Mass spectrometry-based seed allergen quantification and applications in seed quality assessment
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Incidences of food allergies have doubled in the past two decades, becoming a worldwide public health concern. Together, tree nuts and peanuts cause approximately 80% of anaphylactic reactions and account for over 50% of child fatalities related to food allergies. For this reason, accurate and sensitive methods are necessary to identify and quantify seed allergens as a means to provide consumer information on seed and seed-derived food-products. New technologies (e.g. proteomics and mass spectrometry) can provide the means to compare allergen concentrations among seed varieties to evaluate the contribution of both genetic and environmental factors. The work presented, describes the development of quantitative mass spectrometry techniques and their application to the study of seed composition dynamics. Firstly, the effects of RNAi-mediated reduction of major seed allergens (Ara h 2 and 6) on seed protein composition were studied using spectral counting and the Absolute QUAntitation strategy (AQUA). Using these methods, the available proteome was surveyed for major alterations in both on-target (Ara h 2 and 6) and potential off-target proteins. Several off-target alterations were discovered including an increase in another known allergen. Secondly, the AQUA strategy was further applied to study the effects of environment and genotype on allergens and anti-nutritional proteins in soybean. In this study we analyzed the levels of known allergens and anti-nutritional proteins in commercially available soybean lines grown in six locations spanning three climate zones in North America. Robust statistical analysis shows that the effects of environment far outweigh those of genotype.--From public.pdf
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