Carbon Nanotubes Modified Graphite Electrodes for Monitoring of Biointeraction Between 6‐Thioguanine and DNA

In this present study, single‐walled carbon nanotubes (SWCNT) modified disposable pencil graphite electrodes (SWCNT‐PGEs) were developed for the electrochemical monitoring of anticancer drug, and its interaction with double stranded DNA (dsDNA). Under this aim, SWCNT‐PGEs were applied for the first time in the literature to analyse of 6‐Thioguanine (6‐TG), and also to investigate its interaction with DNA by voltammetric and impedimetric methods. The surface morphologies of PGE and SWCNT‐PGE were explored using scanning electron microscopy (SEM) and electrochemical characterization of unmodified/modified electrodes was performed by cyclic voltammetry (CV). Experimental parameters; such as, the concentration of 6‐TG and its interaction time with dsDNA were optimized by using differential pulse voltammetry (DPV). Additionally, the interaction of 6‐TG with dsDNA was studied in case of different interaction times by electrochemical impedance spectroscopy (EIS) in contrast to voltammetric results. The detection limit of 6‐TG was found to be 0.25 μM by SWCNT‐PGE.     

Graphene Oxide Modified Chemically Activated Graphite Electrodes for Detection of microRNA

In our study, graphene oxide (GO) modified graphite electrodes were used for sensitive and selective impedimetric detection of miRNA. After chemical activation of pencil graphite electrode (PGE) surface using covalent agents (CA), GO modification was performed at the surface of chemically activated PGE. Then, CA‐GO‐PGEs were applied for impedimetric miRNA detection. The microscopic and electrochemical characterization of CA‐GO‐PGEs was performed by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). The optimization of experimental conditions; such as GO concentration, DNA probe concentration and miRNA target concentration was performed by using EIS technique. After the hybridization occurred between miRNA‐34a RNA target and its complementary DNA probe, the hybrid was immobilized onto the surface of CA‐GO‐PGEs. Then, the impedimetric detection of miRNA‐DNA hybridization was performed by EIS. The selectivity of our assay was also tested under the optimum experimental conditions.     

Electrochemical Detection of Activated Protein C Using an Aptasensor Based on PAMAM Dendrimer Modified Pencil Graphite Electrodes

Electrochemical aptasensing of APC was carried out using PAMAM dendrimer modified pencil graphite electrodes (PGEs) for the first time herein. Poly(amidoamine) dendrimer having 16 succinamic acid surface groups (generation 2, G2‐PS) modified PGEs were developed, and then were utilized for APC monitoring using differential pulse voltammetry, electrochemical impedance spectroscopy and cyclic voltammetry. The selectivity of single‐use aptasensor was tested against to other proteins; BSA and THR as well as to the affinity of APC binding to different DNA aptamer, or oligonucleotide. Voltammetric APC detection was also explored in a diluted fetal bovine serum resulting with a detection limit DL as 1.5 µg/mL.     

Direct DNA Hybridization on the Single‐Walled Carbon Nanotubes Modified Sensors Detected by Voltammetry and Electrochemical Impedance Spectroscopy

Carboxylic acid functionalized single‐walled carbon nanotubes modified graphite sensors (SWCNT‐PGEs) were developed for electrochemical monitoring of direct DNA hybridization related to specific sequence of Hepatitis B virus, which substantially enhance the electrochemical transduction resulting from guanine oxidation signal comparison to bare PGEs. The performance characteristics of DNA hybridization on disposable CNT‐PGE were explored measuring the guanine signal in terms of optimum analytical conditions; probe and target concentration, hybridization time, and selectivity. The voltammetric results were also complemented with electrochemical impedance spectroscopy (EIS), that was used to characterize the successful construction of carbon nanotubes modification onto the surface of PGEs.     

Tin oxide nanoparticles-polymer modified single-use sensors for electrochemical monitoring of label-free DNA hybridization

In this study, SnO₂ nanoparticles (SNPs)-poly(vinylferrocenium) (PVF⁺) modified single-use graphite electrodes were developed for electrochemical monitoring of DNA hybridization. The surfaces of polymer modified and polymer-SNP modified pencil graphite electrodes (PGEs) were firstly characterized by using SEM analysis. The electrochemical behaviours of these electrodes were also investigated using the differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS) techniques. The polymer-SNP modified PGEs were then tested for the electrochemical sensing of DNA based on the changes at the guanine oxidation signals. Experimental parameters, such as; different modifications in DNA oligonucleotides, DNA probe concentrations were examined to obtain more sensitive and selective electrochemical signals for nucleic acid hybridization. After optimization studies, DNA hybridization was investigated in the case of complementary of hepatitis B virus (HBV) probe, mismatch (MM), and noncomplementary (NC) sequences.     

Electrochemical sensing of silver tags labelled DNA immobilized onto disposable graphite electrodes

An electrochemical approach for the improved electrochemical sensing of DNA was developed in this study based on the oxidation signals of silver and DNA base, guanine by using disposable pencil graphite electrode (PGE) electrodes. The easy surface modification of disposable electrodes PGEs with nucleic acids was performed by passive adsorption using amino linked DNA oligonucleotide attached onto the surface of silver nanoparticles (Ag-NPs). Firstly, the microscopic characterization of silver nanoparticles was investigated by transmission electron microscopy (TEM) and scanning electron microscopy (SEM), and the electrochemical behaviour of these NPs was studied by using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. Then, the overall performance of this novel electrochemical DNA sensing method based on these nanoparticles is studied and discussed in terms of optimum analytical conditions, such as; the effect of DNA concentration, NPs concentration and different buffer solutions, etc. in order to obtain silver and guanine oxidation signals in higher sensitivity and selectivity. The main features related with this electrochemical assay based on silver nanoparticles are discussed and compared with other assays reported in the literatures.     

Shape-controlled electrodeposition of silver using chitosan as structure-directing agent on disposable pencil graphite electrodes: low-cost electrocatalysts for the detection of hydrogen peroxide and hydrazine hydrate

A facile synthetic protocol for the electrodeposition of diverse morphologies of silver on disposable pencil graphite electrodes (Ag/PGE) in the presence of chitosan as structure-directing agent (SDA) is reported. The influence of various electrodeposition parameters on the morphology of Ag deposited has been studied and interpreted using electron microscopic techniques. Several impressive morphologies such as hexahedron, leaf and dendrites have been observed for Ag/PGE with respect to change in experimental conditions. Furthermore, the crucial role of chitosan in determining the morphology of Ag/PGE has been elucidated with the help of three-dimensional Scharifker-Hills nucleation and growth model. The electrocatalytic activities of various Ag/PGEs towards the reduction of hydrogen peroxide (HP) and oxidation of hydrazine hydrate (HH) have been studied in detail with the help of diverse electrochemical techniques. In comparison with PGE, the Ag hexahedron- (Ag-Hex/PGE) and Ag dendrite- (Ag-Dend/PGE) modified PGEs exhibited excellent electrocatalytic activity towards HP and HH, respectively. The Ag-Hex/PGE displayed a wide linear range of 0.1–20,000 μM with a limit of detection (LOD, 3σ/m) of 0.06 μM for HP reduction. On the other hand, a linear range of 25–20,000 μM with LOD of 1.8 μM for HH oxidation has been observed for Ag-Dend/PGE. Furthermore, the modified Ag/PGEs revealed remarkable reproducibility and long-term storage stability. The practical applicability of the Ag-Hex/PGE and Ag-Dend/PGE was demonstrated through the electrocatalytic detection of HP in milk and HH in tap water samples with satisfactory recovery results.

     

Recent Advances in Nanomaterial‐Modified Pencil Graphite Electrodes for Electroanalysis

Pencil graphite electrodes (PGEs) have several advantages over other carbon‐based or commercial metal electrodes, including widespread availability, very low cost, and ease of modification. To make the best use of PGEs in electroanalysis, significant recent advances in the development of different nanomaterial‐PGEs have been observed. The literature published up to mid‐2015 is summarized in the present review, with a focus on the various methodologies used to readily modify graphite pencil electrodes using nanomaterials. This review also touches on the surface characterization of these electrodes and their potential applications in a variety of electrochemical detection applications. The review outlines the scope for further research in this area and discusses the importance of surface modifications of conventional PGE electrodes using nanomaterials or a combination of nanomaterials and electroactive polymers.     

Hydroxyapatite Nanoparticles Modified Graphite Electrodes for Electrochemical DNA Detection

Hydroxyapatite nanoparticles (HaNP) modified pencil graphite electrodes (PGEs) were developed for the first time in the literature, and accordingly they were applied for electrochemical monitoring of sequence‐selective DNA hybridization. The experimental conditions for HaNP modification of PGE, and DNA hybridization related to Hepatitis B Virus (HBV) DNA sequence were optimized. The microscopic and electrochemical characterization of HaNP‐PGE in contrast to the unmodified one was utilized. Under optimized experimental conditions, the selectivity of HBV DNA probe immobilized biosensor was tested against to non‐complementary (NC), mismatch (MM) sequences and the mixture of target:NC (1 : 1) or target: MM (1 : 1).     

Multiwalled Carbon Nanotubes‐Chitosan Modified Single‐Use Biosensors for Electrochemical Monitoring of Drug‐DNA Interactions

A multiwalled carbon nanotubes (CNT)‐chitosan (CHIT) modified pencil graphite electrode (CNT‐CHIT/PGE) was developed for the first time herein for electrochemical monitoring of the interaction of an anticancer drug, mitomycin C (MC) and DNA. The characterization of unmodified PGE, CHIT/PGE, CNT/PGE and CHIT‐CNT/PGE were performed by scanning electron microscopy and cyclic voltammetry techniques. The oxidation signals of MC and guanine were measured before and after interaction at the surface of CNT‐CHIT/PGEs using differential pulse voltammetry. Electrochemical impedance spectroscopy technique was also successfully utilized for monitoring of the interaction process at the surface of CNT‐CHIT/PGEs in different interaction times.     

Carbon nanotube-chitosan modified disposable pencil graphite electrode for vitamin B12 analysis

A single walled carbon nanotube-chitosan (SWCNT-chitosan) modified disposable pencil graphite electrode (PGE) was used in this study for the electrochemical detection of Vitamin B(12). Electrochemical behaviors of SWCNT-chitosan PGE and chitosan modified PGE were compared by using cyclic voltammetry (CV), square-wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS) techniques. SWCNT-chitosan modified electrode was also used for the quantification of Vitamin B(12) in pharmaceutical products. The results show that this electrode system is suitable for sensitive Vitamin B(12) analysis giving good recovery results. The surface morphologies of the SWCNT-chitosan PGE, chitosan modified PGE and unmodified PGE were characterized by using scanning electron microscopy (SEM).     

Interaction of Mitomycin C with DNA Immobilized onto Single‐walled Carbon Nanotube/Polymer Modified Pencil Graphite Electrode

The electrochemical investigation of the interaction between the anticancer drug mitomycin C (MC) and DNA was described using a single‐walled carbon nanotube (SWCNT)/poly(vinylferrocenium) (PVF⁺) modified pencil graphite electrode (PGE). The electrochemical oxidation signals of guanine were monitored before and after the interaction between MC and DNA by using differential pulse voltammetry. The effects of DNA and MC concentration and MC interaction time were examined based on the electrode response. Cyclic voltammetry and electrochemical impedance spectroscopy were used for the characterization of SWCNT/PVF⁺ modified and PVF⁺ modified PGEs. The detection limit corresponded to 625 ng/mL for MC using calf thymus double‐stranded DNA immobilized SWCNT/PVF⁺ modified PGE.     

Ionic Liquid Modified Single‐use Electrode Developed for Voltammetric Detection of miRNA‐34a and its Application to Real Samples

The ionic liquid (IL) modified chemically activated (CA) pencil graphite electrodes (PGEs) were developed for label‐free voltammetric detection of miRNA‐34a, and implemented to the real samples. Firstly, the electrochemical characterization of unmodified PGE, CA‐PGE, IL‐PGE and IL‐CA‐PGE was performed by cyclic voltammetry (CV) as well as their DNA binding capacity was studied by electrochemical impedance spectroscopy (EIS) technique. The microscopic characterization of the surface of each electrodes was investigated by scanning electron microscopy (SEM). Differential pulse voltammetry (DPV) technique was used for measuring the oxidation signal of guanine in order to perform a label‐free voltammetric monitoring of a full‐match hybridization specific to miRNA‐34a. The selectivity of biosensor was tested against to miRNA‐155, miRNA‐660 as well as to the mismatch sequence of miRNA‐34a. The further selectivity of this proposed biosensor was studied in the mixture of samples containing miRNA‐34a with other miRNAs (1 : 1). The voltammetric detection of miRNA‐34a was also explored in the artificial serum medium as fetal bovine serum (FBS) and also in total RNA samples isolated from HUH‐7 human hepatocellular carcinoma cell line.     

Electrochemical probe DNA design in PCR amplicon sequence for the optimum detection of microbiological diseases

Direct electrochemical genosensor was developed for the detection of a probe sequence relative position in a PCR amplicon for the optimum detection of bacterial and microbiological diseases, in this study. The genosensor relies on a label-free electrochemical detection. The amino-linked inosine modified (guanine-free) coequal capture probes which were chosen from different parts of a PCR amplicon, immobilized on to disposable pencil graphite electrodes (PGE) by electrostatically and covalently. As a model case Hepatitis B virus (HBV) genome amplicon was used for the detection and specification. Hybridization was occurred after surface coverage with denatured amplicons. After hybridization, optimum probe sequence position was identified by using the differences between the responses of guanine oxidation signals. The results of this study might have a great convenience for the microbiological diseases detection applications such as DNA micro arrays.     

Indicator-based and indicator-free magnetic assays connected with disposable electrochemical nucleic acid sensor system

An indicator-based and indicator-free magnetic assays connected with a disposable pencil graphite electrode (PGE) were successfully developed, and also compared for the electrochemical detection of DNA hybridization. The oxidation signals of echinomycin (ECHI) and electroactive DNA bases, guanine and adenine, respectively were monitored in the presence of DNA hybridization by using differential pulse voltammetry (DPV) technique. The biotinylated probe was immobilized onto the magnetic beads (magnetic particles, microspheres) and hybridization with its complementary target at the surface of particles within the medium was exhibited successfully using electrochemical sensor system. For the selectivity studies, the results represent that both indicator-based and indicator-free magnetic assays provide a better discrimination for DNA hybridization compared to duplex with one-base or more mismatches. The detection limits (S/N = 3) of the magnetic assays based on indicator or indicator-free were found in nM concentration level of target using disposable sensor technology with good reproducibility. The characterization and advantages of both proposed magnetic assays connected with a disposable electrochemical sensor are also discussed and compared with those methods previously reported in the literature.     

Detection of p53 Gene by Using Genomagnetic Assay Combined with Carbon Nanotube Modified Disposable Sensor Technology

Electrochemical monitoring of DNA hybridization related to p53 gene sequence was investigated using genomagnetic assay combined with single walled carbon nanotube (SWCNT) modified pencil graphite electrodes (PGEs). The hybridization was performed either at magnetic beads (MB) surface or in solution. The enhanced guanine signal was obtained using SWCNT‐PGEs compared to one obtained by unmodified PGEs. The selectivity of genomagnetic assay was tested under optimum conditions. The DLs were calculated as 0.88 µM and 0.11 µM for hybridization performed at MB surface and solution, respectively. This selective, practical and cost effective genomagnetic assay combined with SWCNT‐PGEs is reported herein for the first time.     

Open‐circuit Electrochemical Polymerization for the Sensitive Detection of Phenols

We report on a novel electrochemical method for electro‐polymerizing phenols under open‐circuit conditions. The method developed here is simple, sensitive, rapid, and overcomes the well‐documented surface fouling of carbon electrodes by phenols. A pre‐charged disposable graphite pencil electrode (pCGPE) was found to be useful for both phenol sampling and sensing. The pCGPE was prepared by charging the surface of a graphite pencil electrode by applying a cyclic voltammetry electrochemical treatment in a NaOH solution. Phenol sampling was accomplished by immersing the pCGPE into a phenol solution. This method permitted phenol detection with a detection limit of 4.17 nM (0.39 ppt).     

Streptavidin Modified Carbon Nanotube Based Graphite Electrode for Label‐Free Sequence Specific DNA Detection

Disposable graphite pencil electrodes (PGE) modified with multiwalled carbon nanotubes (MWCNTs)‐streptavidin (STR) conjugates were used for electrochemical monitoring of label‐free DNA hybridization. The surface morphology of PGE electrode before and after hybridization was characterized by scanning electron microscopy. Electrochemical impedance spectroscopy was used to monitor each step of the construction of the DNA biosensor. The biosensor was demonstrated to have excellent selectivity, being able to differentiate complementary sequences from a noncomplementary ones and in addition select the target sequence of DNA from a mixture of other DNA without loss in current sensitivity.     

Poly(dimethylsiloxane) cross-linked carbon paste electrodes for microfluidic electrochemical sensing

Recently, the development of electrochemical biosensors as part of microfluidic devices has garnered a great deal of attention because of the small instrument size and portability afforded by the integration of electrochemistry in microfluidic systems. Electrode fabrication, however, has proven to be a major obstacle in the field. Here, an alternative method to create integrated, low cost, robust, patternable carbon paste electrodes (CPEs) for microfluidic devices is presented. The new CPEs are composed of graphite powder and a binder consisting of a mixture of poly(dimethylsiloxane) (PDMS) and mineral oil. The electrodes are made by filling channels molded in previously cross-linked PDMS using a method analogous to screen printing. The optimal binder composition was investigated to obtain electrodes that were physically robust and performed well electrochemically. After studying the basic electrochemistry, the PDMS-oil CPEs were modified with multi-walled carbon nanotubes (MWCNT) and cobalt phthalocyanine (CoPC) for the detection of catecholamines and thiols, respectively, to demonstrate the ease of electrode chemical modification. Significant improvement of analyte signal detection was observed from both types of modified CPEs. A nearly 2-fold improvement in the electrochemical signal for 100 μM dithiothreitol (DTT) was observed when using a CoPC modified electrode (4.0 ± 0.2 nA (n = 3) versus 2.5 ± 0.2 nA (n = 3)). The improvement in signal was even more pronounced when looking at catecholamines, namely dopamine, using MWCNT modified CPEs. In this case, an order of magnitude improvement in limit of detection was observed for dopamine when using the MWCNT modified CPEs (50 nM versus 500 nM). CoPC modified CPEs were successfully used to detect thiols in red blood cell lysate while MWCNT modified CPEs were used to monitor temporal changes in catecholamine release from PC12 cells following stimulation with potassium.     

Electrochemical Monitoring of DNA Hybridization by Multiwalled Carbon Nanotube Based Screen Printed Electrodes

The application of multiwalled carbon nanotube (MWCNT) based screen printed graphite electrodes (SPEs) was explored in this study for the electrochemical monitoring of DNA hybridization related to specific sequences on Hepatitis B virus (HBV) DNA. After the microscopic characterization of bare MWCNT‐SPEs and DNA immobilized ones was performed, the optimization of assay has been studied. The development of screen printing process combined with nanomaterial based disposable sensor technology leads herein a great opportunity for DNA detection using differential pulse voltammetry (DPV) by measuring the guanine oxidation signal observed at +1.00 V in the presence of DNA hybridization between HBV probe and its complementary, target. The detection limit estimated for signal to noise ratios =3 corresponds to 96.33 nM target concentration in the 40 μL samples. The advantages of carbon nanotube based screen printed electrode used for electrochemical monitoring of DNA hybridization are discussed with sensitivity, selectivity and reproducibility in comparison with previous nanomaterial based electrochemical transducers developed for DNA or other biomolecular recognitions.