Anna Porfireva

Polyelectrolyte-mediated assembly of multiwalled carbon nanotubes on living yeast cells.

Here we report the three-dimensional assembly of carbon nanotubes on the polyelectrolyte-coated living Saccharomyces cerevisiae cells using the polyelectrolyte-mediated layer-by-layer approach. Synthetic polyelectrolytes poly(allylamine hydrochloride) and poly(sodium 4-styrenesulfonate) were layer-by-layer deposited on the surfaces of the yeast cells followed by the deposition of water-soluble oxidized multiwalled carbon nanotubes (MWNTs) and an additional outermost polyelectrolyte bilayer. This resulted in the fabrication of polyelectrolyte/nanotubes composite coatings on the cell walls of the yeast cells, which could be clearly seen using the conventional optical microscopy. Transmission and scanning electron microscopy was applied to further investigate the composite coatings. Viability of the encapsulated cells was confirmed using the intercellular esterase activity test. Finally, electrochemical studies using voltammetry and electrochemical impedance measurements were performed, indicating that the composite polyelectrolytes/MWNTs coatings sufficiently affect the electron mediation between the encapsulated yeast cells and the artificial electron acceptor, making it possible to distinguish between living and dead cells. The technique described here may find potential application in the development of microelectronic devices, core-shell and hollow composite microparticles, and electrochemical cell-based biosensors.

Impedimetric Aptasensor for Ochratoxin A Determination Based on Au Nanoparticles Stabilized with Hyper-Branched Polymer

An impedimetric aptasensor for ochratoxin A (OTA) detection has been developed on the base of a gold electrode covered with a new modifier consisting of electropolymerized Neutral Red and a mixture of Au nanoparticles suspended in the dendrimeric polymer Botlorn H30®. Thiolated aptamer specific to OTA was covalently attached to Au nanoparticles via Au-S bonding. The interaction of the aptamer with OTA induced the conformational switch of the aptamer from linear to guanine quadruplex form followed by consolidation of the surface layer and an increase of the charge transfer resistance. The aptasensor makes it possible to detect from 0.1 to 100 nM of OTA (limit of detection: 0.02 nM) in the presence of at least 50 fold excess of ochratoxin B. The applicability of the aptasensor for real sample assay was confirmed by testing spiked beer samples. The recovery of 2 nM OTA was found to be 70% for light beer and 78% for dark beer.

Electrochemical DNA sensors based on electropolymerized materials

The use of electropolymerized materials in the DNA sensors is reviewed with particular emphasis on their functions and specific interactions with DNA and oligonucleotides. Polyaniline, pollypyrrole, polythiophenes and polymeric forms of phenazines play significant role in the immobilization and signal transduction of DNA sensors for the detection of hybridization events, DNA-protein and other specific interactions on the sensor surface. The mechanism of electropolymerization and the influence of oligonucleotides are also considered for various types of polymers. The DNA sensor performance is classified in accordance with the biological targets and composition of the surface layer.

EQCM Biosensors Based on DNA Aptamers and Antibodies for Rapid Detection of Prions

Novel affinity biosensors for detecting cellular prions, PrP(C), based on DNA aptamers and antibodies immobilized onto the carbon nanotubes have been designed and compared in accordance with their binding ability and analytical performance. The biosensors allowed us to detect PrP(C) with the limits of detection of 20 to 50 pM.

Aptabodies – New Type of Artificial Receptors for Detection Proteins

We report on a new type of artificial receptor formed by hybridization of two DNA aptamers for human thrombin (aptabody). This aptasensor based on multiwalled carbon nanotubes allowed us to detect thrombin with detection limit of 0.3 nM, which was 3 times better in comparison with conventional aptamer.

Label-free aptasensor for thrombin determination based on the nanostructured phenazine mediator

New aptasensors based on DNA aptamer and polycarboxylated thiacalix[4]arenes in cone, 1,3-alternate and partial cone configurations bearing Neutral Red (NR) at substituents at the lower rim have been developed and applied for thrombin detection. The assembly of the biorecognition layer was optimized by AFM and EIS study to reach the maximal coverage and regular composition of the surface layer. The interaction of the NR groups with thrombin suppressed the electron hopping between oxidized and reduced mediator groups. This regularly decreased the NR peak current and increased the resistance of the charge transfer. The aptasensor makes it possible to detect from 1 nM to 1 μM of thrombin with the detection limit of 0.05-0.5 nM. No effect of the 1000 excess of bovine serum albumin on the signal was observed. The influence of thiacalix[4]arene configuration on the sensitivity of aptasensor signal toward thrombin is discussed.

Electrochemical DNA sensors based on spatially distributed redox mediators: challenges and promises

Abstract DNA and aptasensors are widely used for fast and reliable detection of disease biomarkers, pharmaceuticals, toxins, metabolites and other species necessary for biomedical diagnostics. In the overview, the concept of spatially distributed redox mediators is considered with particular emphasis to the signal generation and biospecific layer assembling. The application of non-conductive polymers bearing redox labels, supramolecular carriers with attached DNA aptamers and redox active dyes and E-sensor concept are considered as examples of the approach announced.

Electrochemical DNA Sensor Based on Carbon Black—Poly(Neutral Red) Composite for Detection of Oxidative DNA Damage

Voltammetric DNA sensor has been proposed on the platform of glassy carbon electrode covered with carbon black with adsorbed pillar[5]arene molecules. Electropolymerization of Neutral Red performed in the presence of native or oxidatively damaged DNA resulted in formation of hybrid material which activity depended on the DNA conditions. The assembling of the surface layer was confirmed by scanning electron microscopy and electrochemical impedance spectroscopy. The influence of DNA and pillar[5]arene on redox activity of polymeric dye was investigated and a significant increase of the peak currents was found for DNA damaged by reactive oxygen species generated by Cu2+/H2O2 mixture. Pillar[5]arene improves the electron exchange conditions and increases the response and its reproducibility. The applicability of the DNA sensor developed was shown on the example of ascorbic acid as antioxidant. It decreases the current in the concentration range from 1.0 μM to 1.0 mM. The possibility to detect antioxidant activity was qualitatively confirmed by testing tera infusion. The DNA sensor developed can find application in testing of carcinogenic species and searching for new antitumor drugs.