title: Acetylcholinesterase immobilized capillary reactors coupled to protein coated magnetic beads: a new tool for plant extract ligand screening.
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.
authors.affiliation: Departamento de Química, Universidade Federal de São Carlos, caixa postal 676, São Carlos 13565-905, Brazil; Biomedical Research Center, National Institute on Aging, National Institutes of Health, 251 Bayview Boulevard, Baltimore, MD 21224, USA.
authors.forename: Kenia
authors.initials: K
authors.lastname: Lourenço Vanzolini
authors.forename: Zhengjin
authors.initials: Z
authors.lastname: Jiang
authors.forename: Xiaoqi
authors.initials: X
authors.lastname: Zhang
authors.forename: Lucas Campos Curcino
authors.initials: LC
authors.lastname: Vieira
authors.forename: Arlene Gonçalvez
authors.initials: AG
authors.lastname: Corrêa
authors.forename: Carmen Lucia
authors.initials: CL
authors.lastname: Cardoso
authors.forename: Quezia Bezerra
authors.initials: QB
authors.lastname: Cass
authors.forename: Ruin
authors.initials: R
authors.lastname: Moaddel
mesh_terms: D000110:Acetylcholinesterase / Q000737:chemistry*; D029062:Apocynaceae / Q000737:chemistry*; D019149:Bioreactors; D002800:Cholinesterase Inhibitors / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry; D057166:High-Throughput Screening Assays; D008024:Ligands; D059346:Magnets; D046210:Microfluidic Analytical Techniques; D010936:Plant Extracts / Q000737:chemistry*
chemical_list: D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D008024:Ligands; D010936:Plant Extracts; D000110:Acetylcholinesterase
keywords: Acetylcholinesterase inhibitor; Binding affinities; Non-linear chromatography