Cloning, recombinant expression and validation by microarray of potentially allergenic food proteins

• Grant number: 16/22671-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: February 01, 2017
• End date: January 31, 2018
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Science
• Principal Investigator: João Roberto Oliveira do Nascimento
• Grantee: Gabriela Justamante Handel Schmitz
• Supervisor: Marcos JC Alcocer
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Nottingham, Sutton Bonington, England
• Associated to the scholarship: 16/01308-7 - Cloning, recombinant expression and validation by microarray of potentially allergenic food proteins., BP.DR

Abstract

Food allergy is an adverse effect to human health and it occurs due to a specific immune response triggered for an allergen, which occurs reproducibly on exposure to a particular food. In some groups of food, the allergy to a particular allergen of the group can trigger an allergic response to other foods from the same group and this phenomenon is known as cross-reactivity. A small and specific group of protein families presents in food is responsible for a large number of cases of allergic reactions and the majority of allergic reactions are mediated by IgE, which can be detected by microarray. This miniaturized allergy test allows the simultaneous detection of a large number of allergens and it provides the acceleration of diagnosis and the discovery of cross-reactivity. Therefore, there is a great interest in identification of potential food allergens and their achievement by recombinant DNA technology for application in microarrays. Recombinant allergens require correct folding to form conformational epitopes and yeasts have several advantages in relation to this requirement in comparison to traditional bacteria expression system, for example, E.coli. Pichia pastoris has develop great interest among the available yeast systems because it is more efficient in producing large quantities of properly folded, soluble and biologically active allergens. Four Brazilian foods, cassava, mango, papaya and pineapple are object of interest in the clinical area in Brazil and it is necessary a further evaluation about their potential allergens, since there is a limited information about them. Therefore, this project will aim the recombinant expression of selected potential allergenic proteins regarding these Brazilian foods in Pichia pastoris and their use in microarrays production towards diagnosis. (AU)