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

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...

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Main Authors: de la Cruz, Silvia, Madrid, Raquel, García, Aina, Alcocer, Marcos, Martín, Rosario, González, Isabel, García, Teresa
Format: Article
Published: Wiley 2017
Online Access:https://eprints.nottingham.ac.uk/45438/
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author de la Cruz, Silvia
Madrid, Raquel
García, Aina
Alcocer, Marcos
Martín, Rosario
González, Isabel
García, Teresa
author_facet de la Cruz, Silvia
Madrid, Raquel
García, Aina
Alcocer, Marcos
Martín, Rosario
González, Isabel
García, Teresa
author_sort de la Cruz, Silvia
building Nottingham Research Data Repository
collection Online Access
description 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.
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spelling nottingham-454382020-05-04T19:02:39Z https://eprints.nottingham.ac.uk/45438/ 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 Madrid, Raquel García, Aina Alcocer, Marcos Martín, Rosario González, Isabel García, Teresa 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. Wiley 2017-08-28 Article PeerReviewed de la Cruz, Silvia, Madrid, Raquel, García, Aina, Alcocer, Marcos, Martín, Rosario, 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, 30 (6). pp. 779-790. ISSN 1097-0010 http://onlinelibrary.wiley.com/doi/10.1002/jsfa.8640/abstract doi:10.1002/jsfa.8640 doi:10.1002/jsfa.8640
spellingShingle de la Cruz, Silvia
Madrid, Raquel
García, Aina
Alcocer, Marcos
Martín, Rosario
González, Isabel
García, Teresa
Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title_full Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title_fullStr Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title_full_unstemmed Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title_short Identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scFv) obtained against Brazil nut and almond extracts
title_sort identification and characterization of the proteins bound by specific phage-displayed recombinant antibodies (scfv) obtained against brazil nut and almond extracts
url https://eprints.nottingham.ac.uk/45438/
https://eprints.nottingham.ac.uk/45438/
https://eprints.nottingham.ac.uk/45438/