Determination of B-cell epitopes in patients with celiac disease: Peptide microarrays

Rok Seon Choung, Eric V. Marietta, Carol T. Van Dyke, Tricia L. Brantner, John Rajasekaran, Pankaj J. Pasricha, Tianhao Wang, Kang Bei, Karthik Krishna, Hari K. Krishnamurthy, Melissa R. Snyder, Vasanth Jayaraman, Joseph A. Murray

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Background Most antibodies recognize conformational or discontinuous epitopes that have a specific 3- dimensional shape; however, determination of discontinuous B-cell epitopes is a major challenge in bioscience. Moreover, the current methods for identifying peptide epitopes often involve laborious, high-cost peptide screening programs. Here, we present a novel microarray method for identifying discontinuous B-cell epitopes in celiac disease (CD) by using a silicon-based peptide array and computational methods. Methods Using a novel silicon-based microarray platform with a multi-pillar chip, overlapping 12-mer peptide sequences of all native and deamidated gliadins, which are known to trigger CD, were synthesized in situ and used to identify peptide epitopes. Results Using a computational algorithm that considered disease specificity of peptide sequences, 2 distinct epitope sets were identified. Further, by combining the most discriminative 3-mer gliadin sequences with randomly interpolated3- or 6-mer peptide sequences, novel discontinuous epitopes were identified and further optimized to maximize disease discrimination. The final discontinuous epitope sets were tested in a confirmatory cohort of CD patients and controls, yielding 99% sensitivity and 100% specificity. Conclusions These novel sets of epitopes derived from gliadin have a high degree of accuracy in differentiating CD from controls, compared with standard serologic tests. The method of ultrahigh- density peptide microarray described here would be broadly useful to develop highfidelity diagnostic tests and explore pathogenesis.

Original languageEnglish (US)
Article numbere0147777
JournalPloS one
Issue number1
StatePublished - Jan 1 2016

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General


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