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Immunological methods for allergen determination

ELISA is a common immunological method routinely used in analytical laboratories to detect and quantify allergens in food products. Antibodies are fundamental to ELISA as they are antigen (allergen)-specific and will determine the presence of an allergen in a sample extract.

As part of the objective to develop sensitive immunological methods, the main allergens of milk whey (ß-lactoglobulin), egg white (ovomucoid and ovalbumin) and egg yolk (a-livetin; from pasteurised and non-pasteurised yolks) were isolated and purified. Rabbits were immunised with purified proteins and sera was collected before immunisation (pre-bleed), during boost immunisations (test bleed) and after immunisation (terminal bleed).

Sera collected from the rabbits were characterised for antibody presence. The competitive direct ELISA technique, which allows binding of antibodies to protein antigens immobilised in wells of the microtitration plate was used for the investigations as well as competitive indirect ELISA.

Beside ELISA investigations the extracted proteins were separated electrophoretically, blotted and immuno stained. For the characterization of the patients' sera EAST was performed. EAST inhibition experiments were used for quantification of reduction of allergenic potency.

Reported by

Procter Department of Food Science, University of Leeds
Leeds LS2 9JT
LS2 9JT Leeds
United Kingdom
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