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

• Grant number: 16/01308-7
• Support type: Scholarships in Brazil - Doctorate
• Effective date (Start): October 01, 2016
• Effective date (End): January 31, 2019
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Science
• Principal researcher: João Roberto Oliveira do Nascimento
• Grantee: Gabriela Justamante Handel Schmitz
• Home Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 13/07914-8 - FoRC - Food Research Center, AP.CEPID
• Associated scholarship(s): 16/22671-2 - Cloning, recombinant expression and validation by microarray of potentially allergenic food proteins, BE.EP.DR

Abstract

In some groups of food, the allergy to a particular allergen of the group can involve an allergic response to other agents 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. Some foods that are object of interest in the clinical area in Brazil are pineapple, papaya, mango and cassava. The paucity of identified allergens to these organisms prevents the diagnosis of food allergy, thus, it is important expanding the allergens platform by obtaining new allergenic proteins by recombinant DNA technology for use in microarrays. Therefore, the aim of this work is cloning, expressing and purifying potentially allergenic proteins of important foods from Brazil. These proteins will be used in microarray production and validation by assays with the serums from allergic patients to selected foods. Achieving this goal, the potentially allergenic proteins of these organisms will be studied, which will be identified by taxonomic similarity and by 2D Western Blot technique coupled mass spectrometry. From the in silico analysis, primers consensus will be designed for amplification of the cDNA sequences of potential allergens, whereas for the 2D Western Blot will be used serums from allergic patients to selected foods. Microarray validation will allow the acceleration of the diagnosis, indicating the presence of sensitization for allergens of cross-reactivity.