Comparison of fluorescent labels for oligosaccharides and introduction of a new postlabeling purification method

Martin Pabst, Daniel Kolarich, Gerald Pöltl, Thomas Dalik, Gert Lubec, Andreas Hofinger, Friedrich Altmann

Research output: Contribution to journalArticleScientificpeer-review

111 Citations (Scopus)

Abstract

Labeling of oligosaccharides with fluorescent dyes is the prerequisite for their sensitive analysis by high-performance liquid chromatography (HPLC). In this work, we present a fast new postlabeling cleanup procedure that requires no device other than the reaction vial itself. The procedure can be applied to essentially all labeling reagents. We also compare the performance of 15 different labels for N-glycan analysis in various analytical procedures. We took special care to prevent obscuring influences from incomplete derivatization and signal quenching by impurities. Procainamide emerged as more sensitive than anthranilic acid for normal-phase HPLC, but its chromatographic performance was not convincing. 2-Aminopyridine was the label with the lowest retention on reversed-phase and graphitic carbon columns and, thus, appears to be most suitable for glycan fractionation by multidimensional HPLC. Most glycan derivatives performed better than native sugars in matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and electrospray ionization-MS (ESI-MS), but the gain was small and hardly sufficient to compensate for sample loss during preparation.

Original languageEnglish
Pages (from-to)263-273
Number of pages11
JournalAnalytical Biochemistry
Volume384
Issue number2
DOIs
Publication statusPublished - 15 Jan 2009

Keywords

  • Fluorescent label
  • Glycoprotein
  • N-Glycan

Fingerprint

Dive into the research topics of 'Comparison of fluorescent labels for oligosaccharides and introduction of a new postlabeling purification method'. Together they form a unique fingerprint.

Cite this