ИСТИНА

We synthesized a series of conjugates of hemin and its aptamer EAD2, named covalent peroxidase-mimicking DNAzymes (cPMDNAzymes), varying the length, rigidity and 5’-/3’-position of the linker between the oligonucleotide and hemin. Systemic structure-activity relationship study of these cPMDNAzymes showed that cPMDNAzyme with hemin bound to the 5’-end of EAD2 via T10 spacer (cPMDNAzyme(T10)) demonstrated the highest activity in luminol oxidation assay. Its activity was significantly higher in comparison to the non-covalent complex of hemin and aptamer EAD2 (ncPMDNAzyme). Comparison of the detection limit values for the cPMDNAzyme(T10) in the reactions of oxidation of luminol and ABTS, which were equal to 0.2 and 1.6 pM, respectively, showed that the chemiluminescent method of cPMDNAzyme(T10) detection is preferred over the colorimetric one. Similarity of the detection limit values for the cPMDNAzyme(T10) and horseradish peroxidase, whose activity was measured in an enhanced chemiluminescence reaction (0.25 pM), opens up very promising perspectives for the development of highly sensitive cPMDNAzyme(T10)-based assays and devices. This work was supported by the Russian Foundation for Basic Research (13-04-00364).