Standardization of Food Allergen Measurements Using Multiplex Array Technology.

Multiplex suspension array technology enables multiple protein analytes to be measured in a single assay. In this chapter, we describe a polystyrene bead-based fluorescent multiplex array for quantitative measurements of specific food allergens. The array uses monoclonal antibodies for allergen detection and purified allergens as reference standards.

Simultaneous quantification of specific food allergen proteins using a fluorescent multiplex array.

The aim was to develop a fluorescent multiplex array for simultaneously measuring regulated food allergens using specific allergen protein molecules from peanut, tree nut, cow's milk, egg, soy, fish, shellfish, sesame, mustard and celery. Microspheres coupled to specific monoclonal antibodies were used for allergen detection, with purified allergens as reference standards.Standard curves for 17 allergens covered a 5-log dynamic range.

Detection of Food Allergens in School and Home Environments of Elementary Students.

Background: Little is known about environmental food allergen exposure on school surfaces.

Objective: To compare the distribution of major food allergens in floor dust and table wipe samples from elementary schools and dust samples from students' homes.

Methods: In this substudy of the School Inner-City Asthma Study-II, 103 table wipe samples and 98 floor dust samples from cafeterias and classrooms in 18 elementary schools were analyzed for milk, peanut, cashew, hazelnut, and egg using a multiplex array.

Doses of Specific Allergens in Early Introduction Foods for Prevention of Food Allergy.

Background: Consumption of common allergenic foods, such as peanut, in early life can reduce the risk of food allergy among high-risk children and is recommended in revised clinical guidelines. Commercial early allergen introduction foods (EIF) containing single or multiple allergenic foods for feeding infants are promoted to consumers and health care providers as aids to prevent food allergy.

Objective: To determine the concentration and doses of major food allergens in EIF.

Variability in German Cockroach Extract Composition Greatly Impacts T Cell Potency in Cockroach-Allergic Donors.

German cockroach extract is used clinically to evaluate allergen-specific sensitization and for subcutaneous allergen-specific immunotherapy, though there are no guidelines for standardization in its manufacture. We performed an immunological evaluation of 12 different cockroach extracts prepared from different sources and their potency to induce allergen-specific T cell reactivity. PBMC from 13 cockroach allergic donors were expanded with 12 different German cockroach extracts.

Allergen content in German cockroach extracts and sensitization profiles to a new expanded set of cockroach allergens determine in vitro extract potency for IgE reactivity.

Background: Cockroach allergens are an important cause of IgE-mediated sensitization in inner-city asthmatic patients. However, cockroach extracts used for diagnosis and immunotherapy are not standardized.

Objective: We sought to determine the allergen content of nonstandardized German cockroach extracts and the levels of sensitization to an expanded set of cockroach allergens as determinants of in vitro extract potency for IgE reactivity.

Specific allergen profiles of peanut foods and diagnostic or therapeutic allergenic products.

Background: Generic immunoassays for peanut cannot discriminate between allergen levels in peanut-derived food products or therapeutics. Clinical trials of oral immunotherapy (OIT) are strengthened by using standardized peanut preparations with defined doses of major allergens.

Objective: This article describes measurement of Ara h 1, Ara h 2, and Ara h 6 in peanut foods and in peanut flour extracts used for allergy diagnosis and OIT.

A multi-center ring trial of allergen analysis using fluorescent multiplex array technology.

Background: Consistent performance of allergen assays is essential to ensure reproducibility of exposure assessments for investigations of asthma and occupational allergic disease. This study evaluated intra- and inter-laboratory reproducibility of a fluorescent multiplex array, which simultaneously measures eight indoor allergens in a single reaction well.

Methods: A multi-center study was performed in nine laboratories in the US and Europe to determine the inter-laboratory variability of an 8-plex array for dust mite, cat, dog, rat, mouse and cockroach allergens.