N-Glycan Profiling and Its Analysis Workflow

N-glycan analysis is a method used to analyze glycan structures, especially when studying glycosylation on proteins or other biomolecules. Glycosylation (or glycation) is a common mode of biomolecular modification in cells, playing a key role in protein stability, activity, cell signaling, cell-cell interaction, and more.

 

N-glycan refers to the glycan chain attached to the asparagine nitrogen, which is a type of N-linked glycoprotein. N-linked glycoproteins are widely present in organisms and are crucial for many physiological processes.

 

Analysis Workflow

1. Sample Preparation

First, the target protein or peptide needs to be extracted.

 

2. Glycosidase Digestion

A specific glycosidase (such as PNGase F) is used to release the N-linked glycans from the protein or peptide.

 

3. Glycan Purification

After glycosidase digestion, the glycans need to be purified from the mixture.

 

4. Labeling Derivatization

To improve the sensitivity of the analysis or for better separation, the glycans can be labelled or derivatized.

 

5. Analysis

Chromatography, mass spectrometry or other techniques are used to qualitatively and quantitatively analyze the glycans.

 

6. Data Interpretation

Based on the obtained data, the structure and composition of the glycans are interpreted.