Production of bioaffinity columns (afin) for cholinesterase ligands screening

• Grant number: 18/15437-9
• Support type: Research Grants - Innovative Research in Small Business - PIPE
• Duration: March 01, 2020 - November 30, 2020
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
• Principal researcher: Adriana Ferreira Lopes Vilela
• Grantee: Adriana Ferreira Lopes Vilela
• Company: Adriana Ferreira Lopes Vilela
• CNAE: Fabricação de produtos diversos não especificados anteriormente
• City: Ribeirão Preto
• Assoc. researchers: Juliana Maria de Lima
• Associated scholarship(s): 20/02901-9 - Production of bioaffinity columns (Afin®) for cholinesterase ligands screening, BP.PIPE

Abstract

The development of methodologies for screening ligands in the search for compounds with biological activity has been explored to replace the enzymatic assays in solution, which present several limitations such as the low compatibility of the enzymes in organic solvents, high cost and destructible assays. In this context, zonal bioaffinity chromatography (linear and non-linear) has stood out as an extremely powerful tool to be applied in high throughput screening contributing to the identification and characterization of the ligands. Bioaffinity columns are produced by the immobilization of a biological target (enzyme, proteins, antibodies, etc.) in a chromatographic support. Immobilization of enzymes increases its stability in the presence of denaturing factors such as organic solvents and temperature variations, besides allowing the reuse and the use of small amounts of enzyme, corroborating for an excellent relation cost-benefit analysis of enzymatic assays by bioaffinity. The different methods of immobilization and the wide range of available chromatographic supports increase the possibilities of using the IMERs, (immobilized enzyme reactors) in on-line studies thus allowing high productivity of the screening tests with reproducible and reliable results. In this project will be developed bioinfinity columns (Afin®) with the enzymes cholinesterases, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). They are crucial in transmitting nerve impulses in mammals and have received increasing attention in the search for ligands due to their involvement in disorders of the central nervous system such as Alzheimer's disease (AD). The use of commercial cholinesterases for the assays in solution are expensive and with difference in activity between the lots of the enzymes. The production for the commercialization of the Afin® columns proposed here will provide the possibility of non-destructible enzymatic assays making this format a very attractive device for research groups at universities / research centers / and pharmaceutical industries, that working with screening of ligands / inhibitors with anticholinesterase activity. In this context, research in the field of drug and / or insecticide development will be the target audience. The analyzes of the Afin® columns for the quality control will be carried out by zonal elution by liquid chromatography and mass spectrometry. Thus, it will count with the support and collaboration of the Laboratory of the Group of Chromatography of Bioaffinity and Natural Products - GCBPN, Department of Chemistry of the Faculty of Sciences and Letters of Ribeirão Preto of the University of São Paulo, and the laboratory of the Nucleus of Research in Chromatography - SEPARARE at the Federal University of São Carlos. The future company responsible for the production and marketing of Afin® columns can be considered a spin-off. (AU)