Targeted O-Glycosylation Site Mapping Service

MtoZ Biolabs provides specialized Targeted O-Glycosylation Site Mapping Services designed to identify, localize, and quantify O-linked glycosylation sites on specific target proteins. Using advanced LC-MS/MS systems coupled with optimized enzymatic digestion and glycopeptide enrichment strategies, we deliver accurate site-specific O-glycosylation profiles that support protein structure validation, therapeutic development, and quality control.

What Is O-Glycosylation?

Protein O-glycosylation is a post-translational modification in which glycans are attached to the hydroxyl groups of serine or threonine residues. O-glycans are structurally diverse but are mainly classified into eight core structures (Core 1–8) based on their linkage patterns and branching characteristics. These eight core types serve as the foundational backbones for further elongation and modification, generating thousands of O-glycan variants with distinct biological roles.

Watson, M. E. et al. J Gastrointestin Liver Dis. 2015.

Figure 1. O-Glycan Structures Core 1-8

O-Glycosylation Site Mapping refers to the precise identification and localization of O-linked glycosylation sites on a specific protein. This process determines exactly which serine or threonine residues carry glycans and characterizes the structure attached to each site.

O-glycosylation is often heterogeneous and lacks a strict consensus sequence, making site mapping essential for:

🔸Understanding Protein Function: Identifying glycosylation sites helps clarify how these modifications influence protein folding, stability, and biological activity.

🔸Ensuring Product Quality: In therapeutic proteins and antibodies, accurate site mapping verifies structural consistency across production batches.

🔸Supporting Structural Research: Site-specific information provides insights into how O-glycans affect molecular interactions and conformational properties.

Targeted O-Glycosylation Site Mapping Service at MtoZ Biolabs

💠Targeted O-Glycosylation Site Identification: Determine specific O-linked modification sites on proteins using LC-MS/MS with ETD/EThcD fragmentation.

💠Site-Specific Glycan Composition Profiling: Identify the monosaccharide components (GalNAc, Gal, GlcNAc, NeuAc, etc.) at each site.

💠Quantitative Analysis: Evaluate relative glycan occupancy and heterogeneity between protein batches or sample conditions.

💠Structural Verification: Confirm the relationship between glycan composition and protein structure to ensure functional stability.

Workflow of Targeted O-Glycosylation Site Mapping Service

1. Extraction and purification of proteins

2. Digestion of proteins

3. Enrichment and elution of peptides

4. Analysis by mass spectrometry

5. Interpretation of data

Kang, T. et al. Cell Rep Methods. 2024.

Figure 2. A Schematic Workflow Depicting Combinatorial Enzymatic O-Glycoproteomics

Why Choose MtoZ Biolabs?

☑️Advanced Platforms: Equipped with Orbitrap and Q-TOF systems, our lab delivers high-resolution data with excellent sensitivity and confidence.

☑️Comprehensive Workflow: From enzymatic digestion to glycopeptide enrichment and data interpretation, all steps are optimized for precision.

☑️Customizable Solutions: Analytical strategies are tailored to the client's protein, research goal, and regulatory needs.

☑️Experienced Scientific Team: Our experts combine deep knowledge of glycoprotein chemistry and mass spectrometry to ensure reliable conclusions.

☑️Quality Assurance: Each project follows strict QC protocols, providing validated, publication-ready, and regulatory-compliant results.

FAQ

Q1: What Types of Samples Are Suitable?

A1: We accept a wide range of protein and biological samples, including purified proteins, recombinant proteins, antibodies, and other therapeutic proteins. Cell lysates, tissues, and biological fluids can also be submitted, with MtoZ Biolabs performing target protein extraction and preparation.

Q2: How Should I Prepare My Samples?

• All samples should be stored at –80°C.
• Use dry ice or ice packs during shipment to preserve protein integrity.
• Avoid freeze–thaw cycles that may alter disulfide linkages.

For more information, please refer to Sample Submission Guidelines for Proteomics.

Q3: What Is the Service General Workflow?

Q4: What Data Formats Are Provided?

Results are delivered in standard and easy-to-access formats, including:

• Raw MS data files (.raw, .wiff, .d)
• Processed data and quantification tables (.xlsx, .csv)
• Spectral and chromatogram images (.pdf, .png)
• Final analytical report (.pdf)

All deliverables are compatible with common data analysis software. Additional file formats or data structures can be provided upon request to meet specific research or publication requirements.

Start Your Project with MtoZ Biolabs

Contact us to discuss your experimental design or request a quote. MtoZ Biolabs' Targeted O-Glycosylation Site Mapping Service provides high-resolution, site-specific insights into the O-glycosylation patterns of target proteins.