Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts

de la Cruz, Silvia and Madrid, Raquel and García, Aina and Alcocer, Marcos and Martín, Rosario and González, Isabel and García, Teresa (2017) Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts. Journal of the Science of Food and Agriculture . ISSN 1097-0010

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Abstract

BACKGROUND

Almonds and Brazil nuts are widely consumed allergenic nuts whose presence must be declared according to food labelling regulations. Their detection in food products has been recently achieved by ELISA methods with recombinant antibodies (scFv) isolated against complete Brazil nut and almond protein extracts. The screening of phage-scFv libraries against complete protein extracts confers a series of advantages over the use of purified proteins, as recombinant proteins might alter their native folding. However, using this strategy, the nature of the target detected by phage-displayed antibodies remains unknown, and requires further research to identify whether they are nut allergens or other molecules present in the extract, but not related to their allergenic potential.

RESULTS

Electrophoretic, chromatographic, immunological and spectrometric techniques revealed that the Brazil nut (BE95) and almond (PD1F6 and PD2C9) specific phage-scFvs detected conformational epitopes of the Brazil nut and almond 11S globulins, recognized by WHO/IUIS as Ber e 2 and Pru du 6 major allergens. Circular dichroism data indicated that severe heat treatment would entail loss of epitope structure, disabling scFv for target detection.

CONCLUSIONS

The presence of important Brazil nut and almond allergens (Ber e 2 and Pru du 6) in foodstuffs can be determined by using phage-display antibodies BE95, PD1F6 and PD2C9 as affinity probes in ELISA.

Item Type: Article
Additional Information: This is the peer reviewed version of the following article: de la Cruz, S., Madrid, R., García, A., Alcocer, M., Martín, R., González, I. and García, T. (), Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts. J. Sci. Food Agric.. Accepted Author Manuscript, which has been published in final form at http://dx.doi.org/10.1002/jsfa.8640. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving
Schools/Departments: University of Nottingham, UK > Faculty of Science > School of Biosciences > Division of Nutritional Sciences
Identification Number: 10.1002/jsfa.8640
Depositing User: Eprints, Support
Date Deposited: 05 Sep 2017 07:27
Last Modified: 05 Sep 2017 23:35
URI: http://eprints.nottingham.ac.uk/id/eprint/45438

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