Simultaneous determination of four food allergens using compact disc immunoassaying technology

Autores de IIS La Fe

• Sergi Morais Ezquerro

  Autor

• Ángel Maquieira Catalá

  Autor

Participantes ajenos a IIS La Fe

• Badran, Ahmed Ali

Grupos

• Unidad Mixta de Investigación en Nanomedicina y Sensores

Abstract

A multiplex competitive microimmunoassay for the simultaneous determination of gliadin, casein, beta-lactoglobulin, and ovalbumin is presented. The assay in microarray format is performed on a DVD where the allergens are physisorbed on the polycarbonate surface of the disc. The immunointeraction is detected using a mixture of specific gold-labeled antibodies and the signal amplified with the silver enhancement method. The optical density of the precipitate, read by a DVD drive, is related to the concentration of the four allergens in sample. An optimized protocol for the simultaneous extraction of the allergen proteins from food samples is also addressed. The suitability of the method is demonstrated for the simultaneous quantitative extraction and determination of the targeted allergens in spiked baby foods, juices, and beers. The sensitivity (EC50) of the multiplexed assay was 0.04, 0.40, 0.08, and 0.16 mg L-1 for gliadin, casein, beta-lactoglobulin, and ovalbumin, respectively, and the recovery results from the analysis of food samples ranged from 72 to 117%. A portable, easy-to-use, array-based bioanalytical method is developed for quantification of food allergens with a limit of detection below the accepted levels of the international legislations, which allows promotion of food safety and quality.

Keywords

• Food allergy; Gliadin; beta-lactoglobulin; Casein; Ovalbumin; Simultaneous extraction multiplex competitive microimmunoassay; Compact disc microarraying