ERA-NET project 08185 Detection of traces of allergens

ERA-NET project 08185 Detection of traces of allergens in foods (Topic: Allergens) Jorge Martínez Quesada Project Coordinator Dra. Idoia Postigo Resa Department of Immunology, Microbiology and Parasitology. Faculty of Pharmacy. University of the Basque Country. Basqur Country. Spain.

DETECTION OF TRACES OF ALLERGENS IN FOODS Topic: Allergens Project start: march 1 st, 2009. Project end: march 1 st, 2011.

The Parties to this Consortium Agreement Partner 1. (Owner). University of the Basque Country. Department of Immunology, Microbiology and Parasitology. Faculty of Pharmacy. Vitoria/Gasteiz. Araba. Basque Country. Spain. Jorge Martínez Quesada Ph. D. Professor Senior Lecturer (Project Coordinator). Partner 2. VTT Technical Research Centre of Finland. Espoo. Finland. Kristiina Takkinen, Ph. D. Research Professor. Partner 3. Food Research Institut Prague. Department of Nutritive Substances. Prague. Czech Republic. Dana Gabrovska. Ph. D. Head of Department. Partner 4. AZTI – Tecnalia. Unidad de Investigación Alimentaria. Parque Tecnológico. Derio. Bizkaia. Basque Country. Spain. Miguel Ángel Pardo Ph. D. Senior Researcher.

Main objectives According to the background of the topic, the objectives of each member and the potential for common goals, the main objectives were defined as follows: - Production of in-house references to use in ring trials (collaborative studies). - Assessment of the performance of detection methods (ELISA) towards processed allergenic foods: determination of the food processing effect on the extraction and detection of allergens in the selected foodstuff (e. g. milk), i. e. evaluation of the allergens content (decrease/increase) - Isolation of human Ig. E antibodies against food allergens and their use to develop sensitive and specific detection for the allergens in food products and consumer good. - Development of an innovative methodology to obtain novel recombinant antibodies against allergenic proteins (i. e. Anisakis simplex ) which can be used for sensitive allergen detection in food matrix. - Implementation and development of ELISA assays for the sensitive determination of allergens (casein, egg white proteins, β-lactoglobulin, BSA and mustard proteins). Establishment of ring trials (collaborative studies).

ORGANIZATION AND WORK PROGRAM WP 0. Coordi nation (UPV) WP 1. Establishment of homogeneous reference systems (allergens and validated detection methods) (UPV, FRIP, VTT) In house allergen references Commercial ELISA Kits WP 2. Evaluation of food processing on allergen detectability (AZTI) Impact of technology Impact of matrix WP 3. Development of innovative methodologies for allergen traces detection (VTT, AZTI, UPV) Antibodies ELISA kits Immunoarrays WP 4. Validation of detection methods with collaborative studies (FRIP, VTT, UPV, AZTI) Ring Test WP 5. Diffusion and exploitation of results (AZTI, UPV, FRIP, VTT) Finished Ongoing

KEY FACTORS SUPPORTING THE CONCEPT OF EUROPEAN COLLABORATIVE WORK FOR THIS PROJECT AND EXPEDCTED BENEFITS üThe parameters that define (sensitivity, specificity, etc) the current diagnostic methods for the detection of allergens in foods need to be improved. TO IMPROVE THESE PARAMETERS TO OFFER MORE SENSITIVE AND SPECIFIC METHODS TO TRACES ALLERGEN DETECTION ü Validated methods need to be obligatory to ensure that food labelling is in the compliance with legal demands. üVALIDATION ACCORDING TO THE LEGAL REQUIREMENTS üThe development of new validated and homogeneous reagents and standards allows to fulfil more accurately the detection limits required for food allergens. üTo work under the same protocol and with the same validated standards are the key elements for the successful evaluation and comparison of the results obtained within the consortium partners. USE OF WELL KNOWN METHODS WITH WIDE CONSENSUS. INVESTIGATION OF NEW REAGENTS AND NEW NEEDS TO COVER THE MAXIMUM OF ACCURACY

EXPECTED RESULTS AND IMPLEMENTATION ü To set homogeneous reference systems (standards, allergens, antibodies and validated detection methods). ü Evaluation of food processing on allergen detectability ü Development of innovative methodologies and reagents for allergen traces detection ü Validation of detection methods through collaborative studies (ring trials) Diffusion and exploitation of results (Institutions, patents –if required, publications and congresses communications, position reports, authorities, consumers associations, bussiness companies, QC departments, research labs, …)

• How will your project strengthen European food safety? • How will the project results be implemented? • Define three main target groups of your project • How does your project benefit from being a European collaboration project?

1. How will your project strengthen European food safety? The labeling of the content of allergens in foods is a must • Needs: Standardization of methods to measure traces of allergens in foods Ring trials: Multicentric evaluation of the ELISA as basic assay for the detection of traces of allergens in foods (egg, milk, mustard, wheat) • Needs: Development of alternative methods to detection of trace of allergens in foods Development of chimeric antibodies with different sensitivity and specificity to quantify a high number of allergens using multiparametric platforms Use of multiparametric platforms (array) for detecting food allergens in only one test by means of inhibition assays • Needs: Detection of traces of allergens through their encoding genes Detection of allergens of fish parasites (Anisakis) using “omics” alternative methods.

2. How will the project results be implemented? • Availability of validated tests with universal applicability Book of Abstracts: page 508, ISSN 978 -80 -7080 -726 -2 ELISA kit for determination of egg white proteins – Collaborative study Petr Cuhra 1, Petr Hanák 2, Květa Tomková 3, Pavla Dvorská 4, Hana Bulawová 5, Jorge Guisantes 6, Idoia Postigo 7, Marja-Leena Laukkanen 8, Ana Baranda 9, Iñigo Martínez de Marañon 10 Book of Abstracts: page 498, ISSN 978 -80 -7080 -726 -2 ELISA kit for mustard protein determination – Collaborative study Dana Gabrovská 1, Jana Rysová 2, Ingrid Malmheden Yman 3, František Štumr 4, Jan Plicka 5, Stefania Iametti 6, Jorge Martínez 7, Kristiina Takkinen 8, Martin Kubík 9, Migual Angel Pardo 10 • Advances in the detection of non available allergens by DNA and multiparametric tests.

3. Define three main target groups of your project 1. Standardization of classical methods to quantify traces of allergens in foods 2. Development of alternative methods with high sensitivity and specificity 3. To transfer the need of the food labeling and the need of legislation to the interested forum: people, experts and authorities International Innovation Annual Research Report

4. How does your project benefit from being a European collaboration project? • Developing and validation of new tests and new technologies for the detection of allergens in foods. • Unification of valid criteria for reporting on the composition and quantity of relevant food allergens. • Agreements for the labeling. • Clinically relevant allergen panel design.

Thank you very much