O-Glycosylation Site Analysis Service

O-glycosylation is another important protein glycosylation modification besides N-glycosylation. Unlike N-glycosylation, O-glycan chains are usually attached to serine (Ser) or threonine (Thr) residues. O-glycosylation site analysis typically involves multiple steps, including protein extraction, proteolytic digestion, enrichment of O-glycosylated peptides, and liquid chromatography-mass spectrometry (LC-MS) analysis. By analyzing O-glycosylation sites, detailed information about the protein's O-glycosylation status can be obtained, and how this post-translational modification affects the protein's function and activity, thus, it has also been applied in the research and development of new drugs and the production and optimization of protein drugs.

 

MtoZ Biolabs based on the principles of glycosylation site analysis, has established a stable and efficient glycosylation site analysis platform using high-performance liquid chromatography combined with high-resolution mass spectrometry (HPLC-MS/MS), providing O-glycosylation site analysis services for proteins/peptides, antibodies, protein vaccines, recombinant collagen, and other bioproducts, including O-glycan analysis, O-glycosylation site identification, and a complete analysis process of glycopeptides. Free project evaluation!

 

Experimental Equipment

1. NanoLC: Vanquish NEO 

2. MS: Orbitrap Fusion Lumos With ETD

 

Case Study

We use a direct analysis method of O-glycopeptides for O-glycosylation site analysis. Firstly, an appropriate protease is selected based on the protein's theoretical amino acid sequence information to cleave the protein into peptides. Then, mass spectrometry raw data are collected using HPLC-MS/MS. This data is imported into an O-glycan database, and O-glycosylation modifications are added as variable modifications to the data analysis. Finally, it determines which sites in the protein have undergone O-glycosylation modification.

 



Figure 2. O-Glycosylation Site Secondary Diagram

Note: In the diagram, the O-glycosylation modification occurs at the fifth amino acid of the peptide, the molecular weight change is +365.132, the horizontal axis is the mass-to-charge ratio, the vertical axis is signal strength.

 

Deliverables

In the technical report, MtoZ Biolabs will provide you with detailed technical information, including:

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. O-Glycosylation Site Analysis Details

4. Mass Spectrometry Images

5. Raw Data