基于荧光共振能量转移的开启型适配体传感器用于肌酸激酶同工酶（CK-MB）的灵敏检测

FRET-Based TURN-ON Aptasensor for the Sensitive Detection of CK-MB.

作者信息

Asghar Rabia, Rasheed Madiha, Lv Xuefei, Deng Yulin

机构信息

Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, School of Medical Technology, Beijing Institute of Technology, Beijing 100081, China.

BIT & GS Technology Co., Ltd., Beijing 100043, China.

出版信息

Biosensors (Basel). 2025 Jul 11;15(7):446. doi: 10.3390/bios15070446.

A fluorescent sandwich assay was devised to quantify CK-MB. In a typical immunoassay, antibodies bind to the target, and the detected signal is quantified according to the target's concentration. We innovated a unique fluorescence assay known as the "enzyme-linked aptamer assay" (ELAA) by substituting antibodies with a pair of high-affinity aptamers labelled with biotin, namely apt. A1 and apt. A2. Avidin-labelled ALP binds to biotin-labelled aptamers, hydrolyzing its substrate, 2-phosphoascorbic acid trisodium salt, resulting in the formation of ascorbic acid. The catalytic hydrolysate functions as a reducing agent, causing the deterioration of MoS nanosheets. This results in the transformation of MoS nanosheets into nanoribbons, leading to the release of quenched AGQDs. The reestablishment of fluorescence is triggered by Förster Resonance Energy Transfer (FRET) between the MoS nanoribbons and AGQDs, enhancing the sensitivity of disease biomarker detection. The working range for detection falls between 2.5 nM and 160 nM, and the limit of detection (LOD) for CK-MB is verified at 0.20 nM.

设计了一种荧光夹心测定法来定量肌酸激酶同工酶（CK-MB）。在典型的免疫测定中，抗体与靶标结合，然后根据靶标的浓度对检测信号进行定量。我们创新了一种独特的荧光测定法，称为“酶联适配体测定法”（ELAA），用一对标记有生物素的高亲和力适配体（即适配体A1和适配体A2）替代抗体。抗生物素蛋白标记的碱性磷酸酶（ALP）与生物素标记的适配体结合，水解其底物2-磷酸抗坏血酸钠盐，生成抗坏血酸。催化水解产物作为还原剂，导致二硫化钼（MoS）纳米片变质。这导致MoS纳米片转变为纳米带，从而使猝灭的聚集诱导发光量子点（AGQDs）释放出来。二硫化钼纳米带与AGQDs之间的荧光共振能量转移（FRET）触发荧光的重新建立，提高了疾病生物标志物检测的灵敏度。检测的工作范围在2.5 nM至160 nM之间，CK-MB的检测限（LOD）经验证为0.20 nM。