Optimization of a competitive ELISA with polyclonal antibodies for quantification of prolamins in foods

Abstract
The optimization of a sequential competitive ELISA for the quantification of prolamins in foods is described in this article. The assay was developed using polyclonal antibodies obtained by the hyper‐immunization of rabbits with commercial gliadin. The ELISA developed in this way showed a very high degree of detectability (detection limit, 1 ng ml‐1), as well as the ability to discriminate between prolamins harmful to coeliac individuals from non‐toxic prolamins. The influence of the solvent used for extraction of the samples on the detection capability of the test was also studied. The assay proved to be useful for the evaluation of gliadins in processed foods including meat products. The assay was applied to many types of foods and was compared with a commercial kit approved by the Association of Official Analytical Chemists.

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