Acetylcholinesterase affinity-based screening assay on Lippia gracilis Schauer extracts

Author(s)

Vanzolini, K. L. ^[1] ; Sprenger, R. da F. ^[1] ; Leme, G. M. ^[1] ; Moraes, V. R. de S. ^[2] ; Vilela, A. F. L. ^[3] ; Cardoso, C. L. ^[3] ; Cass, Q. B. ^[1]

Affiliation

^[1] Univ Fed Sao Carlos, SEPARARE Dept Quim, Rodovia Washington Luis, Km 235, BR-13565905 Sao Carlos, SP - Brazil

^[2] Univ Fed Sergipe, Dept Quim, Av Marechal Rondon S-N, BR-49100000 Sao Cristovao, SE - Brazil

^[3] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, Grp Cromatog Bioafinidade & Prod Nat, BR-14040901 Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: Journal of Pharmaceutical and Biomedical Analysis; v. 153, p. 232-237, MAY 10 2018.
• Web of Science Citations: 2

Abstract

The use of affinity-based protein assay produced by covalently linking acetylcholinesterase to magnetic beads, followed by chemical characterization of the selective binders using Liquid Chromatography with tandem High-Resolution Mass Spectrometry (LC-HRMS) is herein described for profiling crude aqueous natural product extracts. The fishing assay was first modulated using galanthamine as a reference ligand and then, the assay condition was adjusted for the aqueous leaves extracts obtained from Lippia gracilis Schauer (genotype 201) that was used as the natural combinatory library. From the experiments, a selective binder has been undisclosed with an accurate mass of 449.1131 m/z and identified as eriodictyol 2'-O-glucoside or eriodictyol 3'-O-glucoside. The selectivity of the binding assay was demonstrated, as much as, that erydictiol 7-O-glucoside was not fished, although it was present in the crude aqueous extract. The binding assay platform exhibited high specificity and did not require any sample pretreatment, making it appropriate for profiling binders at natural libraries. (C) 2018 Elsevier B.V. All rights reserved. (AU)

• FAPESP's process: 13/02054-0 - Enzyme immobilization: new tools for ligand screening
• Grantee: Kenia Lourenço Vanzolini
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/01710-1 - Enzyme ligand: new models of screening
• Grantee: Quezia Bezerra Cass
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/24957-0 - Models of binders rapid screening in extracts of natural products for bioconjugation tests and LC-HRMS
• Grantee: Gabriel Mazzi Leme
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50249-8 - Green chemistry: sustainable synthetic methods employing benign solvents, safer reagents, and bio-renewable feedstock
• Grantee: Arlene Gonçalves Corrêa
• Support Opportunities: Research Grants - Research Centers in Engineering Program