Abstract
Celiac disease is an immune-mediated enteropathy precipitated by the ingestion of gluten-containing foods in genetically predisposed children and adults. After a positive diagnosis for celiac disease, the only available treatment is to adopt a gluten-free diet, and evaluation of the absence of gluten in foods is crucial for the health of celiac patients. In the present study, a recombinant glutamine-binding protein (GlnBP) from Escherichia coli showed its ability to recognize peptides deriving from digested wheat flour. GlnBP and the commercially available 4F3 monoclonal antibody, raised against a region of the α-gliadin peptide 33-mer from wheat, demonstrated the ability to detect gliadin extracted from wheat flour. Recombinant GlnBP and 4F3 monoclonal antibody were used as new capture agents for the development of a protein chip able to detect gluten in foods. The protein microarray system has proven to detect the presence of gliadin in a range of concentrations between 500 and 5 ppm.
Highlights
Celiac disease and wheat IgE-dependent allergy are diseases triggered by gluten present in wheat and by similar proteins in other grains such as rye, barley, kamut, spelt, and, in some cases, oats, and by non-gluten proteins [1]
By using total DNA extracted from E. coli cultured cells as template, the DNA amplified fragment, denoted as glutamine-binding protein (GlnBP), was cloned in the pET32 vector
The isolated GlnBP gene sequence consists of an open reading frame of 759 bp in length, which potentially codes for a protein of 249 amino acids with an estimated molecular mass of 27.4 kDa
Summary
Celiac disease and wheat IgE-dependent allergy are diseases triggered by gluten present in wheat and by similar proteins in other grains such as rye, barley, kamut, spelt, and, in some cases, oats, and by non-gluten proteins (e.g. amylase-trypsin inhibitors in bakers asthma) [1]. In order to protect celiac patients through a strict gluten-free diet, it is important to make early diagnosis, especially for the prevention of complications that could derive from celiac disease, such as autoimmune diseases (thyroiditis, diabetes, etc.), cancer (intestinal lymphoma) and other diseases with a strong social impact (fractures due to osteopenia, infertility, abortions, etc.) [4,5,6]. For these reasons, it is very important to promote help for celiac people through periodic checks of food sold as gluten-free, which may contain gluten contamination as a result of production processes. The Codex Alimentarius Commission allows a maximum gluten level of 20 ppm (mg/kg) in total for gluten-free food, based on the food as sold or distributed
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