Intact Glycopeptide Proteomics Analysis
Intact glycopeptide proteomics is a premium technical service that integrates glycobiology, mass spectrometry analysis, and bioinformatics. Unlike conventional proteomics approaches, this service performs comprehensive identification and quantification of glycopeptides while preserving the intact glycan structures, simultaneously capturing the correspondence between glycan structures, modification sites, and peptide sequences.
Why Choose Intact Glycopeptide Analysis?
More than 50% of human proteins undergo glycosylation modifications. Glycosylation is a critical post-translational modification that alters molecular specificity and affects cellular recognition, adhesion, metabolism, and immune processes, thereby influencing disease initiation and progression. Traditional methods that remove glycans prior to analysis result in loss of essential glycan structural information, leading to inability to distinguish multiple glycan types at the same amino acid site, loss of complex glycan heterogeneity information, and difficulty in establishing connections between glycosylation and biological function. Intact glycopeptide analysis overcomes these limitations, providing comprehensive, accurate, and in-depth glycoproteomics data and represents an essential approach for understanding glycoprotein biological functions and disease mechanisms.
The challenge lies in the fact that the same amino acid site can be modified with multiple glycan types. Traditional methods involving enzymatic removal of glycans followed by analysis of residual peptide backbones are incomplete, making accurate identification of glycopeptides and glycan type determination extremely difficult. Our intact glycopeptide identification technology preserves glycan structures for direct analysis, precisely mapping specific glycan types at each modification site. This breakthrough approach transcends the limitations of conventional methods, delivering more complete and accurate glycoproteomics data.
Figure 1. Overview of the experimental strategy and molecular structure of Infliximab. (Source: Kim H, et al. 2022)
Our Services
Sample Preparation → Glycopeptide Enrichment → Liquid Chromatography Separation → Mass Spectrometry Analysis → Bioinformatics Data Processing
| Analysis Type | Applicable Targets | Data Deliverables |
| N-linked Intact Glycopeptide Analysis | Glycosylation modifications at N-X-Ser/Thr motifs (where X is any amino acid except proline) | Complete N-glycopeptide identification and statistics, glycan structure and modification site heatmaps, glycan distribution and core structure analysis, detailed branch modification dissection, annotated spectra for top 10 abundant glycans |
| O-linked Intact Glycopeptide Analysis | O-type glycosylation modifications on Ser/Thr residues, including GalNAc-initiated mucin-type O-glycosylation, GlcNAc-initiated O-GlcNAc modifications, and other O-type glycosylation | Complete O-glycopeptide sequence and site localization, detailed O-glycan chain structure analysis, site-specific glycan distribution, O-glycopeptide abundance statistics |
| Quantitative Analysis Solutions | Supports two quantification modes | Differential glycopeptide volcano plots and clustering heatmaps, glycoprotein functional annotation (GO/KEGG pathway analysis), protein-protein interaction network analysis, subcellular localization prediction |
Service Advantages
Our intact glycopeptide proteomics service delivers industry-leading detection depth and accuracy. By preserving intact glycan structures for mass spectrometry analysis, we capture complete correspondence information among glycan structures, modification sites, and peptide sequences, achieving single amino acid site resolution where multiple glycan types at the same site can be fully distinguished.
Workflow
Initial communication regarding research objectives, sample types, and expected depth, followed by sample feasibility assessment and development of optimal technical and budget plans tailored to your needs.
Verification of sample quality and quantity, protein quantification, and timely feedback on QC results with recommendations for improvement if samples do not meet requirements.
Following standardized protocols for protein digestion, glycopeptide enrichment, and preprocessing optimization, ensuring each step meets rigorous standards and providing a solid foundation for subsequent analysis.
Using high-resolution mass spectrometry instruments for sample analysis, automatic data acquisition and conversion, continuous quality monitoring, and parameter optimization to ensure data accuracy and completeness.
Performing glycopeptide identification and quantification, statistical analysis and inter-group comparison, and multi-dimensional data visualization to uncover the biological significance underlying the data.
Providing comprehensive analysis reports containing raw data, processed data, and statistical results, with professional interpretation and application recommendations from the technical team.
Reference
- Kim H, et al. Advanced assessment through intact glycopeptide analysis of Infliximab's biologics and biosimilar. Front Mol Biosci. 2022 Nov 29;9:1006866.
