Research Article

Comparative proteomic analysis of de-N-glycosylated serum from hepatitis B carriers reveals polypeptides that correlate with disease status

  1. Mary Ann Comunale1,
  2. Taj S. Mattu1,
  3. Melissa A. Lowman1,
  4. Alison A. Evans2,
  5. W. Thomas London2,
  6. O. John Semmes3,
  7. Michael Ward3,
  8. Richard Drake3,
  9. Patrick R. Romano1,
  10. Laura F. Steel1,
  11. Timothy M. Block1,
  12. Anand Mehta1,*

Article first published online: 15 DEC 2003

DOI: 10.1002/pmic.200300625

Issue

PROTEOMICS

PROTEOMICS

Volume 4, Issue 3, pages 826–838, March 2004

Additional Information

How to Cite

Comunale, M. A., Mattu, T. S., Lowman, M. A., Evans, A. A., London, W. T., Semmes, O. J., Ward, M., Drake, R., Romano, P. R., Steel, L. F., Block, T. M. and Mehta, A. (2004), Comparative proteomic analysis of de-N-glycosylated serum from hepatitis B carriers reveals polypeptides that correlate with disease status. PROTEOMICS, 4: 826–838. doi: 10.1002/pmic.200300625

Author Information

  1. 1

    Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, The Jefferson Center, Doylestown, PA, USA

  2. 2

    Fox Chase Cancer Center, Philadelphia, PA, USA

  3. 3

    Microbiology and Immunology Department, Eastern Virginia Medical School, Norfolk, VA, USA

*The Jefferson Center, Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, 700 East Butler Avenue, Doylestown, PA, USA, 18901 Fax: +1-215-489-4920

Publication History

  1. Issue published online: 19 FEB 2004
  2. Article first published online: 15 DEC 2003
  3. Manuscript Received: 14 APR 2003

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Keywords:

  • Diagnostic markers;
  • Glycosylation;
  • Hepatitis B;
  • Hepatocellular carcinoma;
  • Two-dimensional gel electrophoresis

Abstract

Analysis of the polypeptide profile in tissues, cells, and sera by high-resolution two-dimensional (2-D) gel electrophoresis offers promise in the identification of biomarkers that correlate with disease. However, sera contain many polypeptides bearing N-linked glycosylation that can complicate interpretation. Therefore, we tested the possibility that de-N-glycosylation of the polypeptides present in human serum would result in a simplification of serum proteome profiles. Briefly, polypeptides present in human serum were left untreated or subjected to de-N-glycosylation by incubation with PNGase F and resolved by high-resolution 2-D gel electrophoresis. De-N-glycosylation reduced the number of glycoform variants, enhanced the resolution of many polypeptides and allowed other polypeptides to become visible. As an initial test of concept, clinically relevant serum samples from individuals with or without diagnosis of hepatocellular carcinoma were compared. Several polypeptides, apparent only after de-N-glycosylation, were shown to correlate with disease. Although the results are preliminary and the identities of all the putative biomarkers not yet known, the data suggest that de-N-glycosylation offers a method to enhance the resolution of serum polypeptide profiles and has value in comparative proteomic studies.

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