Method validation for the extraction and quantification of proteins in genetically modified plants and allergenic proteins present in foods by the Enzyme Linked ImmunonoSorbent Assay (ELISA)

Abstract

With the advent of genetically modified foods to the market, there was a need to carry out a safety assessment of organisms derived from biotechnology in relation to human and animal health as well as the environment. Internationally the principle of substantial equivalence has been adopted in the safety assessment of GMOs, which consists of comparing the characteristics of the modified plant with the conventional plant. The National Technical Biosafety Commission (CTNBio), responsible for the approval of transgenic foods in Brazil, also adopts the principle of substantial equivalence for the evaluation of biosafety of GMOs based on Law 11,105 / 2005 and Normative Resolution No. 5 of 2008.Since 2006, Tecam has performed studies on the composition of GMOs that involves macro, micronutrients, anti-nutrients and metabolites, but does not quantify the transgenic proteins expressed in the plant. The companies that produce the GMOs carry out these analyzes in the laboratories of their research centers or laboratories providing service abroad due to the lack of training of laboratories that provide service in this specialty in the country. The project aims to develop this know-how, which today is restricted to the national academic area and external laboratories, to increase the competitiveness of the company. The laboratory structure and differentiated technique required for protein analysis, which began in Phase I and will be finalized in Phase II, will allow the opening of a new area of activity and knowledge in the company. By offering a more complete portfolio of services to its clients, the company adds value per sample received as well as being able to conquer new clients in the area of biotechnology. The laboratory of expression and quantification of protein will also enable the growth of the company in other segments, today the food market is obliged to inform the consumer the presence of allergenic proteins in their products. This technological development in the company also serves the interests of the country because the services contracted abroad can be contracted in the domestic market. In phase I, we performed a pilot experiment in which a standard curve was established for the 2m-EPSPS protein (the enzyme 5-enolpyruvylhikimate-3-phosphate synthase) and demonstrated the ability to extract and quantify the 2m-EPSPS protein in lyophilized samples of Cotton fortified with said protein. In phase II we propose to determine the content of the protein 2mEPSPS in different tissues and in the different phases of the development of the cotton in natura. The challenge of this phase is to deploy a robust procedure that is sensitive enough to discriminate different amounts of modified protein according to the expression of the gene in the different parts and stages of development of the plant. In Phase III the new area of operation of the company will allow the implementation of procedures for the analysis of protein from other vegetable crops according to market interest. In particular, we will be able to serve national companies such as EMBRAPA that develops GM plants mainly destined to the domestic market, in the case of the GM beans that it has developed. (AU)