N-glycosylation is a post-translational modification of proteins involved in various physiological and pathological processes. The variability of glycosylation modification sites is a key indicator of cellular activity, as evidenced by the correlation between the reduction of glycosylation attachment sites in serum proteins and the severity of congenital disorders of glycosylation (CDGs). Therefore, accurately determining site occupancy is significant for fully understanding the impact of protein glycosylation on human health.
To determine glycosylation attachment sites, PNGase F is typically used in heavy water to separate polysaccharides from proteins. During this process, the previously glycosylated asparagine undergoes deamidation to become aspartic acid, with an increase in O18 modification. Due to the molecular mass change after the introduction of O18, glycosylation sites can be detected by comparing peptides containing aspartic acid with those containing non-glycosylated asparagine. The general strategy for analyzing variable N-glycosylation sites in glycoproteins includes protein digestion with PNGase F enzyme, glycan release, liquid chromatography separation of glycopeptides, and mass spectrometry detection.
General analysis workflow for LC-MS-based N-glycosylation site attachment analysis
BiotechPack has extensive experience in the field of glycomics analysis services and can tailor glycomics analysis services based on your specific needs. Welcome to consult us!
Project report in Chinese/English
In the technical report, BiotechPack will provide you with a detailed bilingual technical report in both Chinese and English, including:
1. Experimental procedures (Chinese and English)
2. Relevant mass spectrometry parameters (Chinese and English)
3. Mass spectrometry images
4. Raw data
5. Detailed information on identified glycosylation sites
One-stop service for N-glycosylation site analysis
You only need to place an order and send your samples
BiotechPack completes the one-stop service: sample processing - on-machine analysis - data analysis - project report
