APPLICATION OF WESTERN BLOT ANALYSIS FOR DETECTION OF PROLAMIN PROTEINS IN CEREAL GRAINS AND BREAD

摘要

Celiac disease is an inflammatory condition of the small intestine in genetically susceptible individuals caused by ingestion of wheat gluten and corresponding proteins from barley and rye. Cereal storage proteins (prolamins) are responsible for immunological response of patients with celiac disease. Prolamins are alcohol soluble fractions, namely gliadins (wheat), hordeins (barley) and secalins (rye). The main triggering factor is wheat fraction with low molecular weight (20-30 kDa) called α-gliadins. Immunochemical detection of celiac active proteins is based on reactivity of gluten-detecting antibodies with prolamins extracted from cereals. In our study, we used Western blot analysis for detection of prolamin complex in cereal grains and processed foods (breads). Western blot was carried out by polyclonal antibody raised against wheat gluten. Reaction was positive for all kind of cereal grains. The samples of wheat and spelt wheat show much more positive affinity to antibody than rye and oat. As well as for cereal grains, all samples of bread showed positive immunological reaction with used antibody. Western blot analysis with gluten polyclonal antibody is suitable method for qualitative detection of prolamin complex in cereal grains and processed foods.