A simple method for the release of asparagine-linked oligosaccharides from a glycoprotein purified by SDS-polyacrylamide gel electrophoresis

J Biochem. 1992 May;111(5):620-2. doi: 10.1093/oxfordjournals.jbchem.a123807.


A simple method for the release of oligosaccharides from glycoproteins separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) has been developed. Asialo-alpha 1-acid glycoprotein, which was tritiated at the nonreducing terminal D-galactopyranosyl residue by reduction with sodium borotritide after incubation with D-galactose oxidase, was used as a model compound. After electrophoretic separation of the glycoprotein, oligosaccharides were released by the use of a gas-phase hydrazinolysis apparatus. In the first method, the gel was stained with Coomassie Blue and the glycoprotein together with the gel was directly subjected to gas-phase hydrazinolysis after removal of water in a P2O5 desiccator. The recovery of released oligosaccharides was 25.9 +/- 2.4%, based on the amount of the glycoprotein loaded on the gel within the range of 3.5-28.5 micrograms. In the second method, the glycoprotein was electroblotted onto an Immobilon transfer membrane and was visualized by staining with Coomassie Blue. A small piece of the membrane with the corresponding band was cut out, dried in a desiccator and subjected to gas-phase hydrazinolysis. In this case, the recovery of released oligosaccharides was 15.2 +/- 1.0%. These procedures, particularly the first one, should be widely applicable for the isolation of oligosaccharides from glycoproteins separated by SDS-PAGE.

MeSH terms

  • Animals
  • Asialoglycoproteins / chemistry
  • Asialoglycoproteins / isolation & purification
  • Asparagine / chemistry
  • Carbohydrate Sequence
  • Electrophoresis, Polyacrylamide Gel
  • Glycoproteins / chemistry
  • Glycoproteins / isolation & purification*
  • Humans
  • Hydrazines / chemistry
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / isolation & purification
  • Molecular Sequence Data
  • Oligosaccharides / chemistry
  • Oligosaccharides / isolation & purification*
  • Orosomucoid / analogs & derivatives
  • Orosomucoid / chemistry
  • Orosomucoid / isolation & purification
  • Sodium Dodecyl Sulfate


  • Asialoglycoproteins
  • Glycoproteins
  • Hydrazines
  • Immunoglobulin G
  • Oligosaccharides
  • Orosomucoid
  • asialoorosomucoid
  • Sodium Dodecyl Sulfate
  • Asparagine