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Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: A new tool for plant extract ligand screening

Lourenço Vanzolini, Kenia, Jiang, Zhengjin, Zhang, Xiaoqi, Curcino Vieira, Lucas Campos, Gonçalvez Corrêa, Arlene, Lucia Cardoso, Carmen, Bezerra Cass, Quezia, Moaddel, Ruin
Talanta 2013 v.116 pp. 647-652
Alzheimer disease, acetylcholinesterase, bioreactors, central nervous system, chromatography, insecticides, ligands, plant extracts, screening
The use of immobilized capillary enzyme reactors (ICERs) and enzymes coated to magnetic beads ((NT or CT)-MB) for ligand screening has been adopted as a new technique of high throughput screening (HTS). In this work the selected target was the enzyme acetylcholinesterase (AChE), which acts on the central nervous system and is a validated target for the treatment of Alzheimer’s disease, as well as for new insecticides. A new approach for the screening of plant extracts was developed based on the ligand fishing experiments and zonal chromatography. For that, the magnetic beads were used for the ligand fishing experiments and capillary bioreactors for the activity assays. The latter was employed also under non-linear conditions to determine the affinity constants of known ligands, for the first time, as well as for the active fished ligand.