Glycomic Profiling
Glycomic Profiling of N-glycans and O-glycans by Mass Spectrometry (MS)
Glycomic analysis by Mass Spectrometry
Mass spectrometry profiling of glycans released from purified glycoproteins, glycolipids, as well as cultured cells, tissues, or organs can generate an overall picture of the glycome, including monosaccharide compositions of glycans and certain level of structural prediction. This can be carried out by EGMIC staffs using different types of mass spectrometers. In addition, EGMIC provides access to instrument at an hourly rate for your glycomic or glycoproteomic analyses.
We provide services in preparation of glycolipids, N-glycans and O-glycans from glycoproteins, serum, cells or tissue. Products can be used for glycomics profiling by Mass Spectrometry (MS) or further used for shotgun glycan microarray development.
Detailed analysis of glycans
Compositional analysis and predicted structural analyses are informative, but in many cases actually detailed structures analyses are required to support your project. In addition to Agilent IM-QTOF-MS hosted by EGMIC, we have access to other instruments through the Emory Integrated Proteomics Core. Also, since Emory and UGA have agreed on reciprocal use of core facilities, our core will be able to work with Emory investigators to access resources at the University of Georgia Complex Carbohydrate Research Center (CCRC) in Athens to support these more detailed analyses. Cost for these services will be based on the CCRC cost structure and determined based on nature of the request.
For a serum sample, 25 µl are needed for N-glycan profiling and 25 µl are needed for O-glycan profiling. The starting material for cell sample is 2-10 E10 cells for mass profiling. More cells are needed to prepare the glycans for a shotgun glycan microarray development.
For purified glycoprotein, 50-100 µg are needed. If your sample is a mixture containing glycoprotein, 1 – 2 mg is needed depending on glycoprotein content. If any additive or ingredient in your sample mayinterfere with the process, dialysis will be performed to achieve a cleaner sample.
We normally use 100ug Ribonuclease B, human transferrin or fetuin as process controls for N-glycans preparation and 100ug fetuin or mucin as process control for O-glycan release. See Methods section for general protocol.