N-linked glycan profiling of mature human milk by high-performance microfluidic chip liquid chromatography time-of-flight tandem mass spectrometry

David C. Dallas, William F. Martin, John S. Strum, Angela M. Zivkovic, Jennifer T. Smilowitz, Mark Underwood, Michael Affolter, Carlito B Lebrilla, J. Bruce German

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

N-Linked glycans of skim human milk proteins were determined for three mothers. N-Linked glycans are linked to immune defense, cell growth, and cell-cell adhesion, but their functions in human milk are undetermined. Protein-bound N-linked glycans were released with peptidyl N-glycosidase F (PNGase F), enriched by graphitized carbon chromatography, and analyzed with Chip-TOF MS. To be defined as N-glycans, compounds were required, in all three procedural replicates, to match, within 6 ppm, against a theoretical human N-glycan library and be at least 2-fold higher in abundance in PNGase F-treated than in control samples. Fifty-two N-linked glycan compositions were identified, and 24 were confirmed via tandem mass spectra analysis. Twenty-seven compositions have been found previously in human milk, and 25 are novel compositions. By abundance, 84% of N-glycans were fucosylated and 47% were sialylated. The majority (70%) of total N-glycan abundance was composed of N-glycans found in all three milk samples.

Original languageEnglish (US)
Pages (from-to)4255-4263
Number of pages9
JournalJournal of Agricultural and Food Chemistry
Volume59
Issue number8
DOIs
StatePublished - Apr 27 2011

Keywords

  • glycobiology
  • glycomics
  • glycoprotein
  • glycoproteomics
  • human milk
  • LC-MS
  • N-linked glycan
  • oligosaccharide
  • prebiotics
  • sialic acid
  • time-of-flight mass spectrometry

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Chemistry(all)

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