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1.
Biosens Bioelectron ; 80: 674-681, 2016 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-26914375

RESUMEN

The single nucleotide polymorphism (SNP) of the vangl1 gene is highly correlated with Neural Tube Defects (NTDs), a group of severe congenital malformations. It is hindered by the lack of a quantitative detection method. We first propose the use of a DNA biosensor to detect the missense single nucleotide polymorphism (rs4839469 c.346G>A p.Ala116Thr) of the vangl1 gene in this work. Polypyrrole (PPy) and streptavidin were integrated to modify a gold electrode. We took advantage of the PPy's good biocompatibility and excellent conductivity. To further accelerate the electron transfer process at the electrode surface, polyamidoamine dendrimer-encapsulated gold nanoparticles (Au-PAMAM) were used, because Au-PAMAM possess a large number of amino groups to load capture probes (CP). Using the biotin-streptavidin system, the Au-PAMAM-CP bionanocomposite probe, which can detect the target DNA, was conjugated to the electrode surface. Under optimal conditions, the DNA biosensor exhibited a wide linear range of 0.1-100 nM with a low detection limit of 0.033 nM (S/N=3). The results suggest that this approach has the potential to be used in clinical research.


Asunto(s)
Técnicas Biosensibles , Proteínas Portadoras/aislamiento & purificación , ADN/aislamiento & purificación , Proteínas de la Membrana/aislamiento & purificación , Defectos del Tubo Neural/diagnóstico , Proteínas Portadoras/genética , ADN/genética , Oro/química , Humanos , Proteínas de la Membrana/genética , Nanocompuestos/química , Defectos del Tubo Neural/genética , Polímeros/química , Polimorfismo de Nucleótido Simple/genética , Pirroles/química , Estreptavidina/química
2.
Biosens Bioelectron ; 77: 853-9, 2016 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-26516685

RESUMEN

In this work, a novel label-free biosensor was designed for the sensitive and selective determination of Neu5Acα(2-6)Gal ß MP Glycoside using AuPt-PPy(polypyrrole) conductive nanocomposite film as the sensor platform. The introduced AuPt-PPy nanocomposite provided a large surface area for the immobilization of Sambucus nigra agglutinis (SNA) through a coupling agent for specifically recognizing analytes and exhibited high electrocatalytic activity toward the reduction of hydrogen peroxide (H2O2) as an analytical signal. Subsequently, to block the non-specific sites of the modified electrode, GOx was employed instead of the usual sealers. Most importantly, in the presence of glucose, these localized GOx further enhanced the electrochemical signal, which was achieved by the efficient catalysis of glucose. This study is the first that demonstrates the specific detection of Neu5Acα(2-6)Gal ß MP Glycoside using AuPt-PPy as the electrocatalytic. Under optimal conditions, the electrochemical biosensor exhibited a wide linear range of 0.01 pgmL(-1)-800 ngmL(-1) with a low detection limit of 0.003 pgmL(-1) (S/N=3), due to the affinity between SNA and Neu5Acα(2-6)Gal ß MP Glycoside. Therefore, the co-catalysis signal amplification approach has considerable potential in clinical applications and is suitable for the quantification of other biomarkers.


Asunto(s)
Conductometría/instrumentación , Disacáridos/análisis , Nanopartículas del Metal/química , Lectinas de Plantas/química , Polímeros/química , Pirroles/química , Sambucus nigra/metabolismo , Materiales Biocompatibles/química , Catálisis , Disacáridos/química , Diseño de Equipo , Análisis de Falla de Equipo , Oro/química , Platino (Metal)/química , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
3.
Biosens Bioelectron ; 72: 237-46, 2015 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-25985199

RESUMEN

A simplified electrochemical multianalyte immunosensor for the simultaneous detection of high sensitivity C-reactive protein (hsCRP) and soluble CD40 ligand (sCD40L) that uses reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) that directly adsorbs metal ions as labels is reported. rGO-TEPA contains a large number of amino groups and has excellent conductivity, making it an ideal template for the loading of Pb(2+) and Cu(2+), which greatly amplifies the detection signals. The signals could be directly detected in a single run through differential pulse voltammetry (DPV), and each biorecognition event produces a distinct voltammetric peak. The position and size of each peak reflects the identity and the level of the corresponding antigen. Primarily designed for an application in a sandwich-type immunoassay based on Pb(2+) and Cu(2+) labels, two main challenges are accomplished with the herein presented nanosheets: fabrication of the template and the amination process for Pb(2+) and Cu(2+) adsorption. To further improve the analytical performance of the immunosensor, Au@bovine serum albumin (BSA) nanospheres synthesized through a "green" synthesis route were used as a sensor platform, which not only provides a biocompatible microenvironment for the immobilization of antibodies but also amplifies the electrochemical signals. Under optimal conditions, hsCRP and sCD40L could be assayed in the range of 0.05 to 100 ng mL(-1) with detection limits of 16.7 and 13.1 pg mL(-1) (S/N=3), respectively. The assay results on clinical serum samples with the proposed immunosensor were in acceptable agreement with those using the standard single-analyte test of the enzyme-linked immunosorbent assay (ELISA). This novel immunosensing system provides a simple, sensitive and low-cost approach for a multianalyte immunoassay.


Asunto(s)
Proteína C-Reactiva/análisis , Ligando de CD40/sangre , Cobre/aislamiento & purificación , Grafito/química , Inmunoensayo/métodos , Plomo/aislamiento & purificación , Compuestos de Amonio Cuaternario/química , Adsorción , Técnicas Biosensibles/métodos , Ligando de CD40/análisis , Cationes Bivalentes/aislamiento & purificación , Técnicas Electroquímicas/métodos , Oro/química , Humanos , Límite de Detección , Modelos Moleculares , Nanosferas/química , Nanosferas/ultraestructura , Oxidación-Reducción , Óxidos/química , Reproducibilidad de los Resultados
4.
Biosens Bioelectron ; 70: 392-7, 2015 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-25845330

RESUMEN

For the first time, a simple, ultrasensitive and label-free electrochemical monocyte chemotactic protein-1 (MCP-1) immunosensor based on orderly oriented conductive wires has been developed. A conductive wire, which is similar to an electron-conducting tunnel, was designed with Au nanoparticles (AuNPs) joined to Au@Pt core-shell microspheres via a cysteamine (CA) crosslinker. To enhance the sensitivity of the immunosensor, Au nanoparticles were electrodeposited onto the gold electrode, and CA was self-assembled via strong Au-S covalent bonds, providing an appropriate surface and promoting electron transfer. Next, Au@Pt core-shell microspheres with large surface area were grafted onto the modified electrode to immobilize more MCP-1 antibodies. MCP-1 is an initiating factor and biomarker of atherosclerotic diseases. Under optimal experimental conditions, differential pulse voltammetry (DPV) current changes were used to detect MCP-1 with a broad linear range of 0.09-360 pg mL(-1) and a low detection limit of 0.03 pg mL(-1) (S/N=3). The proposed immunosensor exhibited good selectivity, reproducibility and reusability. When applied to spiked serum samples, the data for the developed immunosensor were in agreement with an enzyme linked immunosorbent assay, suggesting that the electrochemical immunosensor would be suitable for practical detection.


Asunto(s)
Quimiocina CCL2/sangre , Conductometría/instrumentación , Inmunoensayo/instrumentación , Nanopartículas del Metal/química , Nanocables/química , Conductividad Eléctrica , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Nanopartículas del Metal/ultraestructura , Microquímica , Nanocables/ultraestructura , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
5.
Biosens Bioelectron ; 69: 95-9, 2015 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-25706062

RESUMEN

In this work, a novel electrochemical immunosensor for the detection of secretoneurin (SN), which uses metal ion functionalised reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) as a label, is reported for the first time. rGO-TEPA contains a large number of amino groups, which makes it an ideal templet for the loading of metal ions. rGO-TEPA-Pb(2+) was employed to immobilise secondary secretoneurin (SN) antibody (Ab2), and the resulting nanocomposite (Ab2-rGO-TEPA-Pb(2+)) was used as a trace tag for signal amplification. A modified electrode consisting of functionalised graphene nanosheets (Au@GS) was used as a substrate to immobilise the antibodies. Under the optimal conditions, the immunoassay exhibited high sensitivity, acceptable stability and reproducibility with a wide linear range from 0.001 to 100ngmL(-1) (R=0.996), and an ultra-low detection limit of 0.33pgmL(-1) (S/N=3). Furthermore, the immunosensor could be employed to detect SN in clinical serum samples. The proposed sensing strategy enriches the electrochemical immunoassay and exhibits potential for the point-of-care diagnostic application of the clinical screening of biomarkers.


Asunto(s)
Conductometría/instrumentación , Etilenodiaminas/química , Grafito/química , Inmunoensayo/instrumentación , Plomo/química , Neuropéptidos/sangre , Secretogranina II/sangre , Electrodos , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Microquímica/instrumentación , Neuropéptidos/química , Oxidación-Reducción , Óxidos/química , Reproducibilidad de los Resultados , Secretogranina II/química , Sensibilidad y Especificidad , Coloración y Etiquetado/métodos
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