A Label-free Resonance Rayleigh Scattering Sensor for Detection of Thrombin Based on Aptamer Recognizing

Abstract

The interaction between thrombin binding aptamer (TBA) and thrombin (TB) was studied by resonance Rayleigh scattering (RRS). In neutral medium, TBA is present in a balanced form between a G-quadruplex structure and a random coil structure, and the TBA can be induced by metal ions to form a G-quadruplex structure. Upon addition of thrombin, the G-quadruplex selectively bound to TB, which resulted in enhanced resonance Rayleigh scattering. The scattering intensities increased proportionally with the concentration of TB from 10 to 50 nM. The method had very high sensitivity and good selectivity, and the detection limit (3δ/s) was 1 nM. In this work, the spectral characteristics of RRS, the optimum conditions of the reaction, and influencing factors for the RRS intensities were investigated. Furthermore, the structure of the TBA-TB complex and the sensing mechanism were explored. The TB sensor was applied to a diluted human serum sample with satisfactory results, indicating the potential of this method to be applied to biological samples. A selective and simple RRS sensor for the detection of trace amounts of TB is proposed based on conformational change of TBA.

Keywords: Aptamer; G-quadruplex structure; resonance Rayleigh scattering; thrombin.