Glycosylation Protein Analysis Service

Introduction

Glycosylation is a common and important post-translational modification that plays an important role in various biological processes, including intracellular and intercellular signaling, inflammation, tumor immunology, protein folding, and protein stabilization. Protein glycosylation, that is, the attachment of sugar moieties to proteins, mainly includes N-linked glycosylation (N-glycosylation) and O-linked glycosylation (O-glycosylation). It is estimated that at least 50% of human proteins are glycosylated. However, only about 10% of proteins in existing databases are annotated as glycoproteins, and abnormal protein glycosylation and glycan alterations are associated with many diseases. Therefore, comprehensive analysis of protein glycosylation with high sensitivity is helpful for early detection of diseases or evaluation of disease treatment effects.

Fig. 1. Major types of glycosylation in humans. (Reily C, et al., 2019)

Our Services

Creative Proteomics has launched an advanced protein glycosylation analysis platform with a powerful mass spectrometry platform, and combined with nano LC-MS/MS, on the basis of rich experience in mass spectrometry sequencing.

At Creative Proteomics, our experienced scientists utilize the HCD/ETD "double fragmentation mode" of Orbitrap Fusion mass spectrometry to resolve the complete glycopeptides of complex glycoproteins, and combines the bioinformatics Byonic analysis software to analyze the mass spectrometry original data analysis, realized the identification of N-glycosylation sites and O-glycosylation sites, and analyzed the sugar chain composition carried by the corresponding sites. In addition, the quantitative analysis of glycoproteins was achieved by nano LC-MS/MS.

According the nature of your glycosylated protein, our high-quality scientists will select a specific method, including trypsin digestion or chemical means, to release glycans from glycoproteins and to enrich glycopeptides due to the high structural complexity of glycosylation, which avoids the disadvantage that mixtures of peptides and glycopeptides complicate the analysis.

With cutting-edge equipment and expertise in protein sequencing and mass spectrometry, we provide one-stop glycosylation protein analysis service. You only need to tell us the purpose of your experiment and send your samples to us, we will take care of all the follow-up matters of the project.

Our simple analysis process includes:

• Protein extraction.
• Glycan release.
• Specific enzymatic proteolysis (usually with trypsin).
• Glycosylated peptide enrichment.
• Peptide fragmentation separation.
• Mass spectrometry analysis.
• Bioinformatics analysis.

What Can We Offer You?

• Glycosylation Peptide Enrichment
• Glycan Structural Characterization
• Glycosylation Site and Glycoform Analysis of This Site
• Glycosylation Site Analysis
• Glycoform Analysis
• Glycoprotein Quantification
• Glycosylation Protein-Protein Interaction Analysis

Advantages of Our Services

• High site coverage: Various chemical methods (mainstream lectin affinity method, hydrophilic interaction chromatography and non-specific protease digestion, etc.) are used to enrich glycosylated peptide before mass spectrometry detection, with high specificity and good enrichment rate.
• High accuracy: The quantitative technology of dimethyl labeling is adopted to improve the quantitative accuracy.
• Bioinformatics: The Byonic software is used to achived qualitative and quantitative analysis of intact N-glycopeptide and O-glycopeptide.
• Experienced: Professional advice from scientists with years of experience in mass spectrometry.
• Fast turnaround: 5-7 days for a complete report.
• Customized service: Tailoring the best solution according to your project and sample type.

Sample Requirements

1. Tissue samples: Plant tissue samples > 200 mg, blood samples > 1 mL (with EDTA for anticoagulation of plasma), serum > 0.5 mL, urine > 2 mL, animal tissue samples >1 g, cell samples > 5X10⁷ cells, microorganisms > 200 mg (dry weigh).
2. Protein samples > 1 mg.

Thanks to our powerful mass spectrometry sequencing platform, Creative Proteomics provides a one-stop shop glycosylation protein analysis service for customers worldwhile. Our experienced scientists work with you to develop tailor-made analytical solutions. Please feel free to contact us with any questions regarding glycosylation protein analysis.

References

1. Reily C, Stewart T J, Renfrow M B, et al. (2019) Glycosylation in health and disease. Nat Rev Nephrol. 15(6):346-366.
2. Banazadeh A, Veillon L, Wooding K M, et al. (2017) Recent advances in mass spectrometric analysis of glycoproteins. Electrophoresis. 38(1):162-189.