Celiac Disease-specific Prolamin Peptide Content of Wheat Relatives and Wild Species Determined by ELISA Assays and Bioinformatics Analyses

摘要

Enzyme-linked immunosorbent assays (ELISAs) are widely used to determine gluten contamination\udin gluten-free and low gluten food samples. ELISA assays developed using\udmonoclonal antibodies against known toxic peptides have an advantage in the identification of\udtoxic prolamin content in protein extracts of different food samples, as well as raw materials.\udR5 and G12 monoclonal antibodies specific for two known toxic peptides used in commercially\udavailable gluten ELISA assays were applied to test toxic peptide contents in wheat relatives\udand wild wheat species with different genome composition and complexity. Although the\udR5 peptide content showed some correlation with ploidy levels in Triticum species, there was a\udhigh variance among Aegilops species. Some of the analysed diploid Aegilops species showed\udextremely high R5 peptide contents. Based on the bioinformatics analyses, the R5 peptide was\udpresent in most of the sulphur rich prolamins in all the analysed species, whereas the G12\udepitope was exclusively present in alpha gliadins. High variation was detected in the position\udand frequency of epitopes in sequences originating from the same species, thus highlighting\udthe importance of genotypic variation within species. Identification of new prolamin alleles of\udwheat relatives and wild wheat species is of great importance in order to find germplasm for\udspecial end-use quality purposes as well as development of food with reduced toxicity.