Details

Autor(en) / Beteiligte

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

Titel

Determination of B-Cell Epitopes in Patients with Celiac Disease: Peptide Microarrays

Ist Teil von

• PloS one, 2016-01, Vol.11 (1), p.e0147777-e0147777

Ort / Verlag

United States: Public Library of Science

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. 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. 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. 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 ultra-high-density peptide microarray described here would be broadly useful to develop high-fidelity diagnostic tests and explore pathogenesis.