Photoelectrochemical aptasensing for thrombin based on exonuclease III-assisted recycling signal amplification and nanoceria enzymatic strategy.

In the present work, a capture DNA (c-DNA) was immobilized on the TNA/g-CN to develop a sensitive and selective TNA/g-CN/c-DNA photoelectrochemical aptasensor for determining thrombin. With the aid of the specific recognition of anti-thrombin aptamer towards thrombin, ingenious design of hairpin DNA, and exonuclease III-assisted recycling signal amplification, more nanoceria could be assembled on the TNA/g-CN/c-DNA to form TNA/g-CN/nanoceria in the presence of thrombin. Due to the oxidase-mimic catalytic efficiency of nanoceria and the oxygen consumption for glucose oxidation, the photoexcited electrons at the conduction band of g-CN could be well transferred to that of TNA under visible light irradiation, resulting in the increase of the photocurrent of TNA/g-CN/nanoceria, and the increase value of photocurrent had a linear relationship with the concentration of thrombin under the optimal conditions. So, the constructed TNA/g-CN/c-DNA photoelectrochemical aptasensor exhibited a satisfactory quantitative range from 0.01 pM to 0.5 nM, low detection limit with 3.4 fM for thrombin determination, and was applied for the human serum analysis successfully with RSD of less than 4.8% and the recovery between 95% and 113%.