Glycomics and Glycoproteomics

Glycomics and Glycoproteomics The cell surface of all living organisms is covered with complex carbohydrates (Glycans) of different types. Glycomics is the comprehensive study of the Glycome. This includes the entire repertoire of sugars of an organism such as glycans attached to proteins and lipids, glycosaminoglycans and polysaccharides, including genetic, physiologic, pathologic, and other aspects. The rapid advancement of technologies such as high-throughput mass spectrometry, glycan microarrays and carbohydrate chemistry are helping to unravel the complexity resulting from these diverse molecules and eventually the ‘glycan code’. Mission: To establish and to cater comprehensive structural characterization of repertoire of complex carbohydrates present in proteins, cells, tissues and an organism. This is the first laboratory to provide niche technology service in India and aims to be one of the best in the world. The diversity nature of these molecules include glycans attached to proteins (N-, O-glycans and O-GlcNAcylatoin), glycans bound to lipids (glycolipids), glycosaminoglycans, other oligo and polysaccharides. Values: We hope to achieve our mission through; i) developing specific standard protocols using state of the art technologies and instrumentation for each of the different categories of these diverse molecules, ii) strong commitment to quality of the data produced, iii) reach-out to scientists in academia and industry to familiarize about our capabilities and help in their scientific endeavor, iv) developing pool of experts in carbohydrate analysis by quality training, and v) to be on the cutting-edge and meet future challenges by constantly improving and developing novel technologies.	SUBMIT A REQUEST
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OUR CAPABILITIES

Through our recently established Glycomics and Glycoproteomics Facility at C-CAMP we provide targeted services to help better understand the nature of glycans and glycoproteins and the impact of sugar modifications on proteins.

Services

The following services are being offered through our Glycomics and Glycoproteomics facility:

1. Glycomics of proteins, cells, tissue and whole organism

Structural characterization of N- and O-glycans attached to proteins. This will be performed using ultra sensitive MALDI-TOF/TOF MS in conjunction with chemical and enzymatic sample preparation techniques. The linkage between different sugar units will be ascertained using GC-MS, exoglycosidases and glycosyltransferases depending upon the requirement.N-glycan profiling of 2-AB derivatized glycans using UHPLC with fluorescent detector is also established.

2. Glycoproteomics

Identification of nature the glycans and their attachment site in a glycoprotein using a combination of conventional glycan analysis (MALDI-TOF/TOF) and glycopeptide analysis by NanoLC-ESI-MS/MS and offline-LC-MALDI-TOF/TOF.

3. Monosaccharide Composition Analysis

This analysis will provide information about relative intensities of various saccharides (sugars) of glycans, polysaccharides, etc. This study will be performed using acetylation and/or trimethylsilyl derivatization followed by GC-MS analysis

4. Glycosaminoglycans (coming soon)

Glycosaminoglycans are acidic, sulfated linear polysaccharides with disaccharide repeats including heparan sulfate, chondroitin sulfates and hyaluronic acids. These molecules are indicated in development, signaling, pattern formation, neuronal path-finding, axon guidance etc., and disease.Establishment of methods to characterize the structure of GAGs and their quantification will spur growth in the field of glycobiology. We intend to use a combination of enzyme digestion and HPLC, LC-MS/MS and capillary electrophoresis with LIF detection for the structural analysis.

5. Glycans attached to lipids

Glycolipids play a number of important functions in physiology and recognition by pathogens.Glycolipids will be isolated from cells, tissue and organism by standard protocol and the glycans will be released enzymatically using glycanases. The released glycans will be permethylated and analyzed by MALDI-TOF/TOF. Alternatively, whole glycolipid can be subjected to MALDI-TOF/TOF analysis to obtain information about the lipids and bound glycans.

6. Polysaccharide analysis

A large variety of carbohydrates other than those described above could come under polysaccharides including lipopolysaccharides (LPS), polysaccharides from plants used for bio-fuel industry etc. We have well established so methods for the structural characterization of polysaccharides using HPLC and mass spectrometry based techniques.

Biologics Characterization Facility

C-CAMP has established an analytical laboratory for characterization of structure and physicochemical properties of a biologic (protein) or biosimilar. A dedicated Nano-LC/MS/MS system along with size exclusion chromatography, capillary isoelectric focusing, CD, fluorescence, analytical ultra centrifuge, DSC etc will be used to characterize the biologics/biosimilars using validated analytical (GLP/NABL) protocols and instrumentation

TECHNOLOGY PLATFORMS

UPLC with diode array and flourescence detectors