Dra. Idoia Postigo ResaDepartment of Immunology, Microbiology and Parasitology.

Faculty of Pharmacy. University of the Basque Country.

Basqur Country. Spain.

ERA-NET project 08185

Detection of traces of allergens in foods(Topic: Allergens)

Jorge Martínez QuesadaProject Coordinator

DETECTION OF TRACES OF ALLERGENS IN FOODS

Project start: march 1st, 2009.

Project end: march 1st, 2011.

Topic: Allergens

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.

The Parties to this Consortium Agreement

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 IgE 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).

Main objectives

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According to the the background of the topic, the objectives of each member and the potential for common goals, the main objectives were defined as follows:

WP 1. Establishment of homogeneous reference systems (allergens and validated detection methods)

(UPV, FRIP, VTT)In house allergen references

Commercial ELISA Kits

WP 3. Development of innovative

methodologies for allergen traces

detection(VTT, AZTI, UPV)

AntibodiesELISA kits

Immunoarrays

WP 4. Validation of detection methods with collaborative studies (FRIP, VTT, UPV, AZTI)

Ring Test

WP 2. Evaluation of food processing on

allergen detectability

(AZTI)Impact of technology

Impact of matrix

WP 0.

Coordination (UPV)

WP 5. Diffusion and exploitation of results(AZTI, UPV, FRIP, VTT)

ORGANIZATION AND WORK PROGRAM

Finished

Ongoing

The 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

Validated methods need to be obligatory to ensure that food labelling is in the compliance with legal demands.

VALIDATION ACCORDING TO THE LEGAL REQUIREMENTS

The development of new validated and homogeneous reagents and standards allows to fulfil more accurately the detection limits required for food allergens. To 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

KEY FACTORS SUPPORTING THE CONCEPT OF EUROPEAN COLLABORATIVE WORK FOR THIS PROJECT AND EXPEDCTED BENEFITS

To set homogeneous reference systems (standards, allergens, antibodies and validated detection methods).

Evaluation of food processing on allergen detectability

Development of innovative methodologies and reagents for allergen traces detection

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, …)

EXPECTED RESULTS AND IMPLEMENTATION

• How will your project strengthen European food safety?

• How will the project results be implemented?

• Define the 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 studyPetr Cuhra1, Petr Hanák2, Květa Tomková3, Pavla Dvorská4, Hana Bulawová5, Jorge Guisantes6, Idoia Postigo7, Marja-Leena Laukkanen8,Ana

Baranda9, Iñigo Martínez de Marañon10

Book of Abstracts: page 498, ISSN 978-80-7080-726-2

ELISA kit for mustard protein determination – Collaborative studyDana Gabrovská1, Jana Rysová2, Ingrid Malmheden Yman3, František Štumr 4, Jan Plicka5, Stefania Iametti6, Jorge Martínez7, Kristiina Takkinen8, Martin Kubík9, Migual Angel Pardo10

• Advances in the detection of non available allergens by DNA and multiparametric tests.

3. Define the 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