Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products

Cruz, Silvia de la and Cubillos Zapata, Carolina and López Calleja, Inés María and Ghosh, Satyabrata and Alcocer, Marcos and González García, Isabel and Martín, Rosario and García Lacarra, Teresa (2015). Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products. "Food Control", v. 54 ; pp. 322-330. ISSN 0956-7135. https://doi.org/10.1016/j.foodcont.2015.02.011.

Description

Title: Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products
Author/s:
  • Cruz, Silvia de la
  • Cubillos Zapata, Carolina
  • López Calleja, Inés María
  • Ghosh, Satyabrata
  • Alcocer, Marcos
  • González García, Isabel
  • Martín, Rosario
  • García Lacarra, Teresa
Item Type: Article
Título de Revista/Publicación: Food Control
Date: August 2015
ISSN: 0956-7135
Volume: 54
Subjects:
Faculty: E.T.S.I. Agrónomos (UPM) [antigua denominación]
Department: Ingeniería Agroforestal
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Tree nut allergies are considered an important health issue in developed countries. To comply with the regulations on food labeling, reliable allergen detection methods are required. In this work we isolated almond-specific recombinant antibody fragments (scFv) from a commercial phage display library bypassing the use of live animals, hence being consistent with the latest policies on animal welfare. To this end an iterative selection procedure employing the Tomlinson I phage display library and a crude almond protein extract was carried out. Two different almond-specific scFv (named PD1F6 and PD2C9) were isolated after two rounds of biopanning, and an indirect phage ELISA was implemented to detect the presence of almond protein in foodstuffs. The isolated scFvs demonstrated to be highly specific and allowed detection of 40 ng mL?1 and 100 ng mL?1 of raw and roasted almond protein, respectively. The practical detection limit of the assay in almond spiked food products was 0.1 mg g?1 (110e120 ppm). The developed indirect phage ELISA was validated by analysis of 92 commercial food products, showing good correlation with the results obtained by a previously developed real-time PCR method for the detection of almond in foodstuffs. The selected phage clones can be affinity maturated to improve their sensitivity and genetically engineered to be employed in different assay formats.

Funding Projects

TypeCodeAcronymLeaderTitle
Government of SpainAGL2010-15279UnspecifiedUnspecifiedDetección de frutos secos alergénicos en alimentos mediante péptidos y anticuerpos recombinantes (ELISA y microarrays) y sondas genéticas (PCR múltiple en tiempo real)
Madrid Regional Government2009/AGR-1489UnspecifiedUnspecifiedUnspecified
Government of SpainAP2010-0028UnspecifiedUnspecifiedUnspecified

More information

Item ID: 41191
DC Identifier: http://oa.upm.es/41191/
OAI Identifier: oai:oa.upm.es:41191
DOI: 10.1016/j.foodcont.2015.02.011
Official URL: http://www.sciencedirect.com/science/article/pii/S0956713515000924
Deposited by: Memoria Investigacion
Deposited on: 23 Jun 2016 15:35
Last Modified: 06 Jun 2019 16:12
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