Permethylated N-glycan analysis with mass spectrometry

Methods Mol Biol. 2013:1007:289-300. doi: 10.1007/978-1-62703-392-3_12.

Abstract

Protein glycosylation plays an important role in multiple cell functions, and aberrations of protein glycosylation are associated with various malignancies including cancer. In this chapter, we provide a detailed protocol for MALDI MS analysis of permethylated N-glycans extracted from human serum proteins. The protocol includes procedures for N-glycan purification and in-solution permethylation, structural elucidation of permethylated N-glycans by MALDI-QIT-TOF MS, and construction of indices to quantify levels of certain types of glycosylation, such as fucosylation, which may serve as a potential disease biomarker.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biomarkers, Tumor / analysis
  • Blood Proteins / chemistry*
  • Glycated Serum Proteins
  • Glycoproteins / chemistry*
  • Humans
  • Mass Spectrometry / methods*
  • Neoplasms / metabolism
  • Polysaccharides / analysis*
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Biomarkers, Tumor
  • Blood Proteins
  • Glycoproteins
  • Polysaccharides
  • Glycated Serum Proteins