Abstract
Objectives
To develop a sensitive and quantitative method for monitoring the abnormal glycosylation of clinical and biopharmaceutical products.
Results
MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) was proposed for sensitive and quantitative analysis of total N-glycans. The derivatization reactions (i.e., amidation of sialic acid and incorporation of a positive charge moiety into the reducing end) dramatically increased the linearity (R2 > 0.99) and sensitivity (limit of detection is 0.5 pmol/glycoprotein) relative to underivatized glycans. In addition, the analytical strategy was chromatographic purification-free and non-laborious process accessible to the high-throughput analyses. We used MALDI-QTaG to analyze the N-glycans of α-fetoprotein (AFP) purified from normal cord blood and HCC cell line (Huh7 cells). The total percentages of core-fucosylated AFP N-glycans from Huh7 cells and normal cord blood were 98 and 18 %, respectively.
Conclusions
This MALDI-MS-based glycomics technology has wide applications in many clinical and bioengineering fields requiring sensitive, quantitative and fast N-glycosylation validation.
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Acknowledgments
This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) (NRF-2013R1A1A1004998) and Polar Academic Program (PAP, PD13010) from KOPRI.
Supporting information
Supplementary Table 1—Comparison between relative intensities of the N-glycans derived from AFP of human cord serum and Auh7 cell labeled with Ah and GP obtained by MALDI-MS
Supplementary Figure 1—Determination of limit of detection (LOD) for N-glycans from bovine fetuin using MALDI-QTaG method (0.5 pmol on the MALDI spot)
Supplementary Figure 2—Comparison of relative quantities of N-glycans derived from human IgG using MALDI-MS and UPLC
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Kim, KJ., Kim, YW., Hwang, CH. et al. A MALDI-MS-based quantitative targeted glycomics (MALDI-QTaG) for total N-glycan analysis. Biotechnol Lett 37, 2019–2025 (2015). https://doi.org/10.1007/s10529-015-1881-6
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DOI: https://doi.org/10.1007/s10529-015-1881-6