Label-free evaluation of small-molecule-protein interaction using magnetic capture and electrochemical detection

Autor(es)

Uliana, Carolina V. ^[1] ; de Oliveira, Tassia R. ^[1] ; Cominetti, Marcia R. ^[2] ; Faria, Ronaldo C. ^[1]

Afiliação do(s) autor(es)

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

^[2] Univ Fed Sao Carlos, Dept Gerontol, BR-13565905 Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: ANALYTICAL AND BIOANALYTICAL CHEMISTRY; v. 411, n. 10, p. 2111-2119, APR 2019.
• Citações Web of Science: 0

Resumo

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 alpha V beta 3 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 mu mol L-1, with limit of detection of 0.26 mu mol L-1. Then MPs coated with V3 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-alpha V beta 3 integrin association constant, which was calculated as 4.3 x 10(7) M-1. The method proposed proved to be a good label-free alternative to ligand-protein interaction studies. (AU)

• Processo FAPESP: 15/24940-8 - Avaliação da eficácia de mudanças estruturais do [10]-gingerol em combinação com o quimioterápico doxorubicina para o tratamento de câncer de mama: estudos in vitro e in vivo
• Beneficiário: Márcia Regina Cominetti
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/22401-0 - Desenvolvimento de imunossensores descartáveis para detecção de biomarcadores da Doença de Alzheimer
• Beneficiário: Tássia Regina de Oliveira
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 15/19890-1 - Desenvolvimento de métodos para diagnóstico de doenças utilizando partículas magnéticas e arranjo de imunossensores em sistema microfluídico descartável
• Beneficiário: Ronaldo Censi Faria
• Modalidade de apoio: Auxílio à Pesquisa - Regular