| Grant number: | 16/02873-0 |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| Start date: | June 01, 2016 |
| End date: | March 31, 2018 |
| Field of knowledge: | Physical Sciences and Mathematics - Chemistry |
| Principal Investigator: | Carmen Lúcia Cardoso |
| Grantee: | Adriana Ferreira Lopes Vilela |
| Host Institution: | Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil |
| Associated research grant: | 13/01710-1 - Enzyme ligand: new models of screening, AP.TEM |
| Associated scholarship(s): | 16/15037-5 - Co-immobilization of enzymes acetylcholinesterase and beta-secretase 1: study of conditions for screening of ligands, BE.EP.DR |
Abstract The identification of compounds which specifically bind the biological target of great importance in research for development of new drugs assisting in reducing the time and number of candidates. Efficient strategies have been developed and a very useful tool is the bioaffinity chromatography involves the immobilization of a target. This bonding agent may be a sequence of DNA, RNA, antibodies, receptors, membranes, enzymes, proteins, biomimetic dyes, enzyme substrate or inhibitor or any other target molecule. The immobilization of enzymes has shown several advantages compared to online enzymatic activity assays in solution, among them studies, increased stability in the presence of organic solvents without significant loss of catalytic activity, reuse and the use of small quantities of enzyme. If there is more than one target of interest, co-immobilization of enzymes may also be applied. This is a technique that involves the immobilization of two or more targets on the same support and these used simultaneously for screening ligands. In this case, have to create a specific environment that mimics a cell can be considered as models of compartmentalized intracellular enzymes and in vitro models for cellular processes. The use of enzymes immobilized enzyme bioreactors having as stationary phase in chromatographic systems connected high efficiency allows the joining of both characteristics such as selectivity, speed, reproducibility and non destructive testing the specificity and sensitivity of an enzymatic reaction. Enzymes, cholinesterase and secretases, are targets of interest in studies of drug development for Alzheimer's disease, because they are involved in the etiology and progression of the disease. Considering the interest in inhibitors of these enzymes, which act in the presence of both the goal of this project is the development of two methods: (1) involving acetylcholinesterase (AChE) enzyme and b-secretase-1 (BACE1) co-immobilized by entrapment, and (2) immobilization of BACE-1 by covalent bonding, forming bioreactors. Both will be used to identify active substances in natural and / or synthetic collections using multidimensional chromatographic systems. (AU) | |
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