Magnetic particles functionalized with PAMAM-dendrimers and antibodies: a new system for an ELISA method able to detect Ara h3/4 peanut allergen in foods |
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Authors: | Francesca Speroni Lisa Elviri Maria Careri Alessandro Mangia |
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Institution: | 1. Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Viale G.P. Usberti 17/A, 43100, Parma, Italy
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Abstract: | An innovative enzyme-linked immunosorbent assay (ELISA) format based on antibody-coated magnetic micro-particles (MPs) for
the sensitive detection of Ara h3/4 allergen in food is described. The immunosupport is suspended in the incubation solutions
and the MPs with the captured allergen can be easily harvested on a magnet, separated from the solutions, and washed using
an easy-to-use, fast and selective approach that allows its detection and quantification. Two differently coated MPs, ProteinA-Pn-b
and MP-NH2-PAMAM G 1.5 -Pn-b immunosupports, were tested. The functionalization of the MPs with PAMAM-sodium carboxylate dendrimers
elicits a major stability on the immunoglobulin activity resulting in a threefold enhancement of the analytical sensitivity
for the assay with respect to a ProteinA immobilization. Validation was carried out on two different matrices: corn flakes
and biscuits. In the case of MP-NH2-PAMAM G 1.5 -Pn-b immunosupport, limit of detection was found to be 0.2 mg peanuts/kg matrix in both matrices; the linear
response range was demonstrated from 2.5 to 15 mg peanuts/kg matrix by performing statistical tests (homoscedasticity and
Mandel fitting tests). Good accuracy and recovery (>80 ± 2%) were obtained. Different food samples were tested and the results
were compared with those obtained with a commercially available ELISA kit. The results obtained in this work demonstrated
the applicability of the immunomagnetic ELISA methods on real samples and the possibility to perform the assay with significantly
reduced reagent and sample consumption. |
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