Effects of redox label location on the performance of an electrochemical aptamer-based tumor necrosis factor-alpha sensor.

We report the development of an electrochemical aptamer-based sensor for real time detection of tumor necrosis factor-alpha. The focus of this study is to evaluate the effects of the redox label location on the overall sensor performance, including sensor stability, detection limit, reusability, and selectivity. Three aptamer probes, each labeled with methylene blue (MB) at a specific location, were designed and employed in the fabrication of the sensors. Among the three sensors, the sensor fabricated using an aptamer with the MB label located at the distal end has a detection limit of 100 pM and is regenerable. The sensor fabricated using an aptamer with an internal MB modification has a detection limit of 10 nM and is not regenerable. Both sensors can be employed in complex biological samples such as 50% urine and 50% saliva. However, the sensor fabricated with an aptamer with the MB label located at the proximal end suffers from poor reproducibility and is highly unstable, thus limiting its application as a sensor. On the bases of these results, placing the MB label at the distal end of the aptamer probe appears to be the most advantageous for this sensor design for it does not interfere with monolayer formation and target binding.