Site Occupation
Glycosylation is one of the most common protein post-translational modifications. At least 50% of human proteins are glycosylated. N-, O-linked glycans are required to maintain the biological functions of proteins. Insufficient glycosylation can lead to animal and plant diseases. Therefore, characterizing the degree of glycosylation on proteins is an important step in understanding the diagnosis and treatment of diseases. N-, O-glycosylation is also closely related to various biological events, including cancer metastasis, virus infection and antibody-antigen interactions, so the accurate determination of site occupancy is essential to fully understand the impact of protein glycosylation on human health. Unlike N-glycosylation, it is more difficult to analyze O-linked glycans due to the lack of universal enzymes for releasing O-glycans from proteins and the high degree of heterogeneity in their occupation.
Glycoprotein analysis requires the determination of glycosylation sites and the glycan structure associated with each site. In recent years, a wide range of analysis methods based on mass spectrometry have been developed. These methods require high resolution and sensitivity so that glycosylation sites and micro-heterogeneity information can be obtained.
Through high-precision mass spectrometry (MS) combined with bioinformatics analysis, we provide quantitative assessment services of the actual occupancy of glycosylation sites in target proteins, helping clients to reveal the degree of modification of each potential glycosylation site.
CD BioGlyco provides advanced N-glycosylation site occupation analysis services based on MS, usually including labeling, label-free, and multiple reaction monitoring methods.
CD BioGlyco has first-class O-glycosylation analysis platforms and a complete database, which provide clients with comprehensive and time-saving glycosylation analysis.
Our service process includes sample pre-treatment (e.g., enzymatic digestion, glycan release, and enrichment), MS data acquisition, glycosylation site identification, site occupancy calculation (i.e., the ratio of modified sites to total sites), and further analysis of glyco-heterogeneity, glycan structure, and dynamic changes.
Technology: Immunoradiometric assay (IRMA), Pharmacokinetic studies, Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)
Journal: International Journal of Molecular Sciences
Published: 2017
IF: 4.9
Results: This review applies N-glycosylation profiling techniques to systematically compare the glycosylation site occupancy, glycan structural characteristics, and bioactivity differences between two natural pituitary-derived human thyroid-stimulating hormones (hTSH) and three recombinant hTSH preparations. The recombinant preparations were found to have significantly higher glycosylation occupancy (65%-87%) and glycan mass (12%-19%) than the natural hormones, and the average glycan mass was reduced by 24%. This study provides an important basis for understanding the effects of glycosylation modifications on hTSH function and quality control of recombinant biopharmaceuticals.
Fig.1 MALDI-TOF-MS data of samples. (Ribela, et al., 2017)
CD BioGlyco is an experienced company in the analysis of glycosylation site occupation. Our multi-functional analysis platform and powerful database can provide customers with a diverse portfolio of glycosylation analysis services, and our excellent services have helped us gain a good reputation worldwide. We hope to start the exploration of glycobiology with you.
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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