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Clinical Chemistry 53: 1254-1263, 2007. First published May 17, 2007; 10.1373/clinchem.2007.085563
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Right arrow Proteomics and Protein Markers
(Clinical Chemistry. 2007;53:1254-1263.)
© 2007 American Association for Clinical Chemistry, Inc.

Proteomics and Protein Markers

High-Throughput Quantitative Profiling of Serum N-Glycome by MALDI-TOF Mass Spectrometry and N-Glycomic Fingerprint of Liver Fibrosis

Richard K.T. Kam1,2, Terence C.W. Poon1,2,3,a, Henry L.Y. Chan2,3, Nathalie Wong4, Alex Y. Hui2 and Joseph J.Y. Sung2,3

1 Li Ka Shing Institute of Health Sciences, 2 Department of Medicine and Therapeutics, 3 Centre for Emerging Infectious Diseases, and 4 Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong SAR.

aAddress correspondence to this author at: Li Ka Shing Institute of Health Sciences, Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, N.T., Hong Kong SAR. Fax 852-2648-8842; e-mail tcwpoon{at}cuhk.edu.hk.

Background: The use of MALDI-TOF mass spectrometry (MS) in quantitative glycan profiling has not been reported. In this study, we attempted to establish a high-throughput quantitative assay for profiling serum N-glycome, and we applied the new assay to identifying serum N-glycans for diagnosis of liver fibrosis and cirrhosis.

Methods: N-glycans from whole serum proteins in 2 µL serum were released by enzymatic digestion, cleaned up by hydrophilic chromatography, and subsequently quantitatively profiled with a linear MALDI-TOF MS system, which was originally designed for quantitative proteomic profiling. Serum N-glycome profiles from 46 patients with chronic hepatitis B infection and with different degrees of liver fibrosis were examined.

Results: The intra- and interassay CVs of peak intensities of the standard N-glycans were <8% and <17%, respectively. When the assay was applied to the analysis of serum N-glycome profiles, 17 peaks were found to be potential biomarkers for detection of liver fibrosis/cirrhosis. Linear regression analysis revealed that 4 peaks of 1341.5, 1829.7, 1933.3, and 2130.3 m/z (all P <0.005) had complementary value in detecting liver fibrosis and included them, but not any serological markers, in the diagnostic model. Leave-one-out cross-validation showed the diagnostic model could identify significant fibrosis (Ishak score ≥3) and cirrhosis (Ishak score ≥5), both at 85% accuracy.

Conclusion: This is the first study to illustrate the quantitative aspect of MALDI-TOF MS in N-glycome profiling and the first study to reveal the potential value of the serum N-glycan profile for identifying liver fibrosis.




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