DNA oligonucleotides as receptor layers of electrochemical sensors

The need for elaboration of analytical devices of small dimensions and the accessibility of novel nanomaterials caused the increase in the number of publications referring to the development of biosensors. DNA-based biosensors are of special interest and they were primarily used for the determination of a specific sequence which is crucial in the detection of cancer, genetic mutations, pathogens, as well as analysis of modified food. Interestingly, they could be also applied for the detection of other analytes including heavy metal ions, especially in connection with electrochemical techniques. It should be noted that the design of DNA biosensor concerns not only the development of transducer, but also careful preparation of sensing layer and the choice of the method of analytical signal generation. Selectivity is one of the essential parameter of the biosensor that determines its utility, particularly in real samples of complex matrices. In case of DNA sensors dedicated for the detection of complementary sequence, high selectivity is provided by the hybridization process. A pronounced specificity of sensing layer-analyte interaction can be also achieved with the use of functional nucleic acids - aptamers, which change their conformation upon binding an analyte. Herein, DNA-modified electrodes were firstly used for the detection of uranyl ions, as they exhibit high affinity towards phosphate moieties of nuclec acids. It was shown that UO2 2+ interacts with sensing layer independently from the chosen oligonucleotide sequence. Moreover, the influence of Pb2+ was reduced by elimination of adenine, which strongly interacts with lead ions. Another oligonucleotide-based sensor was developed for detection of mercury ions. The results indicate that Hg2+ concentration can be determined only with the use of sequence containing 100% thymine residues. Oligonucleotide-based sensor with receptor layer containing aptamers was elaborated for the detection of Pb2+ ions. In the presence of lead cations, an aptamer probe forms a G-quadruplex structure, a proposed biosensor could be characterized with selectivity towards Pb2+ The performance of DNA-based sensors for UO2 2+, Hg2+ and Pb2+ ions was optimized and addressed the choice of the manner of analytical signal generation, the influence of electrode modification with blocking agent, sensitivity dependence on the oligonucleotide sequence and the possibility of regeneration of sensing layer. Finally, the utility of proposed DNA sensors was tested by analysis in real samples