Analytical and Bioanalytical Chemistry

, Volume 410, Issue 6, pp 1617–1629 | Cite as

Designation of fingerprint glycopeptides for targeted glycoproteomic analysis of serum haptoglobin: insights into gastric cancer biomarker discovery

  • Jua Lee
  • Serenus Hua
  • Sung Hyeon Lee
  • Myung Jin Oh
  • Jaekyung Yun
  • Jin Young Kim
  • Jae-Han Kim
  • Jung Hoe Kim
  • Hyun Joo An
Paper in Forefront


Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide, largely because of difficulties in early diagnosis. Despite accumulating evidence indicating that aberrant glycosylation is associated with GC, site-specific localization of the glycosylation to increase specificity and sensitivity for clinical use is still an analytical challenge. Here, we created an analytical platform with a targeted glycoproteomic approach for GC biomarker discovery. Unlike the conventional glycomic approach with untargeted mass spectrometric profiling of released glycan, our platform is characterized by three key features: it is a target-protein-specific, glycosylation-site-specific, and structure-specific platform with a one-shot enzyme reaction. Serum haptoglobin enriched by immunoaffinity chromatography was subjected to multispecific proteolysis to generate site-specific glycopeptides and to investigate the macroheterogeneity and microheterogeneity. Glycopeptides were identified and quantified by nano liquid chromatography–mass spectrometry and nano liquid chromatography–tandem mass spectrometry. Ninety-six glycopeptides, each corresponding to a unique glycan/glycosite pairing, were tracked across all cancer and control samples. Differences in abundance between the two groups were marked by particularly high magnitudes. Three glycopeptides exhibited exceptionally high control-to-cancer fold changes along with receiver operating characteristic curve areas of 1.0, indicating perfect discrimination between the two groups. From the results taken together, our platform, which provides biological information as well as high sensitivity and reproducibility, may be useful for GC biomarker discovery.

Graphical abstract


Mass spectrometry Glycoproteomics Site-specific glycosylation Glycopeptide Gastric cancer 



This work was supported by Korea Basic Science Institute grant (T37413) and by the Institute for Basic Science (IBS-R001-D1-2017-a00).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participat

Human sera were collected by the National Biobank of Korea by means of a standardized protocol approved by the Ethics Committees of Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea (KH2010-15). Informed consent was provided by all individuals involved in the study.

Supplementary material

216_2017_811_MOESM1_ESM.pdf (851 kb)
ESM 1 (PDF 850 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Jua Lee
    • 1
    • 2
  • Serenus Hua
    • 1
    • 2
  • Sung Hyeon Lee
    • 3
  • Myung Jin Oh
    • 1
    • 2
  • Jaekyung Yun
    • 1
    • 2
  • Jin Young Kim
    • 4
  • Jae-Han Kim
    • 5
  • Jung Hoe Kim
    • 6
  • Hyun Joo An
    • 1
    • 2
  1. 1.Asia Glycomics Reference SiteChungnam National UniversityDaejeonRepublic of Korea
  2. 2.Graduate School of Analytical Science and Technology, #455 College of Engineering IIChungnam National UniversityDaejeonRepublic of Korea
  3. 3.GLYCAN Co. Ltd.Healthcare Innovation ParkSeongnamRepublic of Korea
  4. 4.Department of Mass SpectrometryKorea Basic Science InstituteOchangRepublic of Korea
  5. 5.Department of Food and NutritionChungnam National UniversityDaejeonRepublic of Korea
  6. 6.Department of Biological SciencesKorea Advanced Institute of Science and TechnologyDaejeonRepublic of Korea

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