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Glycoproteins participate in various biological processes, but glycoproteins analysis is a big challenge because of the low abundance and heterogeneity of glycans. As a forward-looking company as well as a leading customer service provider, Creative Biolabs has successfully explored a series of innovative glycomic profiling platforms to help our clients identify an entire set of N- and O-glycans with high sensitivity and specificity in the biological material. We are confident to help you get milestone success in your glycoprotein project.
Glycosylation a fundamental post-translational modification, which plays an important role in many physiological and pathological processes. Glycoprotein structure analysis allows characterization of important biological molecules and would be of great diagnostic and clinical importance. As an important part of glycoprotein structure analysis, glycomic profiling has attracted a lot of attention.
Glycomic profiling can identify an entire set of N-, and O-glycans present in a diverse range of biological material, including secretions, cell lines, tissues, and organs, which refers to the collection, analysis, and exploitation of glyco-biomaterial at the glycome level. Profiling of glycomic changes in a cell or organism level can be used to provide a general overview on the glycome, the total glycosylation pattern of glycoproteins, glycolipids, or other types of biomolecules, which is increasingly important in searching biomarkers for diseases. N-glycomic and O-glycomic profiling are two common types of analysis. These glycomic strategies are based on rapid, high-sensitivity mass spectrometric (MS) strategies, combined with electrospray (ES) or MALDI tandem mass spectrometry (MS/MS) sequencing and gas chromatography (GC)-MS linkage analysis.
N-glycosylation usually occurs on the asparagine residue of proteins that shuttle via the secretory pathway. As a good complement to genomic and proteomic studies, N-glycomic profiling is widely used in the analysis of the entire repertoire of N-glycans of an organism such as glycans attached to proteins and lipids, glycosaminoglycans and polysaccharides, including genetic, physiologic, pathologic, and other aspects. Moreover, N-glycomic profiling has the diagnostic potential for different stages of cancers and various other disease states, which could provide new biomarkers. Technologies for N-Glycomic profiling in Creative Biolabs including but not limited to:
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a high-throughput method to analyze the structural characterization of glycosylated compounds with high sensitivity and robustness. MALDI-TOF MS can identify the entire set of N-glycans expressed by plasma/serum, cell, tissue or organism. During the detection, all glycans attached to the proteins will release by enzymatic digestion, and then separated by hydrophilic chromatography and finally quantitatively profiled with MALDI-TOF MS system.
Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) is a useful tool to analyze N-Glycomic. This protocol permits the detection of permethylated N-glycans in large tissue samples.
Fig.1 N-glycomic profiling of tissue sections by LC-MS (Hu, 2013).
O-glycosylations includes two common types, namely O-acetylgalactosamine (O-GalNAc) and O-acetylglucosamine (O-GlcNAc), among which, O-GalNAc is usually attached to the hydroxyl group of the protein serine or threonine residues through an α-linkage, while O-GlcNAc is attached through a β-linkage. Unlike N-glycans, there is no known O-linked amino acid consensus sequence yet. O-Glycomic profiling is an important part of genomic and proteomic studies.
Technologies for O-Glycomic profiling in Creative Biolabs including but not limited to:
Workflow of Glycomic Profiling
Fig.2 Glycomics workflow by MALDI-TOF MS (Jansen, 2016).
Creative Biolabs has an experienced team of dedicated scientists offering a wide range of glycomic analysis. Our team has successfully completed a lot of projects for glycomic and proteomic analysis. We will provide reliable results to drive your scientific research. We can also customize our offering to meet your specific project needs. If you are interested, please contact us for more information.