AIKATERINI MANTZILA

Abstract: The development of a faradic impedimetric immunosensor for the detection of S. typhimurium in milk is described for first time. Polyclonal anti-Salmonella was cross-linked, in the presence of glutaraldehyde, on gold electrodes modified with a single 11-amino-1-undecanethiol (MUAM) self-assembled monolayer (SAM) or a mixed SAM of MUAM and 6-mercapto-1-hexanol at a constant 1 + 3 proportion, respectively. The mixed SAM was also deposited in the presence of triethylamine, which was used to prevent the formation of interplane hydrogen bonds among amine-terminated thiols. The effect of the different surface modifications on both the sensitivity and the selectivity of the immunosensors was investigated. The alteration of the interfacial features of the electrodes due to different modification or recognition steps, was measured by faradic electrochemical impedance spectroscopy in the presence of a hexacyanoferrate(II)/(III) redox couple. A substantial amplification of the measuring signal was achieved by performing the immunoreaction directly in culture samples. This resulted in immunosensors with great analytical features, as follows: (i) high sensitivity; the response of the immunosensors increases with respect to the detection time as a consequence of the simultaneous proliferation of the viable bacteria cells in the tested samples; (ii) validity; the response of the immunosensors is practically insensitive to the presence of dead cells; (iii) working simplicity; elimination of various centrifugation and washing steps, which are used for the isolation of bacteria cells from the culture. The proposed immunosensors were successfully used for the detection of S. typhimurium in experimentally inoculated milk samples. The effect of different postblocking agents on the performance of the immunosensors in real samples was also examined.

Abstract: The development of an immunosensor for the direct probing of the interaction between a cysteine-modified synthetic peptide, which corresponds to the epitope cTnC-89-98 of troponin C, and its specific antibody is described. Following immobilization of the peptide onto gold electrodes through the formation of a self-assembled monolayer, the alteration of the interfacial properties of the electrodes upon peptide-antibody interaction was traced by faradaic electrochemical impedance spectroscopy (EIS) using a silicotungstic heteropolyacid, H(4)SiO(4).12WO(3), as a redox probe. The electrochemical behaviour of the redox probe was evaluated with cyclic voltammetry and EIS. The effect of milk protein or 4-mercaptophenol, which was used as post-blocking agents, on the performance of the immunosensor, was investigated. Treatment with 4-mercaptophenol resulted in immunoeffective electrodes that successfully tested in anti-serum samples. An optimum dilution ratio of the samples, where the effect of the matrix on the measuring signal is negligible, was also determined.