Person:
García Lacarra, Teresa

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First Name
Teresa
Last Name
García Lacarra
URI
https://hdl.handle.net/20.500.14352/80401
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Nutrición y Ciencia de los Alimentos
Area
Nutrición y Bromatología
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Author: Martín De Santos, María Del Rosario × Subject: 61 ×

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Now showing 1 - 3 of 3
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    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, 2013) Cruz, Silvia de la; López-Calleja, Inés María; Alcocer, Marcos; González Alonso, María Isabel; Martín De Santos, María Del Rosario; García Lacarra, Teresa
    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.
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    High resolution TaqMan real-time PCR approach to detect hazelnut DNA encoding for ITS rDNA in foods
    (Food Chemistry, 2013) López-Calleja, Inés María; Cruz, Silvia de la; Pegels, Nicolette; González Alonso, María Isabel; García Lacarra, Teresa; Martín De Santos, María Del Rosario
    A broad range of foods have been described as causing allergies, but the majority of allergic reactions can be ascribed to a limited number of food components. Recent extensive surveys showed how tree nuts, particularly hazelnut (Corylus avellana L.) seeds, rank amongst the most important sources of food allergy. In order to protect the allergic consumer, efficient and reliable methods are required for the detection of allergenic ingredients. For this purpose, we have developed a real-time polymerase chain reaction (PCR) for detection of hazelnut in commercial food products. In this way a specific hazelnut primer pair based on the ITS marker (70 bp) and a nuclease (TaqMan) probe labelled with FAM and BHQ were designed. Sensibility of real-time PCR was determined by analysis of raw and heat treated hazelnut-wheat flour mixtures with a range of detection of 0.1–100,000 ppm. Practical applicability of the real-time PCR assay developed for determining hazelnut in different food matrices was investigated by analyzing 179 commercial foodstuffs comprising snacks, biscuits, chocolates, bonbons, creams, nut bars, ice creams, precooked meals, breads, beverages, yogurts, cereals, meat products, rice cake and nougat. From the total of samples analyzed, 40 commercial food products that didn’t declare hazelnut nor traces on the label were found to contain hazelnut. The real-time PCR method proposed herein due to its high sensitivity facilitates the detection of hazelnut traces in commercial food products and can also be useful for monitoring the effectiveness of cleaning processes and as consequence, can help to prevent the food allergic consumer from unintentional ingestion of hidden allergens.
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    TaqMan real-time PCR assay for detection of traces of Brazil nut (Bertholletia excelsa) in food products
    (Food Control, 2013) Cruz, Silvia de la; López-Calleja, Inés María; Alcocer, Marcos; González Alonso, María Isabel; Martín De Santos, María Del Rosario; García Lacarra, Teresa
    Mislabeling of food products containing Brazil nut may represent a serious thread for allergic consumers. In order to protect sensitized individuals, reliable methods to detect trace amounts of Brazil nut must be accessible to food industry as well as Food Safety authorities. According to this, a TaqMan real-time polymerase chain reaction (PCR) method has been developed for specific detection of Brazil nut in foodstuffs. The method employs Brazil nut specific primers and probe, targeting 2S albumin gene (Ber e 1), and a positive amplification control based on 18S rRNA gene. Results obtained on sensitivity with wheat flour spiked with different concentrations of raw and heat treated Brazil nut showed that the limit of detection (LOD) for the technique was 2.5 mg/kg. Applicability of the Brazil nut specific system was assessed through analysis of 66 different commercial food samples. The reported real-time PCR assay provides a useful tool for detection of Brazil nut DNA, and it can be used as a routine analysis to assert accuracy on food labeling.