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.
- What are glycosidases and their uses?
- Which proteins are glycosylated?
- Can glycosidases be used in combination for extensive digestion?
- Can I analyze protein glycosylation with only benchtop instrumentation and reagents?
- Do I need to deglycosylate my protein sample for proteomic analysis?
- How can I separate a deglycosylated protein from the glycosidase?
- Is it possible to predict whether a protein is N- or O-glycosylated?
- What are the advantages of enzymatic deglycosylation?
- What are the best techniques to analyze N- and O-glycans?
The Structure, Function and Importance of Carbohydrates
Read about the structure, function, and importance of Carbohydrates from biology experts at NEB.
Using Glycosidases to Remove Trim or Modify Glycans on Therapeutic Protein
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
O-glycan analysis of therapeutic proteins enabled by O-glycoprotease
This article from European Pharmaceutical Review describes tools that can be used for O-glycan analysis of therapeutic proteins.
- Glycoproteomics Brochure
- Glycoproteomics Technical Guide
- Rapid PNGase F Trifold
- Glycan Analyzer
- Endoglycosidase Selection Chart
- Detailed Characterization of Several Glycosidase Enzymes
- Glycobiology Unit Conversion Chart
- Improving Sample Workflow: Rapid PNGase F for Accurate Antibody Characterization (2015)
- Chuzel, L.O., Ganatra, M.B., Rapp, E., Henrissat, B., Taron, C.H. (2018) Functional metagenomics identifies an exosialidase with an inverting catalytic mechanism that defines a new glycoside hydrolase family (GH156) J Biol Chem; 293(47), 18138-18150. PubMedID: 30249617
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- 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), e97574. PubMedID: 24835440, DOI: 10.1371/journal.pone.0097574
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- 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), 4943-52. PubMedID: 24554652, DOI: 10.1128/JVI.03451-13
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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.