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

乙酰胆碱酯酶固定化毛细管反应器与蛋白质包被磁珠耦合:植物提取物配体筛选的新工具

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作者:Kenia Lourenço Vanzolini, Zhengjin Jiang, Xiaoqi Zhang, Lucas Campos Curcino Vieira, Arlene Gonçalvez Corrêa, Carmen Lucia Cardoso, Quezia Bezerra Cass, Ruin Moaddel

Abstract

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.

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