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1.
Mikrochim Acta ; 187(8): 479, 2020 08 02.
Article in English | MEDLINE | ID: mdl-32740774

ABSTRACT

A novel electrochemical nanobiosensor for the detection of miR-155 (as breast cancer biomarker) is introduced . Fe3O4NPs@Ag core-shell nanoparticles were synthesized and their shape and characteristics were confirmed by scanning electron microscope (SEM) imaging, Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) methods. Synthesized nanoparticles were applied onto the magnetic bar carbon paste electrode and then the amine-modified anti-miR-155 (single-stranded probes) was applied on the modified electrode surface and upon hybridization with target miR-155, resveratrol (RSV) was eventually applied as an electrochemical label on the double-strand oligonucleotide. Differential pulse voltammetry (DPV) of the oxidation peak of RSV was assumed as the final signal by sweeping potential from 0 to 0.6 V (vs. Ag/AgCl). The fabrication process was optimized through a series of experiments and the optimized process was confirmed using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The linear range of the fabricated nanobiosensor was 0.5 fM to 1.0 nM and the detection limit was 0.15 fM. The nanobiosensor was able to pass reproducibility and specificity tests using different types of mismatched target sequences.Spiked real samples of human serum were used to confirm that the nanobiosensor enables detection of miR-155 without any significant interferences from other moieties and molecules. Finally, the molecular dynamics simulation of the RSV interaction with single- and double-stranded oligonucleotide was performed and confirmed the preferential binding of RSV to double-stranded DNA; therefore, it can be used as the electrochemical label of DNA and/or miRNA hybridization-based biosensors. Graphical abstract.


Subject(s)
Biosensing Techniques/methods , DNA, Single-Stranded/chemistry , Magnetite Nanoparticles/chemistry , MicroRNAs/blood , Oligodeoxyribonucleotides/chemistry , Resveratrol/chemistry , Biomarkers, Tumor/blood , Biomarkers, Tumor/genetics , DNA, Single-Stranded/genetics , DNA, Single-Stranded/metabolism , Electrochemical Techniques/methods , Humans , Immobilized Nucleic Acids/chemistry , Immobilized Nucleic Acids/genetics , Immobilized Nucleic Acids/metabolism , Limit of Detection , MicroRNAs/genetics , Molecular Docking Simulation , Nanocomposites/chemistry , Nucleic Acid Hybridization , Oligodeoxyribonucleotides/genetics , Oligodeoxyribonucleotides/metabolism , Reproducibility of Results , Resveratrol/metabolism , Silver/chemistry
2.
Mikrochim Acta ; 185(9): 405, 2018 08 09.
Article in English | MEDLINE | ID: mdl-30094655

ABSTRACT

This paper reports on a sensitive and selective method for the detection of Michigan Cancer Foundation-7 (MCF-7) human breast cancer cells and MUC1 biomarker by using an aptamer-based sandwich assay. A biocompatible nanocomposite consisting of multiwall carbon nanotubes (MWCNT) and poly(glutamic acid) is placed on a glassy carbon electrode (GCE). The sandwich assay relies on the use of a mucin 1 (MUC1)-binding aptamer that is first immobilized on the surface of modified GCE. Another aptamer (labeled with silver nanoparticles) is applied for secondary recognition of MCF-7 cells in order to increase selectivity and produce an amplified signal. Differential pulse anodic stripping voltammetry was used to follow the electrochemical signal of the AgNPs. Under the optimal condition, the sensor responds to MCF-7 cells in the concentration range from 1.0 × 102 to 1.0 × 107 cells·mL-1 with a detection limit of 25 cells. We also demonstrate that the MUC1 tumor marker can be detected by the present biosensor. The assay is highly selective and sensitive, acceptably stable and reproducible. This warrants the applicability of the method to early diagnosis of breast cancer. Graphical abstract Schematic of the fabrication of an aptamer-based sandwich biosensor for Michigan Cancer Foundation-7 cells (MCF-7). A MWCNT-poly(glutamic acid) nanocomposite was used as a biocompatible matrix for MUC1-aptamer immobilization. Stripping voltammetry analysis of AgNPs was performed using aptamer conjugated AgNPs as signalling probe.


Subject(s)
Aptamers, Nucleotide/metabolism , Biosensing Techniques/methods , Breast Neoplasms/pathology , Nanocomposites/chemistry , Nanotubes, Carbon/chemistry , Polyglutamic Acid/chemistry , Silver/chemistry , Aptamers, Nucleotide/chemistry , Carbon/chemistry , Electrochemistry , Electrodes , Humans , Limit of Detection , MCF-7 Cells , Metal Nanoparticles/chemistry , Mucin-1/blood , Mucin-1/metabolism , Surface Properties
3.
Mater Sci Eng C Mater Biol Appl ; 75: 1435-1447, 2017 Jun 01.
Article in English | MEDLINE | ID: mdl-28415435

ABSTRACT

In this paper, a highly sensitive voltammetric sensor based on a carbon paste electrode with CuFe2O4 nanoparticle (RGO/CuFe2O4/CPE) was designed for determination of hydrogen peroxide (H2O2). The electrocatalytic reduction of H2O2 was examined using various techniques such as cyclic voltammetry (CV), chronoamperometry, amperometry and differential pulse voltammetry (DPV). CuFe2O4 nanoparticles were synthesized by co-precipitation method and characterized with scanning electron microscopy (SEM), Transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) techniques. Then, a high conductive platform based on a carbon paste electrode modified with RGO and CuFe2O4 nanoparticles was prepared as a suitable platform for determination of hydrogen peroxide. Under the optimum conditions (pH5), the modified electrode indicated a fast amperometric response of <2s, good linear range of 2 to 200µM, low detection limit of 0.52µM for determination of hydrogen peroxide. Also, the peak current of differential pulse voltammetry (DPV) of hydrogen peroxide is increased linearly with its concentration in the ranges of 2 to 10µM and 10 to 1000µM. The obtained detection limit for hydrogen peroxide was evaluated to be 0.064µM by DPV. The designed sensor was successfully applied for the assay of hydrogen peroxide in biological and pharmaceutical samples such as milk, green tea, and hair dye cream and mouthwash solution.


Subject(s)
Copper/chemistry , Electrochemical Techniques/methods , Ferrous Compounds/chemistry , Graphite/chemistry , Hydrogen Peroxide/analysis , Electrodes
4.
Mikrochim Acta ; 185(1): 50, 2017 12 14.
Article in English | MEDLINE | ID: mdl-29594398

ABSTRACT

This article describes an impedimetric aptasensor for the prostate specific antigen (PSA), a widely accepted prostate cancer biomarker. A glassy carbon electrode (GCE) was modified with titanium oxide nanoparticles (TiO2) and silk fibroin nanofiber (SF) composite. The aptasensor was obtained by immobilizing a PSA-binding aptamer on the AuNP-modified with 6-mercapto-1-hexanol. The single fabrication steps were characterized by cyclic voltammetry and electrochemical impedance spectroscopy. The assay has two linear response ranges (from 2.5 fg.mL-1 to 25 pg.mL-1, and from 25 pg.mL-1 to 25 ng.mL-1) and a 0.8 fg.mL -1 detection limit. After optimization of experimental conditions, the sensor is highly selective for PSA over bovine serum albumin and lysozyme. It was successfully applied to the detection of PSA in spiked serum samples. Graphical abstract Schematic of the fabrication of an aptasensor for the prostate specific antigen (PSA). It is based on the use of a glassy carbon electrode modified with gold nanoparticles and titanium oxide-silk fibroin. The immobilization process of aptamer and interaction with PSA were followed by electrochemical impedance spectroscopy technique.


Subject(s)
Aptamers, Nucleotide/chemistry , Electrochemical Techniques/methods , Prostate-Specific Antigen/analysis , Prostatic Neoplasms/diagnosis , Electrochemical Techniques/standards , Electrodes , Fibroins , Humans , Male , Metal Nanoparticles/chemistry , Molecular Probes/chemistry , Molecular Probes/standards , Nanofibers/chemistry , Spectrum Analysis , Titanium
5.
Talanta ; 147: 621-7, 2016 Jan 15.
Article in English | MEDLINE | ID: mdl-26592654

ABSTRACT

In this research, we have developed lable free DNA biosensors based on modified glassy carbon electrodes (GCE) with reduced graphene oxide (RGO) and carbon nanotubes (MWCNTs) for detection of DNA sequences. This paper compares the detection of BRCA1 5382insC mutation using independent glassy carbon electrodes (GCE) modified with RGO and MWCNTs. A probe (BRCA1 5382insC mutation detection (ssDNA)) was then immobilized on the modified electrodes for a specific time. The immobilization of the probe and its hybridization with the target DNA (Complementary DNA) were performed under optimum conditions using different electrochemical techniques such as cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The proposed biosensors were used for determination of complementary DNA sequences. The non-modified DNA biosensor (1-pyrenebutyric acid-N- hydroxysuccinimide ester (PANHS)/GCE), revealed a linear relationship between ∆Rct and logarithm of the complementary target DNA concentration ranging from 1.0×10(-16)molL(-1) to 1.0×10(-10)mol L(-1) with a correlation coefficient of 0.992, for DNA biosensors modified with multi-wall carbon nanotubes (MWCNTs) and reduced graphene oxide (RGO) wider linear range and lower detection limit were obtained. For ssDNA/PANHS/MWCNTs/GCE a linear range 1.0×10(-17)mol L(-1)-1.0×10(-10)mol L(-1) with a correlation coefficient of 0.993 and for ssDNA/PANHS/RGO/GCE a linear range from 1.0×10(-18)mol L(-1) to 1.0×10(-10)mol L(-1) with a correlation coefficient of 0.985 were obtained. In addition, the mentioned biosensors were satisfactorily applied for discriminating of complementary sequences from noncomplementary sequences, so the mentioned biosensors can be used for the detection of BRCA1-associated breast cancer.


Subject(s)
BRCA1 Protein/genetics , Biosensing Techniques , Carbon/chemistry , DNA, Single-Stranded/chemistry , Electric Impedance , Electrochemical Techniques , Electrodes , Graphite/chemistry , Limit of Detection , Mutation , Nanotubes, Carbon/chemistry , Nucleic Acid Hybridization , Oxides/chemistry
6.
Anal Biochem ; 484: 24-30, 2015 Sep 01.
Article in English | MEDLINE | ID: mdl-25988596

ABSTRACT

We have designed a simple and novel electrochemical biosensor based on glassy carbon electrode (GCE) for DNA detection. GCE was modified with reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) by the electrochemical method, which is helpful for immobilization of thiolated bioreceptors. The electrode modification processes were characterized by scanning electron microscopy (SEM) and electrochemical methods. Then a single-stranded DNA (ssDNA) probe for BRCA1 5382 insC mutation detection was immobilized on the modified electrode for a specific time. The experimental conditions, such as probe immobilization time and target DNA (complementary DNA) hybridization time and temperature with probe DNA, were optimized using electrochemical methods. The electrochemical response for DNA hybridization and synthesis was measured using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) methods. The calibration graph contains two linear ranges; the first part is in the range of 3.0×10(-20) to 1.0×10(-12)M, and the second segment part is in the range of 1.0×10(-12) to 1.0×10(-7)M. The biosensor showed excellent selectivity for the detection of the complementary sequences from noncomplementary sequences, so it can be used for detection of breast cancer.


Subject(s)
Biosensing Techniques/methods , DNA/analysis , Glass/chemistry , Gold/chemistry , Graphite/chemistry , Limit of Detection , Oxides/chemistry , BRCA1 Protein/genetics , Base Sequence , Biosensing Techniques/instrumentation , DNA/biosynthesis , DNA/chemistry , DNA/genetics , DNA Mutational Analysis , DNA Probes/chemistry , DNA Probes/genetics , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/genetics , DNA-Directed DNA Polymerase/metabolism , Electrochemistry , Electrodes , Nucleic Acid Hybridization , Oxidation-Reduction , Reproducibility of Results
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