durum do not contain the D genome, are hence already devoid of the known immunogenic sequences of -gliadins [11]. of relatively small population of genetically predisposed individuals, who under this toxic action develop celiac disease (CD). As the quantity of immunogenic gliadin could vary between different wheat species, the first aim of this work was to determine the percentage of immunogenic gliadin in ten bread wheat Dihydrokaempferol cultivars and in three commercially grown durum wheat cultivars. The second part of the study was initiated by results of previous publication, reporting that sera of some of Dihydrokaempferol multiple myeloma (MM) patients showed the presence of elevated levels of anti-gliadin IgA, without the enhanced levels of anti-gliadin IgG antibodies, decided with commercial ELISA test. It was designed to assess is it possible to reveal is there any hidden, especially anti-gliadin IgG immunoreactivity, in serum of mentioned group of patients. For this purpose we tested MM patients sera, as well as celiac disease (CD) patients sera for the immunoreaction with the native gliadin isolated from wheat Dihydrokaempferol species used for bread and pasta making in corresponding geographic region. Results Gliadin was isolated from wheat flour by two step 60% ehanolic extraction. Its content was determined by commercial R5 Mendez Elisa using PWG gliadin as the standard. Results obtained showed that immunogenic gliadin content varies between 50.4 and 65.4 mg/g in bread wheat cultivars and between 20 and 25.6 mg/g in durum wheat cultivars. Anti-gliadin IgA and IgG immunoreactivity of patients’ sera in (IU/ml) was firstly determined by commercial diagnostic Binding Site ELISA test, and then additionally by non-commercial ELISA assessments, using standardized ethanol wheat extracts -gliadin as the antigen. In both patients groups IgA immunoreactivity to gliadin from different cultivars was almost homogenous PYST1 and in correlation with results from commercial test (except for one patient with IgA() myeloma, they were more then five times higher). But, results for IgG immunoreactivity were more frequently inhomogeneous, and especially for few MM patients, they were more then five times higher and did not correlate with results obtained using Binding Site test. Conclusion Results obtained showed different content of immunogenic gliadin epitopes in various species of wheat. They also point for new effort to elucidate is there a need to develop new standard antigen, the representative mixture of gliadin isolated from local wheat species used for bread production in corresponding geographic region for ELISA diagnostic assessments. Background It is well known that gliadin is usually directly or indirectly through immune mediated reactions, toxic to small bowel mucosa of relatively small population of genetically predisposed individuals who under this toxic action develop celiac disease (CD). These patients need to eat food without gluten, i.e., they need to be on gluten free diet (GFD). Therefore very reliable assessments are needed to determine is the content of gliadin really below the accepted value (20 mg/kg). As gliadin isolated from various species used as the antigen may have different immunogenicity [1] that fact could be a problem in the immunological assessments used for determination of gliadin content in food; i.e., results may greatly depend on the origin and type of gliadin that was used for calibration. In the Dihydrokaempferol aim to overcome this analytical problem, “prolamin working group” developed a PWG gliadin which represents protein fraction soluble in 60% ethanol from the mixture of twenty-eight wheat cultivars grown in Great Britain, France and Germany [1-6]. This gliadin is recommended for use as the standard antigen in immunological techniques for determination of gluten content in food. At the same time, it was very important to standardize anti-gliadin antibodies that should be used in immunological assessments for determination of gliadin content. In the recent time few monoclonal or polyclonal anti-gliadin antibodies were developed. Commercial kits often used polyclonal antibodies developed against wheat gliadin, or a monoclonal antibody made against -gliadins from the Australian wheat cultivar ‘Timgalen’. There were also other antibodies developed in different laboratories such as monoclonal antibody PN3, raised against a synthetic peptide equivalent to the amino acids 31C49 of -gliadin, i.e. the sequence of toxic peptide of -gliadin, which has been shown to.