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Nanomaterials (Basel) ; 11(4)2021 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-33921234


In this work, we developed a sandwich DNA-immunosensor for quantification of the methylated tumour suppressor gene O-6-methylguanine-DNA methyltransferase (MGMT), which is a potential biomarker for brain tumours and breast cancer. The biosensor is based on aminated reduced graphene oxide electrode, which is achieved by ammonium hydroxide chemisorption and anti-5-methylcytosine (anti-5mC) as a methylation bioreceptor. The target single-strand (ss) MGMT oligonucleotide is first recognised by its hybridisation with complementary DNA to form double-stranded (ds) MGMT, which is then captured by anti-5mC on the electrode surface due to the presence of methylation. Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and Scanning electron microscopy (SEM) techniques were used to characterise the electrode surface. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques were used for electrochemical measurements. Under optimised conditions, the proposed biosensor is able to quantify a linear range of concentrations of the MGMT gene from 50 fM to 100 pM with a limit of detection (LOD) of 12 fM. The sandwich design facilitates the simultaneous recognition and quantification of DNA methylation, and the amination significantly improves the sensitivity of the biosensor. This biosensor is label-, bisulfite- and PCR-free and has a simple design for cost-efficient production. It can also be tailor-made to detect other methylated genes, which makes it a promising detection platform for DNA methylation-related disease diagnosis and prognosis.

Food Chem ; 355: 129547, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-33773454


The detection of food adulterants and toxicants can prevent a large variety of adverse health conditions for the global population. Through the process of rapid sensing enabled by deploying novel and robust sensors, the food industry can assist in the detection of adulterants and toxicants at trace levels. Sensor platforms which exploit graphene-based nanomaterials satisfy this requirement due to outstanding electrical, optical and thermal properties. The materials' facile conjugation with linkers and biomolecules along with the option for further enhancement using nanoparticles results in highly sensitive and selective sensing characteristics. This review highlights novel applications of graphene derivatives for detection covering three important approaches; optical, electrical (field-effect) and electrochemical sensing. Suitable graphene-based sensors for portable devices as point-of-need platforms are also presented. The future scope of these sensors is discussed to showcase how these emerging techniques will disrupt the food detection sector for years to come.

Análise de Alimentos/métodos , Contaminação de Alimentos/análise , Grafite/química , Eletricidade , Nanotecnologia
Mikrochim Acta ; 187(6): 338, 2020 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-32430539


The published version of this article, unfortunately, contains errors. Corrections in references were incorrectly carried out. Also, the reduction of graphene oxide was carried out between the potential of -1.5 and 0.5 V, instead of 0.5 and 1.5 V.

Mikrochim Acta ; 187(5): 288, 2020 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-32333119


A label-free biosensor is developed for the determination of plasma-based Aß1-42 biomarker in Alzheimer's disease (AD). The platform is based on highly conductive dual-layer of graphene and electrochemically reduced graphene oxide (rGO). The modification of dual-layer with 1-pyrenebutyric acid N-hydroxysuccinimide ester (Pyr-NHS) is achieved to facilitate immobilization of H31L21 antibody. The effect of these modifications were studied with morphological, spectral and electrochemical techniques. The response of the biosensor was evaluated using differential pulse voltammetry (DPV). The data was acquired at a working potential of ~ 180 mV and a scan rate of 50 mV s-1. A low limit of detection (LOD) of 2.398 pM is achieved over a wide linear range from 11 pM to 55 nM. The biosensor exhibits excellent specificity over Aß1-40 and ApoE ε4 interfering species. Thus, it provides a viable tool for electrochemical determination of Aß1-42. Spiked human and mice plasmas were used for the successful validation of the sensing platform in bio-fluidic samples. The results obtained from mice plasma analysis concurred with the immunohistochemistry (IHC) and magnetic resonance imaging (MRI) data obtained from brain analysis. Graphical abstract Schematic representation of the electrochemical system proposed for Aß1-42 determination: (a) modification of graphene screen-printed electrode (SPE) with monolayer graphene oxide (GO) followed by its electrochemical reduction generating graphene/reduced graphene oxide (rGO) dual-layer (b), modification of dual-layer with linker (c), Aß1-42 antibody (H31L21) (d), bovine serum albumin (BSA) (e) and Aß1-42 peptide (f).

Peptídeos beta-Amiloides/sangue , Técnicas Biossensoriais , Técnicas Eletroquímicas , Grafite/química , Fragmentos de Peptídeos/sangue , Animais , Biomarcadores/sangue , Humanos , Camundongos , Estrutura Molecular , Oxirredução
Sensors (Basel) ; 18(1)2018 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-29361679


Clusterin (CLU) has been associated with the clinical progression of Alzheimer's disease (AD) and described as a potential AD biomarker in blood plasma. Due to the enormous attention given to cerebrospinal fluid (CSF) biomarkers for the past couple of decades, recently found blood-based AD biomarkers like CLU have not yet been reported for biosensors. Herein, we report the electrochemical detection of CLU for the first time using a screen-printed carbon electrode (SPCE) modified with 1-pyrenebutyric acid N-hydroxysuccinimide ester (Pyr-NHS) and decorated with specific anti-CLU antibody fragments. This bifunctional linker molecule contains succinylimide ester to bind protein at one end while its pyrene moiety attaches to the carbon surface by means of π-π stacking. Cyclic voltammetric and square wave voltammetric studies showed the limit of detection down to 1 pg/mL and a linear concentration range of 1-100 pg/mL with good sensitivity. Detection of CLU in spiked human plasma was demonstrated with satisfactory recovery percentages to that of the calibration data. The proposed method facilitates the cost-effective and viable production of label-free point-of-care devices for the clinical diagnosis of AD.

Clusterina/análise , Doença de Alzheimer , Biomarcadores , Técnicas Biossensoriais , Eletrodos , Humanos , Limite de Detecção , Pirenos