Removal of N-Linked & O-Linked Glycans from Glycoproteins

Removal of oligosaccharides from glycoproteins, termed deglycosylation, is often used in order to simplify analysis of the peptide and/or glycan portion of a glycoprotein. Detailed knowledge of the glycan structures helps to correlate them to their respective function. To do this, tools are required for highly sensitive analysis of glycan chains. Both chemical and enzymatic methods exist for removing oligosaccharides from glycoproteins. However, chemical methods such as β-elimination with mild alkali (1) or mild hydrazinolysis (2) often results in the degradation of the protein; whereas, enzymatic methods such as treating with PNGase F (NEB #P0704, NEB #P0705) or Endo H (NEB #P0702, NEB #P0703) (3) for N-glycans are much gentler and can provide complete sugar removal with no protein degradation. Unfortunately, there are no general endoglycosidases that can give complete removal of all O-linked glycans. The broadest specificity O-glycosidase (NEB #P0733, NEB #E0540) is able to cleave only core 1 and core 3 O-glycans (4). Using the O-glycosidase in combination with an appropriate mix of exoglycosidases allows the removal of large O-glycans, after the exoglycosidases have trimmed them to their core. However, for removal of intact O-glycan chains a researcher must still use chemical methods.

Using Glycosidases to Remove, Trim, or Modify Glycans on Therapeutic


  1. Kakehi K., et al (1994) J Chromatogr A 680, 209–215. PMID: 7952002
  2. Royle L., et al (2002) Anal Biochem 304, 70–90. PMID: 11969191
  3. Maley, F. et al. (1989) Anal. Biochem., 180, 195-204. PMID: 2510544
  4. Koutsioulis, D. et al (2008) Glycobiology , 18, 799-805. PMID: 18635885
  1. Identification and Characterization of Protein Glycosylation P6039S

    Learn how glycosidases are used to analyze multiple glycoprotein samples. Here, the model glycoprotein, hCGβ, which carries both N- and O-glycans, is demonstrated.

  2. Overview of Glycobiology

    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.

  3. Elucidating the Complexity of Heparin Oligosaccharide Analysis

    Learn more about the structural elucidation of complex and diverse heparin oligosaccharides using Bacteroides Heparinase I, II and III in combination with downstream mass spec analysis.

FAQs for Removal of N-Linked & O-Linked Glycans from Glycoproteins

Protocols for Removal of N-Linked & O-Linked Glycans from Glycoproteins

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