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Enzyme immobilization: new tools for ligand screening

Grant number: 13/02054-0
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): June 01, 2013
Effective date (End): December 30, 2016
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Organic Chemistry
Principal Investigator:Quezia Bezerra Cass
Grantee:Kenia Lourenço Vanzolini
Host Institution: Centro de Ciências Exatas e de Tecnologia (CCET). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
Associated research grant:13/01710-1 - Enzyme ligand: new models of screening, AP.TEM
Associated scholarship(s):15/18504-0 - Rapid screening and identification of peptides on toxins as ligands of acetylcholinesterase, BE.EP.PD


The use of immobilized capillary enzyme reactors and enzymes coated to magnetic beads for ligand screening has been adopted as a new technique of high throughput screening (HTS) assays. In the proposed project, the combination of two immobilization techniques was used for the first time, which ensures greater selectivity and specificity during the ligand screening, provided by the immobilized enzyme. The bioreactor of magnetic nanoparticles allows the use of complex mixtures without prior fractionation, fishing and identifying the ligands. The subsequent connection to the capillary bioreactor allows the calculation of inhibitory constant (Ki), affinity constant (Kd) and the half maximal inhibitory concentration (IC50). The target selected was the enzyme acetylcholinesterase (AChE), which acts in the central nervous system and rapidly hydrolyzes the active neurotransmitter acetylcholine into the inactive compounds choline and acetic acid. As a reference target for the treatment of Alzheimer's disease and development of novel insecticides, the search for new ligands of AChE is stimulated by structural diversity thereof. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
VANZOLINI, KENIA L.; VIEIRA, LUCAS C. C.; CORREA, ARLENE. G.; MOADDEL, RUIN; CASS, QUEZIA B.. Acetylcholinesterase immobilized on modified magnetic beads as a tool for screening a compound library. Microchimica Acta, v. 182, n. 13-14, p. 2209-2213, . (13/02054-0, 13/01710-1)
VANZOLINI, K. L.; SPRENGER, R. DA F.; LEME, G. M.; MORAES, V. R. DE S.; VILELA, A. F. L.; CARDOSO, C. L.; CASS, Q. B.. Acetylcholinesterase affinity-based screening assay on Lippia gracilis Schauer extracts. Journal of Pharmaceutical and Biomedical Analysis, v. 153, p. 232-237, . (13/02054-0, 13/01710-1, 16/24957-0, 14/50249-8)

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