Stable isotopic labeling-based quantitative targeted glycomics (i-QTaG)

Kyoung Jin Kim, Yoon Woo Kim, Yun Gon Kim, Hae Min Park, Jang Mi Jin, Young Hwan Kim, Yung Hun Yang, Jun Kyu Lee, Junho Chung, Sun Gu Lee, Alan Saghatelian

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Mass spectrometry (MS) analysis combined with stable isotopic labeling is a promising method for the relative quantification of aberrant glycosylation in diseases and disorders. We developed a stable isotopic labeling-based quantitative targeted glycomics (i-QTaG) technique for the comparative and quantitative analysis of total N-glycans using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We established the analytical procedure with the chemical derivatizations (i.e., sialic acid neutralization and stable isotopic labeling) of N-glycans using a model glycoprotein (bovine fetuin). Moreover, the i-QTaG using MALDI-TOF MS was evaluated with various molar ratios (1:1, 1:2, 1:5) of 13C6/12C6-2-aminobenzoic acid-labeled glycans from normal human serum. Finally, this method was applied to direct comparison of the total N-glycan profiles between normal human sera (n=8) and prostate cancer patient sera (n=17). The intensities of the N-glycan peaks from i-QTaG method showed a good linearity (R2>0.99) with the amount of the bovine fetuin glycoproteins. The ratios of relative intensity between the isotopically 2-AA labeled N-glycans were close to the theoretical molar ratios (1:1, 1:2, 1:5). We also demonstrated that the up-regulation of the Lewis antigen (∼82%) in sera from prostate cancer patients. In this proof-of-concept study, we demonstrated that the i-QTaG method, which enables to achieve a reliable comparative quantitation of total N-glycans via MALDI-TOF MS analysis, has the potential to diagnose and monitor alterations in glycosylation associated with disease states or biotherapeutics.

Original languageEnglish
Pages (from-to)840-848
Number of pages9
JournalBiotechnology Progress
Volume31
Issue number3
DOIs
StatePublished - 1 May 2015

Keywords

  • Comparative quantitation
  • MALDI-MS
  • N-glycan
  • Sialic acid neutralization
  • Stable isotopic labeling

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