A high-throughput O-glycopeptide discovery platform for seromic profiling

J Proteome Res. 2010 Oct 1;9(10):5250-61. doi: 10.1021/pr1005229.


Biomarker microarrays are becoming valuable tools for serological screening of disease-associated autoantibodies. Post-translational modifications (PTMs) such as glycosylation extend the range of protein function, and a variety of glycosylated proteins are known to be altered in disease progression. Here, we have developed a synthetic screening microarray platform for facile display of O-glycosylated peptides (O-PTMs). By introduction of a capping step during chemical solid-phase glycopeptide synthesis, selective enrichment of N-terminal glycopeptide end products was achieved on an amine-reactive hydrogel-coated microarray glass surface, allowing high-throughput display of large numbers of glycopeptides. Utilizing a repertoire of recombinant glycosyltransferases enabled further diversification of the array libraries in situ and display of a new level of potential biomarker candidates for serological screening. As proof-of-concept, we have demonstrated that MUC1 glycopeptides could be assembled and used to detect autoantibodies in vaccine-induced disease-free breast cancer patients and in patients with confirmed disease at time of diagnosis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantibodies / blood*
  • Autoantibodies / immunology
  • Biomarkers, Tumor / blood*
  • Biomarkers, Tumor / immunology
  • Breast Neoplasms / blood
  • Breast Neoplasms / diagnosis
  • Breast Neoplasms / immunology
  • Cancer Vaccines / immunology
  • Female
  • Glycopeptides / analysis*
  • Glycopeptides / immunology
  • Glycopeptides / metabolism
  • Glycosylation
  • Glycosyltransferases / metabolism
  • Humans
  • Mucin-1 / analysis
  • Mucin-1 / immunology
  • Mucin-1 / metabolism
  • Peptides / analysis
  • Peptides / immunology
  • Peptides / metabolism
  • Proteomics / methods*


  • Autoantibodies
  • Biomarkers, Tumor
  • Cancer Vaccines
  • Glycopeptides
  • MUC1 protein, human
  • Mucin-1
  • Peptides
  • Glycosyltransferases