The biology of carbohydrates, or glycobiology, is attracting growing attention in high impact research areas (neurobiology, development, immunology), and also in applied fields such as manufacturing of biotherapeutics and antibodies, and heparin analogs. Carbohydrates attached to glycolipids or glycoproteins determine the properties and fates of living cells, therefore the structural characterization of glycans and glycosylated proteins plays a crucial role in elucidating central biological processes.
Glycoproteins and other glycoconjugates are structurally diverse and virtually impossible to predict from genetic information alone. Analytical tools, including specific enzymes such as endoglycosidases and exoglycosidases, along with chromatography, mass spectrometry methods and specialized bioinformatics, allow us to define the composition of glycoconjugates in any given model system. However, descriptive analysis only gives a snapshot of the biological process of interest. In many cases, purifying a target from a recombinant expression system is the preferred method to understand the functional properties of a glycoprotein.
Glycobiology & Proteomics includes these areas of focus:
FAQs for Glycobiology & Proteomics
- How can I separate a deglycosylated protein from the glycosidase?
- What are the best techniques to analyze N- and O-glycans?
- What are the advantages of enzymatic deglycosylation?
- Can glycosidases be used in combination for extensive digestion?
- Do I need to deglycosylate my protein sample for proteomic analysis?
- What are glycosidases and their uses?
- Can I analyze protein glycosylation with only benchtop instrumentation and reagents?
- Is it possible to predict whether a protein is N- or O-glycosylated?
- Which proteins are glycosylated?
- Detailed characterization of antibody glycan structure using exoglycosidases
- Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis
The Glycoproteomics brochure provides information on the suite of endo- and exoglycosidases, and deglycosylation enzymes offered by NEB.
Glycoproteomics Technical Guide
Find in-depth information, including protocols, technical tips, frequently asked questions and application notes, utilizing NEB’s suite of endo- and exoglycosidases.
Rapid PNGase F Trifold
Learn how Rapid PNGase F can speed up your antibody deglycosylation step and make your therapeutic proteins faster and more efficiently than ever before.
Glycomics: A rapidly evolving field with a sweet future
Glycans are widely distributed in nature and have physical, chemical, and biological properties that make them important players in areas such as biofuels, food, materials science, biotechnology, and pharmaceuticals.
N-Glycan Composition Profiling for Quality Testing of Biotherapeutics
N-Glycan Composition Profiling for Quality Testing of Biotherapeutics
The Structure, Function and Importance of Carbohydrates
Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
- Endoglycosidase Selection Chart
- Detailed Characterization of Several Glycosidase Enzymes
- Glycobiology Unit Conversion Chart
Other Tools & Resources
- Stech M, Quast RB, Sachse R, Schulze C, Wüstenhagen DA, Kubick S 2014. A continuous-exchange cell-free protein synthesis system based on extracts from cultured insect cells PLoS One. 9(5), PubMedID: 24804975, DOI: 10.1371/journal.pone.0096635
- Kwon HM, Lee KH, Han BW, Han MR, Kim DH, Kim DE 2014. An RNA aptamer that specifically binds to the glycosylated hemagglutinin of avian influenza virus and suppresses viral infection in cells PLoS One. 9(5), PubMedID: 24835440, DOI: 10.1371/journal.pone.0097574
- Botto L, Cunati D, Coco S, Sesana S, Bulbarelli A, Biasini E, Colombo L, Negro A, Chiesa R, Masserini M, Palestini P 2014. Role of lipid rafts and GM1 in the segregation and processing of prion protein PLoS One. 9(5), PubMedID: 24859148, DOI: 10.1371/journal.pone.0098344
- Rosenbaum EE, Vasiljevic E, Brehm KS, Colley NJ 2014. Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster PLoS Genet. 10(5), PubMedID: 24785692, DOI: 10.1371/journal.pgen.1004349
- Rosenbaek LL, Kortenoeven ML, Aroankins TS, Fenton RA 2014. Phosphorylation decreases ubiquitylation of the thiazide-sensitive cotransporter NCC and subsequent clathrin-mediated endocytosis J Biol Chem. 289(19), PubMedID: 24668812, DOI: 10.1074/jbc.M113.543710
- Möykkynen T, Coleman SK, Semenov A, Keinänen K 2014. The N-terminal domain modulates α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization J Biol Chem. 289(19), PubMedID: 24652293, DOI: 10.1074/jbc.M113.526301
- Arakel EC, Brandenburg S, Uchida K, Zhang H, Lin YW, Kohl T, Schrul B, Sulkin MS, Efimov IR, Nichols CG, Lehnart SE, Schwappach B 2014. Tuning the electrical properties of the heart by differential trafficking of KATP ion channel complexes J Cell Sci. 127(Pt 9), PubMedID: 24569881, DOI: 10.1242/jcs.141440
- Itahana Y, Han R, Barbier S, Lei Z, Rozen S, Itahana K 2014. The uric acid transporter SLC2A9 is a direct target gene of the tumor suppressor p53 contributing to antioxidant defense Oncogene. , PubMedID: 24858040, DOI: 10.1038/onc.2014.119
- Wicht O, Burkard C, de Haan CA, van Kuppeveld FJ, Rottier PJ, Bosch BJ 2014. Identification and Characterization of a Proteolytically Primed Form of the Murine Coronavirus Spike Proteins after Fusion with the Target Cell J Virol. 88(9), PubMedID: 24554652, DOI: 10.1128/JVI.03451-13
- Wright CR, Brown EL, Della-Gatta PA, Ward AC, Lynch GS, Russell AP 2014. G-CSF does not influence C2C12 myogenesis despite receptor expression in healthy and dystrophic skeletal muscle Front Physiol. 5, PubMedID: 24822049, DOI: 10.3389/fphys.2014.00170
- Zhao H, Blazanovic K, Choi Y, Bailey-Kellogg C, Griswold KE 2014. Gene and protein sequence optimization for high-level production of fully active and aglycosylated lysostaphin in Pichia pastoris Appl Environ Microbiol. 80(9), PubMedID: 24561590, DOI: 10.1128/AEM.03914-13
- Haller G, Li P, Esch C, Hsu S, Goate AM, Steinbach JH 2014. Functional characterization improves associations between rare non-synonymous variants in CHRNB4 and smoking behavior PLoS One. 9(5), PubMedID: 24804708, DOI: 10.1371/journal.pone.0096753
- Velho, A.M., Jarvis, S.M. 2009. Topological studies of hSVCT1, the human sodium-dependent vitamin C transporter and the influence of N-glycosylation on its intracellular targeting Exp. Cell Res. . 315 , PubMedID: 19379732, DOI:
- Boeggeman, E., Ramakrishnan, B., Pasek, M., Manzoni, M., Puri, A., Loomis, K.H., Waybright, T.J., Qasba, P.K. 2009. Site specific conjugation of fluoroprobes to the remodeled Fc N-glycans of monoclonal antibodies using mutant glycosyltransferases: application for cell surface antigen detection Bioconjug. Chem. . 20 , PubMedID: 19425533, DOI:
- Gong, B., Cukan, M., Fisher, R., Li, H., Stadheim, T.A., Gerngross, T. 2009. Characterization of N-linked glycosylation on recombinant glycoproteins produced in Pichia pastoris using ESI-MS and MALDI-TOF Methods Mol. Biol. . 534 , PubMedID: , DOI:
- Rasmussen, T.N., Plenge, P., Bay, T., Egebjerg, J., Gether, U. 2009. A single nucleotide polymorphism in the human serotonin transporter introduces a new site for N-linked glycosylation Neuropharmacology . 57 , PubMedID: 19500602, DOI:
- Gefter, J.V., Shaufl, A.L., Fink, M.P., Delude, R.L. 2009. Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines Cell Tissue Res. . 337 , PubMedID: 19415334, DOI:
- Wagner-Rousset, E., Bednarczyk, A., Bussat, M.C., Colas, O., Corvaïa, N., Schaeffer, C., Van Dorsselaer, A., Beck, A. 2008. The way forward, enhanced characterization of therapeutic antibody glycosylation: comparison of three level mass spectrometry-based strategies J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.. 872, PubMedID: 18672411, DOI:
- Graham, D.R., Mitsak, M.J., Elliott, S.T., Chen, D., Whelan, S.A., Hart, G.W., Van Eyk, J.E. 2008. Two-dimensional gel-based approaches for the assessment of N-Linked and O-GlcNAc glycosylation in human and simian immunodeficiency viruses Proteomics. 872, PubMedID: 19072736, DOI:
- Wong-Madden, S.T., Landry, D., and Guthrie, E.P. 1997. Discovery and Uses of Novel Glycosidases Techniques in Glycobiology. , PubMedID: , DOI:
Publications related to Glycobiology & Proteomics
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Learn about glycobiology and its importance in clinical and diagnostic applications in this episode of NEB TV. Also, hear more about how NEB is setting the bar for product quality in this rapidly growing field.
Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans can be accomplished with the use of deglycosylation enzymes, which can provide complete sugar removal with no protein degradation.