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Willem Haasnoot 1 , Ana Frangolho 2 , Gerardo Marchesini 2 , Andrea Valsesia 2 and Claus Nielsen 3

On-site detection of food allergens EU-Marie Curie IAPP: IMPR oved food safety monitoring through E nhanced imaging Nanopla S monic S . IMPRESS (2011-2015). 1 RIKILT P.O. Box 123, 6700 AB Wageningen Contact: willem.haasnoot@wur.nl T + 31 (0)317 48 50 79 www.wageningenur.nl/RIKILT.

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Willem Haasnoot 1 , Ana Frangolho 2 , Gerardo Marchesini 2 , Andrea Valsesia 2 and Claus Nielsen 3

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  1. On-site detection of food allergensEU-Marie Curie IAPP: IMPRoved food safety monitoring through Enhanced imaging NanoplaSmonicS. IMPRESS (2011-2015) 1 RIKILT P.O. Box 123, 6700 AB Wageningen Contact: willem.haasnoot@wur.nl T + 31 (0)317 48 50 79 www.wageningenur.nl/RIKILT Willem Haasnoot1, Ana Frangolho2, Gerardo Marchesini2, Andrea Valsesia2and Claus Nielsen3 2 1 3 Monoclonal antibodies against (total) protein High density peptide array of (allergenic) proteins Imaging Nanoplasmonics biosensor Partners + expertise Optical Label-free 2 mln 15-mer peptides Peanut Hazelnut Egg Milk Soy Prototype “Impressor” Array of synthetic peptides Exchange of knowledge Multiplex immunoassay in food Synthetic peptides Antibodies + protein sequences for epitope mapping Cow milk protein (k-casein) detection using a synthetic peptide A B Improved sensitivity Improved response and stability Results of the first example Figure 1. (A) Absolute responses obtained in the Biacore after the immobilization of k-casein (mw 19000 Da) and the epitope-related peptide (mw 1246 Da) and after the binding of the monoclonal antibody (Mab), showing the higher responses and better stability (during 75 cycles of Mab injection and regeneration) with the immobilized peptide. (B) An improved sensitivity is observed by the application of the immobilized peptide in combination with a lower amount of Mab. Conclusions The exchange of knowledge results in the identification of monoclonal-specific epitopes on food proteins. Biosensor chips with epitope-related synthetic peptides give high stability (reusable) and improved sensitivity. An array of allergenic protein-related peptides with a mixture of the specific monoclonal antibodies in a label-free transportable biosensor will have high (commercial) value for the on-site detection of food allergens. Acknowledgement We would like to thank the European Commission (EU-Marie Curie IAPP programme) and the Dutch Ministry of Economic affairs, Agriculture and Innovation for their financial supports. November 2012, RIKILT Symposium 2 Plasmore SRL, Via G Deledda, Ranco VA, Italy 3 Schafer-N APS,Kystvei 6, Humlebaek, 3050 Denmark

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