An antibody-based platform for melatonin quantification

Autor(es)

Brazaca, Lais C. ^[1] ; Bramorski, Camila B. ^[1] ; Cancino-Bernardi, Juliana ^[1] ; Cruz-Machado, Sanseray da Silveira ^[2] ; Markus, Regina P. ^[2] ; Janegitz, Bruno C. ^[3] ; Zucolotto, Valtencir ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Sao Carlos Inst Phys, Nanomed & Nanotoxicol Grp, BR-13560970 Sao Carlos, SP - Brazil

^[2] Univ Sao Paulo, Inst Biosci, Dept Physiol, Lab Chronopharmacol, BR-05508090 Sao Paulo, SP - Brazil

^[3] Univ Fed Sao Carlos, Dept Nat Sci Math & Educ, BR-13600970 Araras, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: COLLOIDS AND SURFACES B-BIOINTERFACES; v. 171, p. 94-100, NOV 1 2018.
• Citações Web of Science: 1

Resumo

Melatonin, the `chemical signal of darkness', is responsible to regulate biological rhythms and different physiological processes. It is mainly produced by the pineal gland as a hormone in a rhythmic daily basis, but it may also be synthesized by other tissues, such as immune cells, under inflammatory conditions. Its abnormal circulating levels have been related to several diseases such as type 2 diabetes, Alzheimer's disease and some types of cancer. Currently, melatonin is exclusively quantified by ELISA or radioimmunoassays, which although are very sensitive techniques and present low detection limits, usually require specialized personal and equipment, restricting the tests to a limited number of patients. To overcome such limitations, we developed a novel easy-to use electrochemical immunosensor for rapid melatonin quantification. Anti-melatonin antibodies were immobilized into Indium tin oxide (ITO) platforms using (3-Aminopropyl)triethoxysilane (APTES), 1-ethyl-3-(3-dimethylaminopropyl)carboctiimide (EDC) and N-hydroxysuccinimide (NHS) crosslinkers. The platforms were assayed with synthetic and biologically-present melatonin containing samples. The developed device displayed a linear response in the concentration range from 0.75 to 7.5 mu mol/L and a limit of detection of 0.175 mu mol/L using Electrochemical Impedance Spectroscopy (EIS) (R-2 = 0.989) and 0313 mu mol/L using Cyclic Voltammetry (CV) (R-2 = 0.953) for synthetic melatonin. Furthermore, the sensors exhibited a good stability and reproducibility (3,45% and 2.87% for EIS and CV, respectively, n = 3), maintaining adequate response even after 30 days of assembly. On biologically-present melatonin-containing samples the device displayed a similar performance when compared to ELISA technique (deviation of 13.31%). We expect that the developed device contributes significantly to the medical area allowing precise and complete diagnosis of the diseases related to abnormal levels of melatonin. (AU)

• Processo FAPESP: 13/13691-1 - Eixo imune-pineal: integrando a biologia do tempo em condições fisiológicas, fisiopatológicas e patológicas
• Beneficiário: Regina Pekelmann Markus
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 15/02623-0 - Novas Plataformas para o Diagnóstico Rápido e Precoce do Mal de Alzheimer
• Beneficiário: Laís Canniatti Brazaca
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 15/19099-2 - Desenvolvimento de sensores e biossensores eletroquímicos para diversos fins analíticos
• Beneficiário: Bruno Campos Janegitz
• Modalidade de apoio: Auxílio à Pesquisa - Regular