Verification of the Gluten Presence Using the Electrophoresis Technique SDS-PAGE in Gluten-Free Breads Prepared to Celiac Patients in Isfahan City

Celiac disease or gluten-sensitive enteropathy is an immune-related disease which is a permanent sensitivity to wheat Geliadin or barely Prolamine in people who are genetically predisposed. Gluten is a protein found in wheat, barley and rye. This substance causes the stretching of the dough and crunchy texture of the final product. Gluten is composed of two subunits, Gliadin and Glutenin. The Geliadin is alcohol-soluble and Glutenin is just soluble in dilute acids and bases and based on these features Geliadin can be isolated. The detection of gluten in gluten-free bread flour can verify the manufacturers` claims on these products and also comparison the gluten free breads with ordinary ones.
In this study, Gel electrophoresis technique" SDS-PAGE" was used to detect and measure the protein gluten. A total of 24 samples of flour and alcohol Iso-propanol was used as the alcoholic solvent. The solubility in alcohol, protein Geliadin in the presence of the sample will be extracted. In the next step by electrophoresis, extracted composition after preparation buffer and a marker protein, as well as pure Gluten as a positive control, the Acrylamide gel was injected. Upon completion of this step, the gel solution Coomassie Blue colored paint and then paints the solution was clear.
Findings: In the study gel obtained from this test compared with the positive control sample as well as the standard marker proteins, Geliadin bands within the gel was invisible, indicating the absence of gluten in the samples of flour.
The electrophoresis technique SDS-PAGE for high sensitivity, the poor amounts of gluten can be measured, so the incidence of clinical protests in Celiac disease will be prevented. Another benefit of this technique is its high speed and low cost. Other techniques have been developed to measure and extract the gluten in previous years, including ELISA, Dot- immunobinding assay method using antibodies to Gluten, as well as a relatively high sensitivity PCR these methods also are entitled. Since electrophoresis SDS-PAGE method has not been used in Iran to qualitative determination of Gluten in bread, for the first time as preventive screening tests for the isolation of gluten-free flour produced in the country are presented.