Acetylcholinesterase immobilized on modified magnetic beads as a tool for screening a compound library

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• 作者：Kenia L. Vanzolini ; Lucas C. C. Vieira ; Arlene. G Corrêa ; Ruin Moaddel…
• 关键词：Binding affinity assay ; Acetylcholinesterase inhibitor ; Label ; free assay ; Fishing assay ; Ligand fishing ; Coumarin library
• 刊名：Microchimica Acta
• 出版年：2015
• 出版时间：October 2015
• 年：2015
• 卷：182
• 期：13-14
• 页码：2209-2213
• 全文大小：477 KB
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• 作者单位：Kenia L. Vanzolini (1)
  Lucas C. C. Vieira (1)
  Arlene. G Corrêa (1)
  Ruin Moaddel (2)
  Quezia B. Cass (1)
  
  1. SEPARARE Núcleo de Pesquisa em Cromatografia. Departamento de Química, Universidade Federal de S?o Carlos, Caixa postal 676, S?o Carlos, 13565-905, Brazil
  2. Biomedical Research Center, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd, Baltimore, MD, 21224, USA
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Inorganic Chemistry
  Physical Chemistry
  Characterization and Evaluation Materials
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Wien
• ISSN：1436-5073

文摘

Acetylcholinesterase (AChE) from Electrophorus electricus was immobilized on the surface of amino-modified magnetic beads (AChE-MB), and its activity evaluated by the quantification of acetylcholine hydrolysis. A reference mixture composed of AChE binders (galanthamine and a probe coumarin, K _i--.031?±-.010 μM) and non-binders (ketamine and propranolol) was used to probe the fishing assay. The performance of the bioconjugation assay was demonstrated with a library of 12 reference coumarins from which two ligands were directly identified by LC-MS/MS in a single assay, demonstrating the usefulness of this approach.