichment and separation of glycoproteins/glycopeptides, identification of glycosylation sites, and glycan structure analysis. So far, the development of quantitative glycomics has lagged slightly behind other studies. At present, a combination of multiple analytical techniques (such as capillary electrophoresis, liquid chromatography, and mass spectrometry) is mainly used to analyze glycoprotein quantitatively. To enhance the sensitivity in spectroscopy and mass spectrometry, it is necessary to label with ionic and/or hydrophobic reagents during the analysis. Labeling with stable isotope reagents has also been considered a feasible strategy to achieve relative quantification.
Fig.1 Quantitative proteomic schematic of serum glycoproteins. (Gao, 2015)
CD BioGlyco provides a variety of technologies for quantitative analysis of glycoproteins, mainly including fluorescence spectroscopy, capillary electrophoresis, laser-induced fluorescence detection, hydrophilic interaction chromatography with fluorophores, and mass spectrometry (MS). Our researchers provide the best experiment design according to our client's research needs.
We provide relative quantification of glycoproteins through strategies that combine isotope labeling/isotope-free labeling with a variety of advanced MS techniques. Our professional researchers provide relative quantitative analysis of different levels of glycoproteins.
We have established several absolute quantification methods based on the characteristics of glycoproteins to provide technical support for our clients' scientific research. Our researchers use advanced techniques such as liquid chromatography-mass spectrometry (LC-MS) in combination with stable isotope labeling or standard curve methods to specifically identify and quantify glycoproteins.
Technology: High-performance anion-exchange chromatography (HPAEC), High-performance liquid chromatography (HPLC), Enzyme-linked immunosorbent assay (ELISA)
Journal: International Dairy Journal
Published: 2020
IF: 3.1
Results: This paper presents a method for the quantification of bovine whey glycoproteins based on N-glycosylation profiling. The researchers achieved simultaneous quantification of three major glycoproteins, lactoferrin, IgG, and GlyCAM-1, in whey by optimizing the sample preparation process and chromatographic analysis technique. The researchers revealed the differences in glycoprotein composition in whey powder under different processing conditions, and also verified the consistency of the method with ELISA results, providing an efficient analytical tool for quality control and functional studies of glycoproteins in the dairy industry.
Fig.2 Overall for bovine acid whey glycoprofile. (Valk-Weeber, et al., 2020)
CD BioGlyco is a company with rich experience and a good reputation in the field of glycoproteomics. Our multi-platform methods and well-trained researchers will greatly accelerate our clients' research and help them explore glycobiology more comprehensively and accurately. If you are interested in our services, please contact us for more consultation and we will respond promptly.
We provide comprehensive N-glycosylation site occupancy and O-glycosylation site occupancy analysis services to help resolve the core features of protein glycosylation modifications. Through MS combined with a glycopeptide enrichment strategy, we precisely locate the modification sites of N-glycosylation and O-glycosylation in proteins, and quantitatively assess their site occupancy, revealing the efficiency of linking glycan and peptide chains as well as their heterogeneity.
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