1a. Objectives (from AD-416):
1. Develop simple model food systems to evaluate processing effects. 2. Assess the effects of various types of processing on allergenic foods, and the specific allergens contained therein, and methods for their detection using the model food systems.
1b. Approach (from AD-416):
1. Simple pastry squares (solid matrix) and liquid extracts of foods will be prepared with known amounts of commonly allergenic foods (milk, peanut). 2. We will apply various processing methods to the solid and liquid foods as appropriate, including boiling, baking, deep-fat frying, retorting, ultra-high temperature processing, microwaving, high pressure processing, radiofrequency processing, ultrasonic processing, pulsed electric field processing, acidification, proteolysis, and selected fermentations (specifically cheese as an example of fermentation of milk). 3. The effects of processing will be assessed using commercial immunoassay kits that detect either a mix of proteins from the allergenic source (e.g., total milk, total peanut) or a specific allergen (e.g., casein, ß-lactoglobulin, Ara h 1, Ara h 2, Ara h 3). Comparisons will also be made to human IgE-based immunoassays. 4. New approaches will be developed and comparatively evaluated to address assay deficiencies. New immunoassay approaches will be designed either (a) to detect pairs of epitopes on short peptides from specific allergens or (b) to use antisera prepared to process-modified antigens, or (c) to employ various alternative extraction techniques intended to resolubilize aggregated proteins, or (d) combinations of these approaches.
3. Progress Report:
The collaborative proposal by University of Nebraska and Agricultural Research Service has two main objectives. One is to develop simple model food systems to evaluate processing effects; and the second is to assess the effects of various types of processing on allergenic foods and the specific allergens contained therein, and methods for their detection using the model food systems. ARS has utilized allergen-specific antibodies and mass spectrometry to trace the individual allergens within various foods before and after processing to determine alterations. Overall, the project is progressing according to appropriate time line.