Electrochemical aptasensors can detect different cancer biomarkers to provide point-of-care diagnosis that is low cost, rapid, specific and sensitive. In this work, we described the development of an electrochemical single-use aptasensor for detection and analysis of vascular endothelial growth factor (VEGF). Gold nanostructured graphite screen-printed electrodes were firstly modified with a mixed monolayer of a primary thiolated DNA aptamer and a spacer thiol, 6-mercapto-1-hexanol. VEGF protein was then incubated with the aptasensor. An enzyme-amplified detection scheme, based on the coupling of a streptavidin-alkaline phosphatase conjugate and secondary biotinylated aptamer was then applied. The enzyme catalyzed the hydrolysis of the electroinactive 1-naphthylphosphate to 1-naphthol; this product is electroactive and has been detected by means of differential pulse voltammetry (DPV). The aptasensor response was found to be linearly related to the target concentration between 0 and 250 nmol L-1 ; The detection limit was 30 nmol L-1 . The performance of the immunoassay in terms of reproducibility and selectivity has been also studied.
A DNA aptasensor for electrochemical detection of vascular endothelial growth factor / Ravalli, A; Rivas, L.; De La Escosura-Muñiz, Alfredo; Pons, J.; Merkoçi, A.; Marrazza, G.. - In: JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY. - ISSN 1533-4880. - STAMPA. - 15:(2015), pp. 3411-3416. [10.1166/jnn.2015.10037]
A DNA aptasensor for electrochemical detection of vascular endothelial growth factor
RAVALLI, ANDREA;
2015
Abstract
Electrochemical aptasensors can detect different cancer biomarkers to provide point-of-care diagnosis that is low cost, rapid, specific and sensitive. In this work, we described the development of an electrochemical single-use aptasensor for detection and analysis of vascular endothelial growth factor (VEGF). Gold nanostructured graphite screen-printed electrodes were firstly modified with a mixed monolayer of a primary thiolated DNA aptamer and a spacer thiol, 6-mercapto-1-hexanol. VEGF protein was then incubated with the aptasensor. An enzyme-amplified detection scheme, based on the coupling of a streptavidin-alkaline phosphatase conjugate and secondary biotinylated aptamer was then applied. The enzyme catalyzed the hydrolysis of the electroinactive 1-naphthylphosphate to 1-naphthol; this product is electroactive and has been detected by means of differential pulse voltammetry (DPV). The aptasensor response was found to be linearly related to the target concentration between 0 and 250 nmol L-1 ; The detection limit was 30 nmol L-1 . The performance of the immunoassay in terms of reproducibility and selectivity has been also studied.File | Dimensione | Formato | |
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