Glycan Sequencing by Tandem Mass Spectrometry
This experiment gives information on the type of oligosaccharides present, e.g high mannose, hybrid complex or mucin and also relative % of each oligosaccharide.
- We release, purify and derivate N– and O-linked glycans by enzymatic or chemical methods (PNGASE F, PNGASE A, Beta-elimination and hydrazinolysis) prior to MS Profiling. We can start from as small as 100μg of starting protein. The glycans can be prepared from a single purified glycoprotein or mixed protein and also from cells, tissues, culture supernatants, and gel slices.
- The ABISciex 5800 MALDI-TOF/TOF or the Thermo LTQ Orbitrap Fusion Tribrid mass spectrometers are then used for rapid glycan profiling, which is ideal for large sample sizes and projects.
- We can also provide N– and O– glycomics profiling by using labeling techniques with 2-aminobenzamide (2-AB), 2-aminobenzoic acid (2-AA), 2-aminopyridine (PA), 2-aminonaphthalene trisulfonic acid (ANTS), and 1-aminopyrene-3,6,8-trisulfonic acid (APTS) and their subsequent analysis by HPLC or LC-MS.