Faculty of Food Biotechnologies and Engineering, ITMO University, Lomonosova Street, 9, 191002 St. Petersburg, Russia.
ELISA Development Department, XEMA Co., Ltd., Degtyarniy Lane, 8-10 St., 191144 Petersburg, Russian.
Molecules. 2020 Jul 8;25(14):3107. doi: 10.3390/molecules25143107.
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 (). Reactivity to avenin was less pronounced, as this protein does not contain the PFP motif most critical for antibody recognition. The proteins of and 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.
麦醇溶蛋白是面筋的一个组成部分，已知它会引发乳糜泻。迄今为止，人们一直采用终生无麸质饮食来预防这种疾病。因此，控制食物中麸质的方法非常重要。酶联免疫吸附试验（ELISA）是目前最常用的方法之一，应使用高亲和力的针对乳糜泻相关的麦醇溶蛋白肽的抗体。本研究调查了新型抗麦醇溶蛋白抗体 X6 的特性。我们发现 QXQPFPXP 位点是一个公认的抗原表位，该抗体能够特异性地与参与乳糜泻发病的谷物醇溶蛋白结合。免疫印迹特异性研究表明，该抗体能够识别小麦、大麦和黑麦蛋白，以及非食用谷物中的α-麦醇溶蛋白同源物。然而，对燕麦醇溶蛋白的反应性则不那么明显，因为这种蛋白不含有对于抗体识别至关重要的 PFP 基序。X6 不能识别 和 的蛋白。X6 基于 ELISA 的方法与 Codex Alimentarius 定量评估麸质含量的标准 R5 和 G12 高度相关（Pearson 相关系数分别为 0.86 和 0.87）。定性评估显示，R5 和 G12 与 X6 之间没有显著差异。
