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Nanoproteomics

Volume 790 of the series Methods in Molecular Biology pp 87-97

Date:

N-Linked Global Glycan Profiling by NanoLC Mass Spectrometry

  • Michael S. BeremanAffiliated withDepartment of Genome Sciences, University of Washington
  • , David C. MuddimanAffiliated withDepartment of Chemistry, North Carolina State University Email author 

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Abstract

A method is detailed for the global profiling of underivatized N-linked glycans that are derived from complex protein mixtures. The method consists of five main steps that include the following: (1) protein denaturation; (2) enzymatic digestion; (3) solid phase extraction; (4) nanoLC MS analysis; and (5) data interpretation. Materials, methods, and algorithms for the identification of both glycan composition and structure are summarized. In addition, potential problems and their resolutions are addressed.

Key words

N-linked glycans NanoLC mass spectrometry Glycoproteins Glycosylation Glycan profiling Hydrophilic interaction chromatography Glycobiology Glycan algorithms