RT Journal Article
T1 Selection of Recombinant Antibodies by Phage Display Technology and Application for Detection of Allergenic Brazil Nut (Bertholletia excelsa) in Processed Foods
A1 Cruz, Silvia de la
A1 López-Calleja, Inés María
A1 Alcocer, Marcos
A1 González Alonso, María Isabel
A1 Martín De Santos, María Del Rosario
A1 García Lacarra, Teresa
AB Current immunological methods for detection of Brazil nut allergens in foods are based on polyclonal antibodies raised in animals. Phage display technology allows the procurement of high-affinity antibodies avoiding animal immunization steps and therefore attaining the principle of replacement supported by animal welfare guidelines. In this study, we screened Tomlinson I and J libraries for specific binders against Brazil nut by employing a Brazil nut protein extract and a purified Brazil nut 2S globulin, and we successfully isolated a phage single chain variable fragment (named BE95) that specifically recognizes Brazil nut proteins. The selected phage scFv was further used as affinity probe to develop an indirect phage-ELISA for detection of Brazil nut in experimental binary mixtures and in commercial food products, with a limit of detection of 5 mg g(-1). This study describes for the first time the isolation of recombinant antibody fragments specific for an allergenic tree nut protein from a naïve library and paves the way to develop new immunoassays for food analysis based on probes that can be produced in vitro when required and do not rely on animal immunization.
PB ACS Publications
SN 0021-8561
YR 2013
FD 2013
LK https://hdl.handle.net/20.500.14352/94172
UL https://hdl.handle.net/20.500.14352/94172
LA eng
NO de la Cruz, S., López-Calleja, I. M., Alcocer, M., González, I., Martín, R., & García, T. (2013). Selection of recombinant antibodies by phage display technology and application for detection of allergenic Brazil nut (Bertholletia excelsa) in processed foods. Journal of agricultural and food chemistry, 61(43), 10310–10319. https://doi.org/10.1021/jf403347t
NO Ministerio de Economía y Competitividad (España)
NO Comunidad de Madrid
DS Docta Complutense
RD 1 sept 2024