School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, China; Shaanxi Engineering Research Center of Cardiovascular Drugs Screening & Analysis, Xi'an 710061, China.
Department of Peripheral Vascular Disease, The First Affiliated Hospital of the Medical College of Xi'an Jiaotong University, Xi'an 710061, China.
Talanta. 2024 Jan 15;267:125267. doi: 10.1016/j.talanta.2023.125267. Epub 2023 Oct 2.
A thrombin-activity-based electrochemiluminescence (ECL) biosensor was successfully constructed using tungsten oxide quantum dots (WO QD) as the co-reactant and thrombin-cleavable peptides as the recognizer. Specifically, Ru(bpy) were doped on silica nanoparticles (Ru@SiO), which greatly enhanced the ECL potential. AuNPs@WO QDs composite was then prepared to accelerate electron transfer and improve the ECL signal by 219 times. Under ideal conditions, the limit of detection for thrombin in serum was determined to be 0.28 μU/mL with a linear range from 1 μU/mL to 1 U/mL. In addition, the developed ECL biosensor was used to screen for thrombin inhibitors from 12 compounds in Artemisiae Argyi Folium. Among the compounds tested, it was observed that 100 μmol/L luteolin exhibited a significantly higher inhibition rate (exceeding 80%) compared to apigenin, isorhamnetin, naringin, or eriodictyol. In an in-vitro anticoagulation experiment, luteolin (100 μmol/L) prolonged APTT by 49%, and the molecular docking assay indicated that luteolin had binding sites of Gly219 and Asp189 in the active pockets of thrombin. This may have been the main reason underpinning luteolin's anticoagulation effects. Overall, the Ru@WO QD ECL biosensor provided a reliable strategy for thrombin activity assay and screening of anticoagulant agents.
一种基于凝血酶活性的电化学发光（ECL）生物传感器成功构建，该传感器以氧化钨量子点（WO QD）作为共反应物，以凝血酶可切割肽作为识别物。具体来说，将 Ru(bpy)掺杂到硅纳米颗粒（Ru@SiO）中，这大大增强了 ECL 势。然后制备了 AuNPs@WO QDs 复合材料，通过加速电子转移，使 ECL 信号增强了 219 倍。在理想条件下，血清中凝血酶的检测限为 0.28 μU/mL，线性范围为 1 μU/mL 至 1 U/mL。此外，所开发的 ECL 生物传感器用于从艾叶中筛选 12 种化合物的凝血酶抑制剂。在所测试的化合物中，发现 100 μmol/L 木犀草素的抑制率（超过 80%）明显高于芹菜素、异鼠李素、柚皮苷或圣草酚。在体外抗凝实验中，木犀草素（100 μmol/L）将 APTT 延长了 49%，分子对接实验表明，木犀草素在凝血酶的活性口袋中与 Gly219 和 Asp189 具有结合位点。这可能是木犀草素抗凝作用的主要原因。总体而言，Ru@WO QD ECL 生物传感器为凝血酶活性检测和抗凝剂筛选提供了可靠的策略。
