O-Glycan Analysis Service

O-glycosylation, a complex and diverse modification within the vast realm of protein modifications, substantially enriches protein functionality. Unlike N-glycosylation, O-glycosylation poses significant challenges due to the complex nature of its glycan structures and the difficulty in accurate characterization. This modification predominantly involves attaching O-linked glycan chains to Serine, Threonine, or occasionally Tyrosine residues of proteins, with O-linked N-acetylgalactosamine (O-GalNAc) being the most prevalent.

 



Madunić, K. et al. Cell Mol Life Sci. 2021.

Figure1. O-Glycan Profiles From Two Exemplary CRC Cell Lines

 

Services at MtoZ Biolabs

MtoZ Biolabs offers a comprehensive O-glycan analysis service from sample preparation to in-depth analysis, employing advanced mass spectrometry techniques such as Matrix-Assisted Laser Desorption/Ionization (MALDI) MS and Electrospray Ionization (ESI) MS for superior data quality.

 

Analysis Workflow

1. Protein Denaturation

Modify the protein’s three-dimensional structure via chemical or physical means to expose the O-glycosylation sites.

2. N-glycan Removal

Employ PNGase F to eliminate any N-glycans that may interfere with the analysis.

3. O-glycan Release

Utilize reductive alkaline β-elimination to liberate O-glycans.

4. Glycan Enrichment and Separation

Apply Hydrophilic Interaction Liquid Chromatography (HILIC) to enrich the released O-glycans.

5. Glycan Labeling

Convert all hydroxyl groups to methyl ethers through permethylation and stabilize carboxyl groups through methylesterification for enhanced HPLC and MS analysis.

6. Mass Spectrometry Analysis

Conduct detailed analyses using MALDI-TOF MS or ESI-MS.

7. Report Generation

Deliver a detailed report encompassing experimental procedures, chromatographic and mass spectrometric parameters, raw data, and bioinformatics analysis.

 

Service Advantages

1. High analytical sensitivity and specificity ensure dependable O-glycan characterization.

2. Suitable for diverse biological samples, including complex cell and tissue matrices.

3. Guarantees extensive coverage and swift turnaround for O-glycan analysis.

4. Offers tailor-made, comprehensive services from experimental design to data interpretation.

5. Extensive bioinformatics analyses elucidate the biological implications of O-glycans.

 

In accordance with the rigorous ICH Q6B guidelines, MtoZ Biolabs leverages its skilled technical team and extensive experimental expertise to provide thorough O-glycan analysis services. For inquiries or special requests, please feel free to contact us. We eagerly anticipate collaborating with you.