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1.
Talanta ; 217: 121093, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32498906

RESUMO

For the first time, we have successfully synthesized stable graphene nanosheets from graphite powder through sonication in the hemoglobin-capped gold nanoclusters (Hb@AuNCs) solution for biosensing application. This approach, as a simple method for the exfoliation and fragmentation of graphite in a nanocluster solution, enabled us to produce stable aqueous graphene dispersions at low cost and without the need for hazardous chemicals or tedious experimental procedures. In this method, Hb@AuNCs were used not only as stabilizing agent of graphene through non-covalent bonding, but also as dispersing agent of few-layer graphene nanosheets. The Hb@AuNCs stabilized graphene (Hb@AuNCs-G) was characterized by high resolution transmission electron microscopy (HRTEM), zeta-sizer and Raman spectroscopy. Then, the graphene nanosheets were applied as a novel versatile electrochemical platform for ultrasensitive biosensing of short DNA species of chronic myelogenous leukemia (CML) based on the "signal off" and "signal on" strategies. For this purpose, a single strand DNA (ssDNA) was immobilized on the Hb@AuNCs-G/AuNPs modified electrode surface and acted as the biorecognition element. Methylene blue (MB), as the signaling probe, was then intercalated into the ssDNA. The intercalated MB was liberated upon interaction with the synthetic complementary DNA (cDNA, target), thereby resulting in the apparent reduction of MB redox signal. This designed "signal off" sensing system enabled the voltammetric determination of the target cDNA over a dynamic linear range (DLR) of 0.1 fM to 10 pM with a limit of detection (LOD) of 0.037 fM. In the "signal on" strategy, the response to the cDNA was detected by monitoring the change in the electron transfer resistance (Rct) using the ferro/ferricyanide system as a redox probe. The charge transfer resistance of the probe was found to increase linearly with increasing concentration of target cDNA in the range of 0.1 fM-10 pM with a limit of detection of 0.030 fM. Finally, the selectivity and feasibility of genosensor was evaluated by the analysis of derived nucleotides from mismatched sequences and the clinical samples of patients with leukemia as real samples, respectively.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Ouro/química , Grafite/química , Hemoglobinas/química , Nanopartículas/química , Proteínas Proto-Oncogênicas c-abl/análise , Proteínas Proto-Oncogênicas c-bcr/análise , Humanos , Tamanho da Partícula , Proteínas Proto-Oncogênicas c-abl/genética , Proteínas Proto-Oncogênicas c-bcr/genética , Propriedades de Superfície
2.
Mater Sci Eng C Mater Biol Appl ; 97: 679-687, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30678956

RESUMO

In this study, a high-performance biosensing nanoplatform based on amidoxime-modified polyacrylonitrile nanofibers decorated with Ag nanoparticles (AgNPs-PAN-oxime NFs) is described. The AgNPs-PAN-oxime NFs were prepared by the combination of electrospinning technique and chemical modification of nitrile group in the PAN. The proposed signal amplifiying nanoplatform was applied in the fabrication of an electrochemical aptasensor for the sensitive detection of CA 125 based on aptamer-cDNA duplex and target induced strand displacement recognition mechanism. The aptasensing interface offers high sensitivity and selectivity for detection of tumor marker due to inherent advantages such as high specific surface area of NFs, good conductivity by doping AgNPs into the polymer NFs and especially the ideal selectivity of anti CA 125 aptamer to its target. The electrochemical aptasensor revealed a wide dynamic linear range (DLR) from 0.01 to 350 U mL-1 with a correlation coefficient of 0.991 and limit of detection (LOD) of 0.0042 U mL-1. Additionally, the designed aptasensor showed acceptable selectivity, reproducibility, repeatability and stability. The satisfactory results for determination of CA 125 in serum samples compared to ELISA method (p-value > 0.05) indicated the potential application of aptasensor in clinical monitoring of tumor biomarker for early diagnosis and management of ovarian cancer.


Assuntos
Resinas Acrílicas/química , Aptâmeros de Nucleotídeos/química , Antígeno Ca-125/sangue , Técnicas Eletroquímicas/métodos , Nanofibras/química , Neoplasias Ovarianas/diagnóstico , Técnicas Biossensoriais/métodos , Eletrodos , Feminino , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Oximas/química , Reprodutibilidade dos Testes , Prata/química
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