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

TALANTA

卷:116

Article

Vanzolini, Kenia Lourenco^1,2  Jiang, Zhengjin^3,4  Zhang, Xiaoqi^3,4  Curcino Vieira, Lucas Campos¹  Correa, Arlene Goncalvez¹  Cardoso, Carmen Lucia⁵  Cass, Quezia Bezerra¹  Moaddel, Ruin^2,3,4 

[1] Univ Fed Sao Carlos, Dept Quim, BR-13565905 Sao Carlos, SP, Brazil

[2] NIA, Biomed Res Ctr, NIH, Baltimore, MD 21224 USA

[3] Jinan Univ, Dept Pharm, Guangzhou 510632, Guangdong, Peoples R China

[4] Jinan Univ, Guangdong Prov Key Lab Pharmacodynam Constituents, Guangzhou 510632, Guangdong, Peoples R China

[5] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, BR-14040901 Sao Paulo, SP, Brazil

关键词: Binding affinities;    Acetylcholinesterase inhibitor;    Non-linear chromatography;   

DOI  :  10.1016/j.talanta.2013.07.046

来源: Elsevier

【 摘 要 】

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. Published by Elsevier B.V.

【 授权许可】

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