X6: A Novel Antibody for Potential Use in Gluten Quantification.

Yuri Lebedin,Ivan Shatalov,Aleksandrina Shatalova

doi:10.3390/molecules25143107

Yuri Lebedin, Ivan Shatalov + Show 1 more

Open Access

https://doi.org/10.3390/molecules25143107

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Abstract

Gliadin is a fraction of gluten, known to trigger celiac disease in susceptible people. To date, the life-long gluten-free diet is used for the prevention of this disease. Hence, methods for gluten control in foods are of significant importance. Being one of the most-used methods used for this purpose, ELISA should use high-affinity antibodies to gliadin peptides involved into celiac process. This study investigates the characteristics of a novel anti-gliadin antibody X6. We found the QXQPFPXP site to be a recognized epitope that provides specific binding of the antibody to cereal prolamins involved in celiac disease manifestation. A specificity study using immunoblotting shows the recognition of wheat, barley and rye proteins—as well as α-gliadin homologs from non-edible cereals (Dasypyrum villosum). Reactivity to avenin was less pronounced, as this protein does not contain the PFP motif most critical for antibody recognition. The proteins of Zea mays and Setaria italica were not recognized by X6. X6-based ELISA highly correlated with R5 and G12, which are Codex Alimentarius standards in the quantitative assessment of gluten content (Pearson’s R = 0.86 and 0.87, respectively). Qualitative assessment revealed no significant differences between R5 and G12 and X6.

Highlights

Celiac disease is a chronic inflammatory disorder of the small intestine and is mediated by gluten intake immune response in susceptible people [1]
Being one of the most-used methods used for this purpose, enzyme-linked immunosorbent assay (ELISA) should use high-affinity antibodies to gliadin peptides involved into celiac process
This study investigates the characteristics of a novel anti-gliadin antibody X6

Summary

Introduction

Celiac disease is a chronic inflammatory disorder of the small intestine and is mediated by gluten intake immune response in susceptible people [1]. Gliadins play a major role in the celiac disease manifestation. Scientists found the homologs of gliadin proteins in other cereals: in oats (avenin), barley (hordein), rye (secalin), and they are able to trigger the celiac disease process [5,6,7]. Most cases of celiac disease occur in patients with the HLA-DQ2/-DQ8 haplotype, which is the main risk-factor [8,9]. Researchers discovered the gliadin fragments deamidated by tissue transglutaminase (tTG) increase affinity for DQ2/-DQ8 heterodimeric surface receptors. The reaction of gliadin fragments with tTG is one of the key factors in the formation of the pathologic process [10]

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