Electrochemical methods for DNA hybridization detection have many advantages that are very fast to detect hybridization and can be directly applied for a portable DNA sensor. In this paper, an electrochemical method to directly detect DNA hybridization was developed on the basis of a new conductive polymer, which was polymerized on the glassy carbon electrode with a terthiophene monomer having a carboxyl group (3'-carboxyl-5,2',5',2"-terthiophene). The ss-DNA probe was made by chemically bonding an amine-linked C6 alkyl group to the 5' terminus of oligonucleotide (19-mer). The probe moiety was immobilized on the polymer through covalent bonding with a catalyst, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. A difference in admittance was observed before and after hybridization as a result of the reduction of the resistance after hybridization. The highest difference in admittance was observed around 1 kHz before and after hybridization. Hybridization amounts of end two-base and center one-base mismatched sequences were obtained only in a 14.3% response when compared to that for the complementary matched sequence.
Human epidermal growth factor receptor 2 (HER2) and HER2-overexpressing breast cancer cells were detected using an electrochemical immunosensor combined with hydrazine and aptamer-conjugated gold nanoparticles (AuNPs). The sensor probe was fabricated by covalently immobilizing anti-HER2 onto a nanocomposite layer that was composed of self-assembled 2,5-bis(2-thienyl)-1H-pyrrole-1-(p-benzoic acid) (DPB) on AuNPs. The hydrazine-AuNP-aptamer bioconjugate, where the hydrazine reductant was directly attached onto AuNPs to avoid the nonspecific deposition of silver on the sensor surface, was designed and used to reduce silver ion for signal amplification selectively. The silver-stained target cells were visualized easily by the bare eye and an optical microscope, and the cells were quantitatively analyzed using stripping voltammetry. The parameters affecting the analytical response were optimized. The proposed sensor was capable of differentiating between HER2-positive breast cancer cells and HER2-negative cells. This method exhibited an excellent diagnosis method for the ultrasensitive detection of SK-BR-3 breast cancer cells in human serum samples with a detection limit of 26 cells/mL.
The functionalized conducting polymer (CP) of 5, 2':5', 2' '-terthiophene-3'-carboxylic acid on a platinum microelectrode was prepared through the electropolymerization process using cyclic voltammetry and was used as a substrate for the immobilization of enzymes. The nanoparticles of the CP were obtained at a high scan rate in the cyclic voltammetric experiment. A needle-type amperometric glutamate microbiosensor based on the covalent immobilization of glutamate oxidase (GlOx) onto the CP layer was fabricated for in vivo measurements. The surfaces of the CP/Pt and GlOx/CP/Pt were characterized by QCM, ESCA, and AFM. The biosensor efficiently detected glutamate through the oxidation of enzymatically generated H2O2 at approximately +0.45 V versus Ag/AgCl. Various experimental parameters, such as pH, temperature, and the applied potential in the detection step were optimized. The interference effects from other biological compounds were examined, and ascorbate and dopamine interferences were observed, which were completely minimized by coimmobilizing ascorbate oxidase and by coating the sensor surface with a cationic polymer, polyethyleneimine. A linear calibration plot for glutamate was obtained between 0.2 and 100 microM with a detection limit of 0.1 +/- 0.03 microM. The proposed glutamate microbiosensor was successfully used for in vivo monitoring of the extracellular glutamate released by cocaine stimulation.
The earlier stage of the polymerization reaction of polyaniline has been studied employing in situ spectroelectrochemical techniques. The results indicate that the nitrene cation appears to be a key intermediate species, which leads to all three possible dimers including a head‐to‐tail dimer (N‐phenyl‐p‐phenylenediamine), a tail‐to‐tail dimer (benzidine), and a head‐to‐head dimer (hydrazobenzene). The oxidized forms of these dimers were all shown to be capable of growing polyaniline in the presence of aniline, even though aniline was not oxidized. The cyclic voltammetric peaks observed during the PA synthesis in the middle potential regions were shown to arise from the redox reactions of these dimers, oligomers, and degradation products of polyaniline including quinoneimines and p‐benzoquinone.
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