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Ultrasensitive determination of thrombin by using an electrode modified with WSe2 and gold nanoparticles, aptamer-thrombin-aptamer sandwiching, redox cycling, and signal enhancement by alkaline phosphatase

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Abstract

A sensitive aptamer/protein binding-triggered sandwich assay for thrombin is described. It is based on electrochemical-chemical-chemical redox cycling using a glassy carbon electrode (GCE) that was modified with WSe2 and gold nanoparticles (AuNPs). The AuNPs are linked to thrombin aptamer 1 via Au-S bonds. Thrombin is first captured by aptamer 1 and then sandwiched through the simultaneous interaction with AuNPs modified with thrombin-specific aptamer 2 and signalling probe. Subsequently, the DNA-linked AuNP hybrids result in the capture of streptavidin-conjugated alkaline phosphatase onto the modified GCE through the specific affinity reaction for further signal enhancement. As a result, a linear range of 0–1 ng mL−1 and a detection limit as low as 190 fg mL−1 are accomplished. The specificity for thrombin is excellent. Conceivably, this strategy can be easily expanded to other proteins by using the appropriate aptamer.

Schematic presentation of an electrochemical biosensor for thrombin based on WSe2 and gold nanoparticles, aptamer-thrombin-aptamer sandwiching, redox cycling, and signal enhancement by alkaline phosphatase.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (21475115), Henan Provincial Science and technology innovation team (C20150026), Nanhu Scholars Program of XYNU and Henan Science and Technology Cooperation Project (172106000064).

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Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang, 464000, China

    Yi-Han Wang, Huan Xia, Ke-Jing Huang, Ying-Ying Ma, Rui Deng, Yun-Fei Lu & Zi-Wei Han

  2. School of Physics and Electronic Engineering, Xinyang Normal University, Xinyang, 464000, China

    Xu Wu

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  1. Yi-Han Wang
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Correspondence to Ke-Jing Huang.

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Wang, YH., Xia, H., Huang, KJ. et al. Ultrasensitive determination of thrombin by using an electrode modified with WSe2 and gold nanoparticles, aptamer-thrombin-aptamer sandwiching, redox cycling, and signal enhancement by alkaline phosphatase. Microchim Acta 185, 502 (2018). https://doi.org/10.1007/s00604-018-3028-7

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