RESUMEN
A sensitive chronocoulometric aptasensor for the detection of thrombin has been developed based on gold nanoparticle amplification. The functional gold nanoparticles, loaded with link DNA (LDNA) and report DNA (RDNA), were immobilized on an electrode by thrombin aptamers performing as a recognition element and capture probe. LDNA was complementary to the thrombin aptamers and RDNA was noncomplementary, but could combine with [Ru(NH3)6]³âº (RuHex) cations. Electrochemical signals obtained by RuHex that bound quantitatively to the negatively charged phosphate backbone of DNA via electrostatic interactions were measured by chronocoulometry. In the presence of thrombin, the combination of thrombin and thrombin aptamers and the release of the functional gold nanoparticles could induce a significant decrease in chronocoulometric signal. The incorporation of gold nanoparticles in the chronocoulometric aptasensor significantly enhanced the sensitivity. The performance of the aptasensor was further increased by the optimization of the surface density of aptamers. Under optimum conditions, the chronocoulometric aptasensor exhibited a wide linear response range of 0.1-18.5 nM with a detection limit of 30 pM. The results demonstrated that this nanoparticle-based amplification strategy offers a simple and effective approach to detect thrombin.
Asunto(s)
Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Oro/química , Nanopartículas/química , Trombina/análisis , Animales , Secuencia de Bases , ADN/química , Técnicas Electroquímicas , Humanos , Límite de Detección , Nanopartículas/ultraestructuraRESUMEN
Two novel probe-label-free electrochemical aptasensors based on methylene blue (MB)-anchored graphene oxide (GO) amplification were developed for thrombin (TB) and ATP detection, taking advantage of the specific binding affinity of the aptamer towards the target and the different affinities of GO for MB, ssDNA, dsDNA, and G-quadruplex.
RESUMEN
The focus of this work was on designing a label-free DNA biosensor based on a super-sandwich assay using the electrochemical impedance spectroscopy technique. For this purpose, we designed a signal-up configuration whose linker probes could hybridize with two regions of the target DNA. In this configuration, the presented target DNA would effectively decrease the electron transfer, which would improve the sensitivity of the sensor. Ultimately, we employed gel electrophoresis to further confirm the formation of the proposed super-sandwich structure.