Sub-picomolar label-free detection of thrombin using electrochemical impedance spectroscopy of aptamer-functionalized...

An ultrasensitive aptasensor for the label free non-faradaic detection of thrombin has been demonstrated on molybdenum disulphide (MoS₂) nanosheets. These nanosheets were physiochemically immobilized onto a silicon micro-electrode platform. Thrombin detection was achieved through the charge modulation of the electrical double layer due to the specific and dose dependent binding of thrombin to the surface of thiol terminated ssDNA aptamer functionalized MoS₂ nanosheets. Electrical double layer charge modulation associated with thrombin binding was characterized using electrochemical impedance spectroscopy. Dynamic light scattering was also used to confirm the dose dependent behavior. ATR-FTIR spectroscopy and XPS analysis were independently used to validate the functionalization of the ssDNA aptamer onto MoS₂ nanosheets. ssDNA aptamer functionalized molybdenum disulfide (MoS₂) for selective and specific capture of thrombin was demonstrated both in phosphate buffered saline (PBS) and human serum. The optimized immunoassay enabled the detection of thrombin ranging from 267 fM to 267 pM in phosphate buffer. The limit of detection of 53 pM and the linear dynamic range of detection of thrombin ranged from 53 to 854 pM in human serum. The rapid response time for the electrochemical impedance spectroscopy signal makes it an attractive option for the real-time detection of thrombin based point-of-care diagnostic devices.