High-resolution mass spectrometry glycoprofiling of intact transferrin for diagnosis and subtype identification in the congenital disorders of glycosylation

Monique Van Scherpenzeel, Gerry Steenbergen, Eva Morava, Ron A. Wevers, Dirk J. Lefeber

Research output: Contribution to journalArticlepeer-review

55 Scopus citations


Diagnostic screening of the congenital disorders of glycosylation (CDG) generally involves isoelectric focusing of plasma transferrin, a robust method easily integrated in medical laboratories. Structural information is needed as the next step, as required for the challenging classification of Golgi glycosylation defects (CDG-II). Here, we present the use of high-resolution nano liquid chromatography-chip (C8)-quadrupole time of flight mass spectrometry (nanoLC-chip [C8]-QTOF MS) for protein-specific glycoprofiling of intact transferrin, which allows screening and direct diagnosis of a number of CDG-II defects. Transferrin was immunopurified from 10 μL of plasma and analyzed by nanoLC-chip-QTOF MS. Charge distribution raw data were deconvoluted by Mass Hunter software to reconstructed mass spectra. Plasma samples were processed from controls (n = 56), patients with known defects (n = 30), and patients with secondary (n = 6) or unsolved (n = 3) cause of abnormal glycosylation. This fast and robust method, established for CDG diagnostics, requires only 2 hours analysis time, including sample preparation and analysis. For CDG-I patients, the characteristic loss of complete N-glycans could be detected with high sensitivity. Known CDG-II defects (phosphoglucomutase 1 [PGM1-CDG], mannosyl (α-1,6-)-glycoprotein β-1,2-N-acetylglucosaminyltransferase [MGAT2-CDG], β-1,4-galactosyltransferase 1 [B4GALT1-CDG], CMP-sialic acid transporter [SLC35A1-CDG], UDP-galactose transporter [SLC35A2-CDG] and mannosyl-oligosaccharide 1,2-alpha-mannosidase [MAN1B1-CDG]) resulted in characteristic diagnostic profiles. Moreover, in the group of Golgi trafficking defects and unsolved CDG-II patients, distinct profiles were observed, which facilitate identification of the specific CDG subtype. The established QTOF method affords high sensitivity and resolution for the detection of complete glycan loss and structural assignment of truncated glycans in a single assay. The speed and robustness allow its clinical diagnostic application as a first step in the diagnostic procedure for CDG defects.

Original languageEnglish (US)
Pages (from-to)639-649.e1
JournalTranslational Research
Issue number6
StatePublished - Dec 1 2015


  • ATP6V0A2 ATPase, H+ transporting, lysosomal V0 subunit a2
  • Abbreviations ApoCIII apolipoprotein CIII
  • B4GALT1 β-1,4-galactosyltransferase 1
  • CDG Congenital Disorders of Glycosylation
  • CE Capillary electrophoresis
  • CMP cytidine monophosphate
  • COG1 component of oligomeric Golgi complex 1
  • CV Coefficient of variation
  • DPAGT1 dolichyl-phosphate (UDP-N-acetylglucosamine) N-acetylglucosaminephosphotransferase 1
  • EDTA Ethylenediaminetetraacetic acid
  • ESI Electron spray ionization
  • GDP guanosine diphosphate
  • HPLC High pressure liquid chromatography
  • HUS Hemolytic uremic syndrome
  • IEF Isoelectric focusing
  • LC Liquid Chromatography
  • MALDI Matrix assisted laser desorption ionization
  • MAN1B1 mannosyl-oligosaccharide 1,2-alpha-mannosidase
  • MGAT2 mannosyl(α-1,6-)-glycoprotein β-1,2-N-acetylglucosaminyltransferase
  • MS Mass spectrometry
  • NHS N-Hydroxysuccinimidyl
  • PGM1 Phosphoglucomutase 1
  • PMM2 phosphomannomutase 2
  • QTOF Quadrupole Time Of Flight
  • SLC35A1 CMP-sialic acid transporter
  • SLC35A2 UDP-galactose transporter
  • SLC35C1 GDP-fucose transporter
  • TIEF Transferrin Isoelectric focusing
  • TMEM165 transmembrane protein 165
  • Tf transferrin
  • Tris 2-Amino-2-(hydroxymethyl)-1,3-propanediol
  • UDP uridine disphosphate
  • VPS13B vacuolar protein sorting 13 homolog B

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health
  • Physiology (medical)
  • Biochemistry, medical


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