School of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, China.
School of Pharmacy, Shandong Technology Innovation Center of Molecular Targeting and Intelligent Diagnosis and Treatment, Binzhou Medical University, Yantai, 264003, China; Yantai Affiliated Hospital of Binzhou Medical University, Yantai, 264100, China.
Talanta. 2024 Jan 15;267:125149. doi: 10.1016/j.talanta.2023.125149. Epub 2023 Sep 2.
Nicotinamide adenine dinucleotide (NADH) plays a pivotal role in metabolism. Convenient detection of NADH and its related metabolites has the pursuit of point-of-care and clinical analysis. Here, we propose a polymer dots (Pdots)-based NADH-sensitive electrochemiluminescence (ECL) biosensor for detection of NADH and three metabolites. Pdots acted as the efficient ECL emitters without additional modification to construct this biosensor. Specially, NADH both acted as the final detection target and at the same time as the bio-coreactants to sensitively influence the ECL intensities, in which NADH was generated or consumed in the presence of the target analyte and their specific enzyme. For glucose and lactic acid detection, NAD was reduced to NADH to generate an enhanced ECL signal. Conversely, for pyruvate detection, NADH was consumed to further decrease the ECL. The designed Pdots-based ECL biosensor showed wide detection ranges, high selectivity and low limits of detection of 4.6 μM, 0.7 μM and 0.5 μM for the analysis of three analytes, respectively. This strategy was successfully applied in quantifying the concentrations of glucose, lactic acid and pyruvate in human serum, which also has the potential to be implemented as a powerful and fast tool for ECL sensing of NADH and other related metabolites for point-of-care use and disease monitoring.
烟酰胺腺嘌呤二核苷酸(NADH)在新陈代谢中起着关键作用。方便地检测 NADH 及其相关代谢物一直是即时检测和临床分析的追求。在这里,我们提出了一种基于聚合物点(Pdots)的 NADH 敏感电化学发光(ECL)生物传感器,用于检测 NADH 和三种代谢物。Pdots 无需额外修饰即可作为高效的 ECL 发射器来构建这种生物传感器。特别是,NADH 既可以作为最终的检测目标,同时也可以作为生物反应试剂,以敏感地影响 ECL 强度,其中在存在目标分析物及其特定酶的情况下,NADH 被生成或消耗。对于葡萄糖和乳酸检测,NAD 被还原为 NADH 以产生增强的 ECL 信号。相反,对于丙酮酸检测,NADH 被消耗以进一步降低 ECL。设计的基于 Pdots 的 ECL 生物传感器显示出宽的检测范围、高选择性和低检测限,分别为 4.6μM、0.7μM 和 0.5μM,用于分析三种分析物。该策略成功应用于定量测定人血清中葡萄糖、乳酸和丙酮酸的浓度,也有望作为即时检测 NADH 和其他相关代谢物的强大、快速的 ECL 传感工具,用于即时检测和疾病监测。