Stable Isotopic Labeling-Based Quantitative Targeted Glycomics (i-QTaG)
- Authors
- Kim, Kyoung-Jin; Kim, Yoon-Woo; Kim, Yun-Gon; Park, Hae-Min; Jin, Jang Mi; Kim, Young Hwan; Yang, Yung-Hun; Lee, Jun Kyu; Chung, Junho; Lee, Sun-Gu; Saghatelian, Alan
- Issue Date
- May-2015
- Publisher
- WILEY-BLACKWELL
- Keywords
- MALDI-MS; N-glycan; comparative quantitation; stable isotopic labeling; sialic acid neutralization
- Citation
- BIOTECHNOLOGY PROGRESS, v.31, no.3, pp.840 - 848
- Journal Title
- BIOTECHNOLOGY PROGRESS
- Volume
- 31
- Number
- 3
- Start Page
- 840
- End Page
- 848
- URI
- http://scholarworks.bwise.kr/ssu/handle/2018.sw.ssu/8746
- DOI
- 10.1002/btpr.2078
- ISSN
- 8756-7938
- 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 (13) C-6/ (12) C-6-2-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 (R-2 > 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 (similar to 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. (C) 2015 American Institute of Chemical Engineers
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