Electrochemical determination of the activity and inhibition of telomerase based on the interaction of DNA with molybdate.

Abstract

An ultrasensitive electrochemical sensor is described for the determination of the activity of telomerase. It is based on a DNA-generated current that is due to the reaction of the phosphate groups on DNA with molybdate to form a redox-active molybdophosphate. A telomerase substrate primer was first immobilized on a gold electrode. In the presence of telomerase and deoxyribonucleoside triphosphates (dNTPs), the primer can be extended with repetitive nucleotide sequences (TTAGGG). The subsequent reaction of the sensor with molybdate results in the enhancement of electrochemical current intensity due to an increased amount of nucleotides on the electrode. Sensitivity can be further improved by introducing a hairpin probe that partially hybridizes with the repetitive TTAGGG sequence and further enhances the amount of DNA on the electrode. The biosensor, best operated at 0.2V (vs. Ag/AgCl) shows a linear response to telomerase activity from 1×102 to 107 Hela cells mL-1. The assay was applied to the detection of telomerase activity in HeLa cancer cells treated with the anticancer drug epigallocatechin gallate, and the results indicate that it holds great potential in anticancer drug screening. Graphical abstract Schematic presentation of anultrasensitive electrochemical sensor for the determination of telomerase activity based on DNA generated electrochemical current. dNTPs in the scheme represents deoxyribonucleoside triphosphates.