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

PloS One
Rok Seon ChoungJoseph A Murray

Abstract

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%...Continue Reading

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Citations

Feb 9, 2019·Biomarkers : Biochemical Indicators of Exposure, Response, and Susceptibility to Chemicals·Thushani SiriwardhaneHari Krishnamurthy
Jul 7, 2018·Biomarker Insights·Yuanyuan YangHari Krishnamurthy
Dec 19, 2016·Clinical & Translational Immunology·Melinda Y Hardy, Jason A Tye-Din
May 7, 2021·Journal of the American Society of Nephrology : JASN·Laith Farah Al-RabadiLaurence H Beck

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Methods Mentioned

BETA
Atomic force microscopy
chips
deamidation
the
chip
peptide array
biopsy
enzyme-linked immunosorbent assay
ELISA
blood

Software Mentioned

MATLAB

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