Application of enzyme-linked immunosorbent assay (ELISA) to wheat (Triticum aestivum L.) quality assessment

Abstract

Enzyme-linked immunosorbent assays (ELISAs), used for quantitative determination of the relative concentrations of antibody binding proteins, and two antibodies, specifically an anti-secalin monoclonal antibody (ASMab) and an anti-gliadin monoclonal antibody (AGMab), were used for wheat (Triticum aestivum L.) quality prediction. The anti-secalin monoclonal antibody (AGMab) was capable of identifying wheats carrying rye (Secale cereale L.) chromosome arm 1RS, in each of the 16 production environments, and also was able to discriminate among lines that carried 1AL/1RS, 1BL/1RS, or were heterogeneous for 1BL/1RS. When all genotypes (normal and 1RS wheats) were considered, negative correlations between the ELISA optical density variables and the quality parameters were observed. However, correlations between ELISA optical densities and quality parameters were actually positive among 1RS lines. This might be due to similar synthesis of secalin and wheat quality-related gluten proteins during grain filling period. The anti-gliadin monoclonal antibody (AGMab) was found to have high specificity for gamma-gliadin proteins. Immunoblot experiments, using Chinese Spring euploid and aneuploid lines, confirmed that AGMab bound primarily to gliadins encoded by the short arm of chromosome 1A. Few significant correlations were observed between AGMab ELISA optical density parameters and quality parameters, among all genotypes. However, within specific genotypes, significant correlations were detected with several quality parameters. This suggests that use of AGMab for the prediction of quality would be limited to certain responsive genotypes

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