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Soybean allergies are one of top eight types of food allergies, affecting ~ 0.4% children and 0.3% adults in North America. Soybean agglutinin (SBA) protein, also known as soybean lectin, is both an allergen as well as an anti-nutrient that reducing the nutritional value of soybean and soybean products. The objective of this study was to obtain a validated methodology that is precise and accurate in the measurement of SBA while allowing minimally equipped laboratories to effectively conduct the analysis. A competitive ELISA was constructed and optimized by using rabbit anti whole soybean polyclonal antibodies (pAb) as primary Ab. The constructed competitive ELISA was validated to have LOD of 0.063 ng/ml of soybean lectin in all tested samples. The ELISA is reproducible and accurate as CVs of all ELISAs tested were less than 24% and the average recoveries were within 15% of the actual value, which demonstrating accuracy of the assay. However, further investigate is needed to evaluating CV of the assay. The validated ELISA method was able to detect and quantify the SBA in 20 soybean varieties and indicated that the natural variability of SBA is subject to the effects of genotype and environment. Moreover, the ELISA can detect the SBA content in thermal processed soybean, germinated soybean sprouts, and commercial soybean products. The results suggested that processing methods can affect soybean lectin immunoreactivity. Compare to the conventional hemagglutinating assay, ELISA is more sensitive and effective. In conclusion, the constructed pAb-based competitive ELISA provides an efficacious detection and quantification for the soybean lectin.
A Thesis submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Master of Science.
Includes bibliographical references.
Shridhar K. Sathe, Professor Directing Thesis; Qinchun Rao, Committee Member; John G. Dorsey, Committee Member.
Florida State University
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