Label-free evaluation of small-molecule-protein interaction using magnetic capture and electrochemical detection.
Anal Bioanal Chem
; 411(10): 2111-2119, 2019 Apr.
Article
em En
| MEDLINE
| ID: mdl-30739194
The evaluation of interaction between small molecules and protein is an important step in the discovery of new drugs and to study complex biological systems. In this work, an alternative method was presented to evaluate small-molecule-protein interaction by using ligand capture by protein-coated magnetic particles (MPs) and disposable electrochemical cells. The interaction study was conducted using [10]-gingerol from ginger rhizome and a transmembrane protein αVß3 integrin. Initially, the electrochemical behavior of the natural compound [10]-gingerol was evaluated with the disposable carbon-based electrodes and presented an irreversible oxidation process controlled by diffusion. The analytical curve for [10]-gingerol was obtained in the range of 1.0 to 20.0 µmol L-1, with limit of detection of 0.26 µmol L-1. Then MPs coated with αVß3 integrin were incubated with standard solutions and extracts of ginger rhizome for [10]-gingerol capture and separation. The bioconjugate obtained was dropped to the disposable electrochemical cells, keeping a permanent magnet behind the working electrode, and the binding process was evaluated by the electrochemical detection of [10]-gingerol. The assay method proposed was also employed to calculate the [10]-gingerol-αVß3 integrin association constant, which was calculated as 4.3 × 107 M-1. The method proposed proved to be a good label-free alternative to ligand-protein interaction studies. Graphical abstract á
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Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Catecóis
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Integrina alfaVbeta3
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Proteínas Imobilizadas
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Descoberta de Drogas
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Técnicas Eletroquímicas
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Álcoois Graxos
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Imãs
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Revista:
Anal Bioanal Chem
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
Brasil
País de publicação:
Alemanha