Tree nut and peanut allergies affect up to 1.4% US population. Upon exposure, sensitive individuals may experience adverse reactions, including fatal anaphylaxis, to tree nut and peanut allergens. Currently, there is no cure for food allergies. Strict avoidance of the offending allergens is the best defense for sensitive individuals. Consequently, it is important to develop reliable assays for food allergen detection. Several commercial polyclonal antibody (pAb)-based immunoassays are available for tree nut and peanut detection. However, the pAb-based assays often exhibit lack of specificity and robustness. The objective of the current investigation was to evaluate sensitivity, specificity, and robustness of monoclonal antibody (mAb)-based sandwich enzyme-linked immunosorbent assays (ELISAs) for almond, cashew, hazelnut, pecan, and peanut detection. The tested ELISAs are sensitive (limit of detections and limit of quantifications < 10 ppm) and reproducible (intra- and inter-assay variabilities < 24%). The laboratory developed ELISA used a mAb (4C10) targeting a well-defined amandin epitope that is recognized by several almond allergic patients’ sera IgE. Compared to the MonoTrace assay, the mAb 4C10-based ELISA was comparable in assay sensitivity and was superior in assay specificity and recognition of a human allergy relevant epitope. The detection mAbs targeted antigens were stable and detectable in whole tree nut and peanut seeds subjected to autoclaving, blanching, frying, microwaving, and dry roasting. No cross-reactivity was observed in 156 food ingredients, each tested at 100,000 ppm except for the hazelnut kit (cross-reactive with pecan and English walnut) and the pecan kit (cross-reactive with English walnut and black walnut). Antigen recovery ranges for spiked and incurred food matrices were 81‒126% and 22‒161%, respectively. The assay results were in agreement with the allergen declaration of 180 tested commercial foods with the exception of one sample where undeclared hazelnuts were detected, two samples where undeclared pecans were detected, and one sample where declared pistachio was not detected. In conclusion, the mAb-based ELISAs are sensitive, specific, and robust for tree nut and peanut detection and quantification under the tested conditions. / A Dissertation submitted to the Department of Nutrition, Food and Exercise Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Summer Semester 2016. / June 28, 2016. / ELISA, food allergy, monoclonal antibody, peanut, tree nut / Includes bibliographical references. / Shridhar K. Sathe, Professor Directing Dissertation; John G. Dorsey, University Representative; Yun-Hwa P. Hsieh, Committee Member; Kenneth H. Roux, Committee Member.
| Identifer | oai:union.ndltd.org:fsu.edu/oai:fsu.digital.flvc.org:fsu_366450 |
| Contributors | Liu, Changqi (authoraut), Sathe, Shridhar K. (professor directing dissertation), Dorsey, John G. (university representative), Hsieh, Peggy (committee member), Roux, Kenneth H. (committee member), Florida State University (degree granting institution), College of Human Sciences (degree granting college), Department of Nutrition, Food, and Exercise Science (degree granting department) |
| Publisher | Florida State University, Florida State University |
| Source Sets | Florida State University |
| Language | English, English |
| Detected Language | English |
| Type | Text, text |
| Format | 1 online resource (143 pages), computer, application/pdf |
| Rights | This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). The copyright in theses and dissertations completed at Florida State University is held by the students who author them. |
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