High-Throughput Automated Micro-permethylation for Glycan Structure Analysis

Anal Chem. 2019 Jan 15;91(2):1237-1240. doi: 10.1021/acs.analchem.8b05146. Epub 2018 Dec 21.

Abstract

Permethylation is an essential technique for the detailed structural characterization of glycans by mass spectrometry. However, it requires specialized skills and equipment and is labor-intensive. While this limits glycan analysis to a handful of experts, the increasing awareness of the tremendous importance of glycans in physiological processes of health and disease has drastically raised the demand for detailed structural characterization of glycans. We have developed a simple microplate permethylation method that relies upon solid phase extraction using C18 tips to purify the permethylated glycans. The procedure is easy to perform, making it accessible to nonexperts, and fast, promising to accelerate glycan research. A 96-well plate of released glycan samples can be permethylated in less than an hour. The procedure can be carried out without discontinuous steps in an automatic pipet and integrated seamlessly with glycan release and mass spectrometry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Methylation
  • Molecular Structure
  • Polysaccharides / chemistry*
  • Polysaccharides / isolation & purification
  • Solid Phase Extraction / methods*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Polysaccharides