Food Allergen Methods – Development of a coordinated research programme for food allergens – identification, modification and in vivo responses

The Norwegian Food Safety Authority (NFSA) tasked the Norwegian Veterinary Institute (NVI) with the development of methods for food allergen detection in 1999. A working group on food allergens, established at NVI under the leadership of Eliann Egaas, started the evaluation of commercial ELISA kits, conducted food surveys in commission of NSFA and produced antibodies for the development of in-house ELISAs.

Survey results were published in official reports:

  • 2001: Survey of casein in baby food, meat products, cookies, snacks, sorbets and sweets
  • 2003: Survey of casein in food products on the Norwegian marked
  • 2006: Survey of peanuts in food products labelled «may contain traces of peanut»
  • 2006: Survey of hazelnuts in food products labelled «may contain traces of hazelnut»

The establishment of a competence center for food allergen analytics at NVI was further supported by research grants from the Norwegian Research Council.

  • 1999-2003: Allergens in food on the Norwegian marked
  • 2003–2008: Development of a co-ordinated research programme for food allergens – identification, quantification, modification and in vivo responses (NFR153436).

It was a major aim of the project to develop a method portfolio for the most important allergens in food that can cause IgE-mediated allergic incidents. The EU has identified 14 foodstuffs that have to be declared on all prepacked products, and recently also in restaurants and over-the-counter meals. In addition, methods for gluten detection, critical for the survey of gluten-free claims, were developed with the aim to protect consumers with gluten intolerance (coeliac disease).

The food allergen detection methods were validated in accordance to international standards. Quality was assured through participation in international ring trials and knowledge exchange with experts in international method standardisation bodies such as CEN (TC275WG12 Food Allergens) and AOAC (Food Allergen Community). These cooperations are continued to date. Furthermore, NVI is the national reference laboratory for food allergens and participating in the EU-initiative ENFADL (European Network of Food Allergen Detection Laboratories).    

The Food Allergen Group at NVI was responsible for the identification of the eliciting allergens in food samples submitted to the Norwegian National Register of Severe Allergic Reactions to Food (Matallergiregisteret – Meldesystem og register for alvorlige allergiske reaksjoner på mat) (2000-2018). The food allergen register was administered by the Norwegian Institute of Public Health (NIPH) in collaboration with the NFSA and NVI. Severe allergy cases were defined as reactions that lead to the patient contacting a doctor within 24 hours. Doctors sent a case report and a serum sample for specified IgE analysis to NIPH, whereas food samples were analysed and NVI.

Two novel food allergens in Norway, lupine in bakery goods and fenugreek in Indian-style food products, were discovered through this cooperation. Lupine was later included in the EU-list of 14 important foodstuffs.   

The portfolio at NVI offered analytical method for all relevant food allergens. Since the project end in 2008, numerous new commercial ELISA kits have become available. In addition, PCR methods are used for some allergens, and LC-MS/MS methods have been developed for the most important allergens.

 Allergen Principle Source LOD
Peanut ELISA commercial kit 0.5 mg/kg
Egg ELISA commercial kit 0.5 mg/kg
Fish ELISA in-house > commercialised 0.5-1 mg/kg
Fish (multi) Immunoblot in-house qualitatively
Milk RIE in-house 2.5 mg/kg
Milk ELISA in-house 1.0 mg/kg
Hazelnut FIA in-house 0.8 mg/kg
Almond Blot in-house qualitatively
Gluten ELISA commercial kit 20 mg/kg
Crustacean ELISA in-house 1.0 mg/kg
Soya ELISA commercial kit 0.1 %
Lupine ELISA in-house > commercialised 1.0 mg/kg
Sesame ELISA commercial kit 1.0 mg/kg
Mustard Blot in-house qualitatively
Cellery ELISA in-house qualitatively
Fenugreek ELISA in-house qualitatively
Anisakis simplex ELISA in-house 1.1 mg/kg

The project work has also resulted in a number of scientific publications:

Publications

  • Fæste CK, Wiker HG, Løvik M, Egaas E (2003). Hidden shellfish in a fish cake. Allergy 58, 1204-1205.
  • Fæste CK, Løvik M, Wiker HG, Egaas E (2004). A case of peanut cross-allergy to hidden lupine flour in a hot dog bread. Int Arch Allergy Immunol 135, 135-39.
  • Holden L., Fæste CK, Egaas E. (2005) A quantitative sandwich ELISA for the determination of Lupine (Lupinus L.) in Foods. J Agr Food Chem. 53, 5866-5871.
  • Fæste CK, Holden L, Plassen C, Almli B (2006). Sensitive time-resolved fluoroimmunoassays for the detection of hazelnut (Corylus avellana) protein traces in food matrices. J Immunol Methods 314, 114-122.
  • Dooper MMBW, Holden L, Fæste CK, Thompson KM, Egaas E (2007). Monoclonal antibodies against the candidate lupin allergens a-conglutin and b-conglutin. Int Arch Allergy Immunol 143, 49-58.
  • Fæste CK, Løvberg KE, Lindvik H, Egaas E. (2007). Extractability, stability, and allergenicity of egg-white proteins in differently heat-processed foods. J AOAC Int. 90, 427-436.
  • Werner MT, Fæste CK, Egaas E (2007). A quantitative sandwich ELISA for the determination of tromoyosin from crustaceans in foods. J Agr Food Chem 55, 8025-32.
  • Fæste CK, Plassen, C (2008).  Quantitative Sandwich ELISA for the Determination of Fish in Foods. J Immunol Methods 329, 45-55.
  • Holden L, Sletten GBG, Lindvik H, Fæste CK, Dooper MMBW (2008). Characterization of IgE binding to lupin, peanut and almond with sera from lupin-allergic patients. Int Arch Allergy Immunol, 146:267-276.
  • Sletten GBG, Holden L, Egaas E, Fæste CK (2008). Differential influence of the degree of processing on immunogenicity following proteolysis of casein and β-lactoglobulin. Food Agri Immunol 19:213-228.
  • Fæste CK, Namork E, Lindvik H (2009).  Allergenicity and antigenicity of fenugreek proteins (Trigonella foenum-graecum) in foods. J. Allergy Clin Immunol, 123:187-94.
  • Dooper MMBW, Plassen C, Holden L, Lindvik H, Faeste CK (2009). IgE cross-reactivity between lupin conglutins and peanut allergens in serum of lupin-allergic individuals. J Investig Allergol Clin Immunol, 19:283-291.
  • Fæste CK, Jonscher KR, Sit L, Klawitter J, Løvberg KE, Moen LH (2010). Differentiating cross-reacting allergens in the immunological analysis of celery (Apium graveolens) by mass spectrometry. J. AOAC Int. 93:451-461.
  • Fæste CK, Christians U, Egaas E, Jonscher KR (2010). Characterization of potential allergens in fenugreek (Trigonella foenum-graecum) using patient sera and MS-based proteomic analysis. J. Proteomics 73:1321-1333.
  • Fæste CK, Namork E (2010). Differentiated patterns of legume sensitisation in peanut-allergic patients. Food Analyt Meth 3:357-362.
  • Werner MT, Fæste CK, Levsen A, Egaas E (2011). A quantitative Sandwich ELISA for the detection of Anisakis simplex protein in seafood. Eur. Food Res. Technol. 232:157-166.

Book chapters

  • Fæste CK: Lupin allergen detection, Chapter 22, Molecular Biological and Immunological Techniques and Applications for Food Chemists, edt. Popping B, Diaz-Amigo C, Hoenicke K.  John Wiley & Sons, Ltd, 2009.
  • Fæste CK: Allergens in Peanut, Soybean, and Lupin, Chapter 3, Food Allergens: Analysis Instrumentation and Methods, edt. Nollet L. Van Hengel A. Taylor & Francis Group, LLC, Boca Raton, Fl. USA, 2011.

Posters

  • Ferm M, Eriksson A, Malmheden Yman I, Haugland Moen L, Fæste CK. Detection of crustaceans with an antiserum to tropomyosin. VIIth International Conference on Agri-Food Antibodies. 10.-13. September, 2003, Uppsala, Sweden.
  • Holden L, Sletten G, Scarpeid H-J, Fæste CK. Peptic digestion patterns and antigenicity of commercial milk types in Norway. 9th International Symposium on Immunological, Chemical and Clinical Problems of Food Allergy. 18.-21. April 2004, Budapest, Hungary.
  • Fæste CK, Namork E, Omholt-Jensen G, Løvik M, Egaas E. Lupine protein as food allergen. 20.-23. Juni 2004, 8th Nordic Nutrition Conference, Tønsberg, Norway.
  • Fæste CK, Namork E, Omholt-Jensen G, Løvik M, Egaas E. The problem of “hidden” allergens in processed food. 20.-23. Juni 2004, 8th Nordic Nutrition Conference, Tønsberg, Norway.
  • Vinje NE, Egaas E, Fæste CK, Løvik M. Mouse model for food allergy. 20.-23. Juni 2004, 8th Nordic Nutrition Conference, Tønsberg, Norway.
  • Almli B, Omholt-Jensen G, Fæste CK, Egaas E. “May contain traces of nut” – an assurance for the food industry? 20.-23. Juni 2004, 8th Nordic Nutrition Conference, Tønsberg, Norway.
  • Holden, L, Sletten G, Scarpeid H-J, Løvberg K, Fæste CK. Peptic digestion patterns and antigenicity of commercial milk types in Norway. 20.-23. Juni 2004, 8th Nordic Nutrition Conference, Tønsberg, Norway.
  • Dooper MMBW, Holden L, Fæste CK, Thompson KW, Egaas E. The specificity of monoclonal antibodies against storage seed proteins from Lupinus albus. VIII International conference on agri-food antibodies, 6.-9. September 2005, Chester, UK.
  • Dooper MMBW, Holden L, Fæste CK, Thompson KW, Egaas E. Detection of the major parvalbumin by using monoclonal antibodies. VIII International conference on agri-food antibodies, 6.-9. September 2005, Chester, UK.
  • Fæste CK, Løvberg KE, Skarpeid HJ, Egaas E. Performance test of two commercial ELISA-kits for the detection of egg in foods. VIII International conference on agri-food antibodies, 6.-9. September 2005, Chester, UK.
  • Fæste CK, Plassen C, Almli B. Sensitivity tuning in corylin detection – beneficial or hampering? VIII International conference on agri-food antibodies, 6.-9. September 2005, Chester, UK.
  • Fæste CK, Løvberg K, Skarpeid HJ, Egaas E. Comparison of two commercial ELISA-kits for the detection of egg in processed foods. 119th AOAC Annual Meeting and Exposition, 11.-15. September 2005, Orlando, USA.
  • Fæste CK, Holden L, Reisdorph N, Halvorsen R, Løvik M. Lupine food allergy in Norway. 119th AOAC Annual Meeting and Exposition, 11.-15. September 2005, Orlando, USA.
  • Holden L, Fæste CK, Dooper MMBW, Moen LH, Namork E. Occurrence of lupin-containing foods and lupin food allergy in Norway. XXV EAACI, 10.-14. Juni 2006, Wien, Austria.
  • Fæste CK, Namork E. Fenugreek proteins are potential food allergens. 2nd International Symposium on Molecular Allergology, 22.-24. April 2007, Roma, Italy.
  • Fæste CK, Plassen C, Moen LH. Cross-reactivity of anti-cod parvalbumin antibodies with different fish species. 2nd International Symposium on Molecular Allergology, 22.-24. April 2007, Roma, Italy.
  • Løvberg KE, Holden L, Fæste CK. Extraction of egg, lupin and sesame proteins from food. 9th International Symposium on Advances in Extraction Technologies, 3.-6. Juni 2007, Ålesund, Norway.
  • Dooper M, Holden L, Thompson K, Fæste CK. A homologous sequence between Lupinus spp. (lupin) and Prunus dulcis (almond) allergens recognized by an anti-lupin monoclonal antibody. XXVI EAACI, 9.-13. Juni 2007, Göteborg, Sweden.
  • Namork E, Fæste CK, Stensby B, Søyland A, Lindvik H. Fenugreek – a new hidden allergen? XXVI EAACI, 9.-13. Juni 2007, Göteborg, Sweden.
  • Løvberg KE, Fæste CK. Survey of sesame protein foods on the Norwegian marked using a commercial ELISA kit. IXth International Conference on AgriFood Antibodies (ICAFA), 10.-13. September 2007, Vettre, Norway.
  • Plassen C, Jonscher KR, Fæste CK. Might fish troponin C be allergenic? IXth International Conference on AgriFood Antibodies (ICAFA), 10.-13. September 2007, Vettre, Norway.
  • Fæste CK, Jonscher KR, Namork E. Fenugreek – a potential danger for peanut-allergic consumers. 121st AOAC Annual Meeting and Exposition, 16.-20. September 2005, Anaheim, USA.
  • Fæste CK, Namork E. Differentiated patterns of legume sensitisation in peanut-allergic patients. 5th Workshop on Food Allergens, 11.-14. Mai 2008, Halifax, Canada.
  • Løvberg KE, Fæste CK. Traceability of sesame protein in sesame oil. 10th International Symposium on Immunological, Chemical and Clinical Problems of Food Allergy. 26.-29. Mai 2008, Parma, Italy.
  • Namork E, Fæste C, Stensby B, Søyland A, Lindvik H, Løvik M. Fenugreek – a hidden food allergen. ITCASS, 2008 22-23 Sept., Oslo, Norway.
  • Namork E, Fæste CK, Stensby B, Egaas E, Løvik M. Allergic reactions to food in Norway. New Trends in Allergy VII. 22.-24.7. 2010. Munich, Germany
  • Werner MT, Fæste CK, Levsen A, Egaas E. ELISA for the determination of the herring worm Anisakis simplex in foods. 124th AOAC Annual Meeting and Exposition. 26.9.-29.9. 2010. Orlando, FL, USA.

Presentations

  • Fæste CK. Quantitative Sandwich ELISA for the Determination of Fish in Foods. 4th Workshop on Food Allergens, 15.-18. Oktober 2006. Québec, Canada.
  • Fæste CK. Confirmation of a new allergen on the Norwegian market. Seminar on Threshold Levels and Methods in Food Allergy.  16. Oktober 2008. Oslo.
  • Fæste CK. Chasing new food allergens – sources, techniques, strategies. Biologically Active Compounds in Food. International Conference. 15.-16.10.2015, Lodz, Poland.
  • Fæste CK. Assessing the allergenicity risk of novel food (protein)s – current situation. EU COST-Action ImpARAS Stakeholder Meeting. 22.2.2018, Brussels, Belgium.
  • Fæste CK. Identifisering og kvantifisering av allergener i mat. Teknologisk Matforum, Allergener i fokus. 24.10.2018, Oslo, Norway.
  • Fæste CK. Current status of food allergens analysis by LC-MS/MS. 17th meeting of CEN/TC 275/WG 12 “Food allergens”. 14.11.2018, Braunschweig, Germany