Electrochemical DNA-Sensor Based on Macrocyclic Dendrimers with Terminal Amino Groups and Carbon Nanomaterials.

The assembling of thiacalix[4]arene-based dendrimers in cone, partial cone, and 1,3-alternate configuration on the surface of a glassy carbon electrode coated with carbon black or multiwalled carbon nanotubes has been characterized using cyclic voltammetry, electrochemical impedance spectroscopy, and scanning electron microscopy. Native and damaged DNA were electrostatically accumulated on the modifier layer. The influence of the charge of the redox indicator and of the macrocycle/DNA ratio was quantified and the roles of the electrostatic interactions and of the diffusional transfer of the redox indicator to the electrode interface indicator access were established. The developed DNA sensors were tested on discrimination of native, thermally denatured, and chemically damaged DNA and on the determination of doxorubicin as the model intercalator. The limit of detection of doxorubicin established for the biosensor based on multi-walled carbon nanotubes was equal to 1.0 pM with recovery from spiked human serum of 105-120%. After further optimization of the assembling directed towards the stabilization of the signal, the developed DNA sensors can find application in the preliminary screening of antitumor drugs and thermal damage of DNA. They can also be applied for testing potential drug/DNA nanocontainers as future delivery systems.

Impedimetric DNA Sensor Based on Electropolymerized N-Phenylaminophenothiazine and Thiacalix[4]arene Tetraacids for Doxorubicin Determination.

Electrochemical DNA sensors are highly demanded for fast and reliable determination of antitumor drugs and chemotherapy monitoring. In this work, an impedimetric DNA sensor has been developed on the base of a phenylamino derivative of phenothiazine (PhTz). A glassy carbon electrode was covered with electrodeposited product of PhTz oxidation obtained through multiple scans of the potential. The addition of thiacalix[4]arene derivatives bearing four terminal carboxylic groups in the substituents of the lower rim improved the conditions of electropolymerization and affected the performance of the electrochemical sensor depending on the configuration of the macrocyclic core and molar ratio with PhTz molecules in the reaction medium. Following that, the deposition of DNA by physical adsorption was confirmed by atomic force microscopy and electrochemical impedance spectroscopy. The redox properties of the surface layer obtained changed the electron transfer resistance in the presence of doxorubicin due to its intercalating DNA helix and influencing charge distribution on the electrode interface. This made it possible to determine 3 pM-1 nM doxorubicin in 20 min incubation (limit of detection 1.0 pM). The DNA sensor developed was tested on a bovine serum protein solution, Ringer-Locke's solution mimicking plasma electrolytes and commercial medication (doxorubicin-LANS) and showed a satisfactory recovery rate of 90-105%. The sensor could find applications in pharmacy and medical diagnostics for the assessment of drugs able to specifically bind to DNA.

Electrochemical DNA Sensor Based on Acridine Yellow Adsorbed on Glassy Carbon Electrode.

Electrochemical DNA sensors offer unique opportunities for the sensitive detection of specific DNA interactions. In this work, a voltametric DNA sensor is proposed on the base of glassy carbon electrode modified with carbon black, adsorbed acridine yellow and DNA for highly sensitive determination of doxorubicin antitumor drug. The signal recorded by cyclic voltammetry was attributed to irreversible oxidation of the dye. Its value was altered by aggregation of the hydrophobic dye molecules on the carbon black particles. DNA molecules promote disaggregation of the dye and increased the signal. This effect was partially suppressed by doxorubicin compensate for the charge of DNA in the intercalation. Sensitivity of the signal toward DNA and doxorubicin was additionally increased by treatment of the layer with dimethylformamide. In optimal conditions, the linear range of doxorubicin concentrations determined was 0.1 pM-1.0 nM, and the detection limit was 0.07 pM. No influence of sulfonamide medicines and plasma electrolytes on the doxorubicin determination was shown. The DNA sensor was tested on two medications (doxorubicin-TEVA and doxorubicin-LANS) and showed recoveries of 102-105%. The DNA sensor developed can find applications in the determination of drug residues in blood and for the pharmacokinetics studies.

Electrochemical DNA Sensors with Layered Polyaniline-DNA Coating for Detection of Specific DNA Interactions.

A DNA sensor has been proposed on the platform of glassy carbon electrode modified with native DNA implemented between two electropolymerized layers of polyaniline. The surface layer was assembled by consecutive stages of potentiodynamic electrolysis, DNA drop casting, and second electrolysis, which was required for capsulation of the DNA molecules and prevented their leaching into the solution. Surface layer assembling was controlled by cyclic voltammetry, electrochemical impedance spectroscopy, atomic force, and scanning electron microscopy. For doxorubicin measurement, the DNA sensor was first incubated in the Methylene blue solution that amplified signal due to DNA intercalation and competition with the doxorubicin molecules for the DNA binding sites. The charge transfer resistance of the inner layer interface decreased with the doxorubicin concentration in the range from 1.0 pM to 0.1 µM (LOD 0.6 pM). The DNA sensor was tested for the analysis of spiked artificial urine samples and showed satisfactory recovery in concentration range of 0.05⁻10 µM. The DNA sensor developed can find application in testing of antitumor drugs and some other DNA damaging factors.