RT Journal Article
T1 Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products
A1 Cruz, Silvia de la
A1 Cubillos-Zapata, Carolina
A1 López-Calleja, Inés María
A1 Ghosh, Satyabrata
A1 Alcocer, Marcos
A1 González Alonso, María Isabel
A1 Martín De Santos, María Del Rosario
A1 García Lacarra, Teresa
AB 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 (110–120 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.
PB ELSEVIER
SN 0956-7135
YR 2015
FD 2015
LK https://hdl.handle.net/20.500.14352/94191
UL https://hdl.handle.net/20.500.14352/94191
LA eng
NO de la Cruz, S., Cubillos-Zapata, C., López-Calleja, I. M., Ghosh, S., Alcocer, M., González, I., Martín, R., & García, T. (2015). Isolation of recombinant antibody fragments (scFv) by phage display technology for detection of almond allergens in food products. Food Control, 54, 322-330. https://doi.org/10.1016/J.FOODCONT.2015.02.011
NO Comunidad de Madrid
NO Ministerio de Economía y Competitividad (España)
DS Docta Complutense
RD 7 abr 2025