Molecularly Imprinted Biotin Polymer for Isolation and Detection of Zika-Specific Oligonucleotides Biotinylated Viruses with Lateral Flow and Electrochemical System

Abstract

The Zika virus is a causative agent of infectious diseases that spreads in humans through the mosquitoes Aedes aegypti and Aedes albopictus, which are vectors of the virus. There are several transmission mechanisms, including mother to child transmission during pregnancy, which can lead to microcephaly in newborns and other birth defects. Efforts to monitor and control Zika virus infection globally are very limited and appear to be insufficient due to lack of accountability, lack of organization and unavailability of drugs, vaccines and diagnostic tools. In the context of the development of new diagnostic methodologies, the construction of biological sensors based on the use of oligonucleotide sequences, also known as genosensores, has become a broad activity aiming, mainly, questions of health care and problems with the environment. The development of these genossensores, it looks for the development of specific devices, of low cost, of easy handling and preparation. Biotin is a vitamin that can be coupled to different types of biomolecules due to the high affinity constant with streptavidin. This strong streptavidin-biotin interaction is extremely useful for attaching biotinylated biomolecules to organic or inorganic solid support materials. Biotinylation provides a versatile tool for marking biomolecules in order to obtain an optical reading, fluorescence or electrochemistry, with greater sensitivity. This interaction has been used for years in many bioanalytical methods, including DNA-based methods and immunoassays. Immunoassays are based on the antigen-antibody recognition interaction, generally using the antigen as analyte and the antibody as a recognizer; and these assays may be of different types such as Lateral Flow Assays (LFA). However, the current use of biological materials as supports for anchoring other biological materials such as antibodies, proteins or enzymes, in the bioanalysis generates high costs besides presenting low stability under adverse conditions. In this sense a viable option is the synthesis of materials that can mimic the biological interactions of recognition, which was successfully done by molecularly printed polymers (MIP), which have as one of the main characteristics the high stability under extreme conditions of analysis. Therefore, it is intended to develop in this project of research and internship the systems of selective analysis of LFA with polymers molecularly printed in substitution to the antibodies, that are commonly used in these tests. For this, the MIP for biotin synthesized previously, in the research group of Profª. Maria del Pilar, will be immobilized on the LF tape by different routes (occlusion or covalent methods); and electrochemically using the magnetic MIP for biotin also previously synthesized in the group, the analysis parameters will be optimized and the recognition performance of MIP will be compared with respect to the antibodies. Devices developed after appropriately characterized and with optimized parameters (selectivity, interaction time, etc.) will be applied in the immobilization of biotinylated DNA sequences specific to Zika virus and quantification of their complementary sequence. In this way, it is important to carry out this internship in academic training as a Master's student in Biotechnology, where it will be possible to learn the LFA technique, in addition to the contact with other techniques used in biomolecular analysis, which is the main area of research by Prof. Isabel Pividori.