Glycosylation Site Identification Service

MtoZ Biolabs provides Glycosylation Site Identification Service that integrates optimized protease digestion, selective glycopeptide enrichment, and high-resolution LC-MS/MS platforms to deliver precise mapping of N-linked and O-linked glycosylation sites. With workflows tailored to diverse sample types and project objectives, we ensure accurate, and reproducible results that support both academic research and biopharmaceutical development.

 

Overview

Glycosylation Site Identification focuses on determining the exact amino acid residues where glycans are covalently attached. This provides critical site-specific insights into how glycosylation alters protein structure and function. By identifying whether glycosylation occurs, where it occurs, and the type and structure of attached glycans, researchers gain a complete picture of glycoprotein regulation. This information is essential for both fundamental biology, such as understanding cell signaling and protein interactions, and applied research, including biomarker discovery, disease mechanism studies, and vaccine development.

 

With the advancement of high-resolution mass spectrometry and bioinformatics analysis, Glycosylation Site Identification has become a robust and reliable approach for comprehensive glycoprotein characterization. It provides not only localization of N-linked and O-linked glycosylation sites but also quantitative site occupancy data and structural details of glycans. These capabilities are indispensable for biopharmaceutical development, biosimilar comparability, and regulatory compliance.

 

Oh MJ. et al. Mass Spectrom Rev. 2025.

Figure 1. Streamlined Workflow for Site‐Specific Glycopeptide Analysis of Therapeutic Glycoproteins.

  

Services at MtoZ Biolabs

1. Target Proteins Glycosylation Site Identification 

MtoZ Biolabs can perform precise analysis of glycosylation sites on target proteins, identifying specific modified residues and their occupancy. Relying on a high-resolution mass spectrometry platform, we can compare site changes under different conditions, providing clear evidence for studying structural impact and functional regulation.

2. Glycosylation Proteomics Site Identification 

By combining enrichment strategies with large-scale mass spectrometry analysis, MtoZ Biolabs can systematically obtain glycosylation site information for numerous proteins in a sample, constructing an overall modification distribution profile. This analysis helps evaluate glycosylation patterns across different biological states and supports more in-depth regulatory mechanism studies.

 

Analysis Workflow

The general analytical workflow for the Glycosylation Site Identification Service is as follows:

1. Sample Preparation and Quality Control

Proteins from biological or pharmaceutical sources are evaluated for concentration and purity, with buffer exchange or cleanup performed if required.

2. Enzymatic Digestion

Proteins are digested using proteases such as trypsin or complementary enzymes to generate peptides and glycopeptides while maintaining glycosylation integrity.

3. Glycopeptide Enrichment

Enrichment methods including lectin affinity chromatography, HILIC, or hydrazide chemistry are applied to selectively capture glycopeptides and reduce interference from non-glycosylated peptides.

4. LC-MS/MS Analysis

High-resolution mass spectrometry platforms such as Orbitrap and Q-TOF are used with advanced fragmentation techniques (CID, HCD, ETD) to identify peptide sequences and glycosylation sites.

5. Data Processing and Bioinformatics

Spectral data are processed with specialized software and curated glycan databases to confirm glycosylation sites, annotate glycan structures, and quantify site occupancy.

6. Reporting

Clients receive a detailed report including identified N- and O-glycosylation sites, structural information of glycans, quantitative data, and interpretation.

 

Service Advantages

1. Comprehensive Site Mapping

Accurate identification of both N-linked and O-linked glycosylation sites with high confidence, providing site-specific insights into protein regulation.

2. Advanced MS Platforms and Enrichment

Integration of selective glycopeptide enrichment with Orbitrap and Q-TOF LC-MS/MS ensures high sensitivity, structural resolution, and reliable detection of low-abundance glycopeptides.

3. High-Data-Quality

Deep data coverage with strict data quality control. AI-powered bioinformatics platform integrates all Glycosylation Site Identification Service data, providing clients with a comprehensive data report.

  

Sample Submission Suggestions

 

It is recommended to contact our technical support team before submitting samples to determine sample suitability and obtain tailored submission guidelines.

 

Applications

Practical applications of Glycosylation Site Identification Service are as follows:

1. Therapeutic Protein Development

Evaluate glycosylation site occupancy and heterogeneity in monoclonal antibodies, fusion proteins, and therapeutic enzymes to meet regulatory standards.

2. Biosimilar Comparability

Compare glycosylation site profiles between biosimilars and reference products to demonstrate structural and functional equivalence.

3. Vaccine Design

Identify glycosylation sites on viral antigens to understand immunogenicity and stability, guiding vaccine development.

4. Cancer Research

Map site-specific glycosylation alterations associated with tumor initiation, progression, and metastasis.

 

FAQ

Q1: Can you Provide Quantitative Glycosylation Site Occupancy Data?

A1: Yes. We offer relative quantification of glycoforms and occupancy rates at each identified site, enabling comparisons across samples, batches, or experimental conditions.

Q2: Is It Possible to Study Glycosylation in Combination with Other Post-Translational Modifications?

A2: Yes. Our workflows can be integrated with phosphoproteomics, acetylomics, or ubiquitinomics to investigate crosstalk between glycosylation and other PTMs, offering a more complete view of protein regulation.