Abstract
Thrombin, a crucial enzyme involved in blood coagulation and associated diseases, requires accurate detection of its activity and screening of inhibitors for clinical diagnosis and drug discovery. To address this, an electrochemiluminescence (ECL) method was developed to detect thrombin activity based on the sensitization of Ti3C2Tx MXene, which could sensitize the Ru(bpy)32+ ECL system greatly. The thrombin-cleavable substrate bio-S-G-R-P–V-L-G-C was used as recognizer to evaluate the activity of thrombin. Under the optimal conditions, the limit of detection for thrombin in serum was 83 pU/mL (S/N = 3) with a linear range from 0.1 nU/mL to 1 µU/mL. Moreover, the developed ECL biosensor was employed to screen for thrombin inhibitors from Artemisiae argyi Folium. Four potential thrombin inhibitors (isoquercitrin, nepetin, L-camphor, L-borneol) were screened out with inhibition rates beyond 50%, among which isoquercitrin had the best inhibition rate of 90.26%. Isoquercitrin and nepetin were found to be competitive inhibitors of thrombin, with \({K}_{i}^{app}\) values of 0.91 μM and 2.18 μM, respectively. Molecular docking results showed that these compounds could interact with the active sites of thrombin through hydrogen bonds including ASP189, SER195, GLY216, and GLY219. The electrochemical biosensor constructed provides a new idea for the detection of thrombin activity and screening of its inhibitors.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Financial support from the National Natural Science Foundation of China (81973277, 82273891, 82003709, 22274126) and the National Natural Science Foundation of Shaanxi Province (2021JM-039, 2023-YBSF-262) are gratefully acknowledged.
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Deng, Z., Tan, X., Guo, D. et al. MXene-sensitized electrochemiluminescence sensor for thrombin activity detection and inhibitor screening. Microchim Acta 190, 328 (2023). https://doi.org/10.1007/s00604-023-05906-9
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DOI: https://doi.org/10.1007/s00604-023-05906-9