Faculty of Chemistry, Bu-Ali Sina University, Hamedan, Iran.
Faculty of Chemistry, Bu-Ali Sina University, Hamedan, Iran.
Anal Chim Acta. 2020 Apr 8;1105:187-196. doi: 10.1016/j.aca.2020.01.046. Epub 2020 Jan 29.
Dopamine β-hydroxylase (DBH) is involved in various neuronal transmission processes in the brain. Due to the severe diseases caused by abnormity levels of such important enzyme in human serum, sensitive and rapid detection of DBH at early stages is crucial, particularly for clinical analysis. Herein, we developed optical sensors for DBH that include the following: (i) a ratiometric fluorescence sensor that hybridizes the bovine serum albumin (BSA)-gold nanoclusters (BSA-AuNCs) and nitrogen doped carbon dots (N-CDs). The sensor proved to be highly selective and sensitive, achieving a linear range of 0.02-0.16 μg mL and a limit of detection of 4.0 ng mL. In the presence of DBH, the fluorescence intensity of BSA-AuNCs (λ = 615 nm) was remarkably quenched by DBH serving as a reporter signal, whereas the N-CDs fluorescence intensity at 440 nm was almost kept unchanged serving as a reference signal. The developed ratiometric sensor is capable of demonstrating a color change from pink to violet and blue with a gradual increase in DBH concentration, which is discernible by the naked-eye. A test strip is prepared for semi-quantitative assay and convenient use. Intriguingly, by taking advantage of the inter-AuNCs aggregation in the presence of DBH, (ii) a resonance light scattering (RLS) sensor was also developed based on the nanohybrid probe (detection limit 95 ng mL). Fluorescence imaging in PC12 cell lines demonstrated that the BSA-AuNCs could be utilized in visualization assay towards intracellular DBH. Additionally, the sensors were tested in a real matrix by spiking serum samples with satisfactory recoveries.
多巴胺β-羟化酶（DBH）参与大脑中的各种神经元传递过程。由于人血清中这种重要酶异常水平会导致严重疾病，因此早期敏感、快速地检测 DBH 至关重要，尤其是用于临床分析。在此，我们开发了用于 DBH 的光学传感器，包括：（i）比率荧光传感器，它杂交了牛血清白蛋白（BSA）-金纳米簇（BSA-AuNCs）和氮掺杂碳点（N-CDs）。该传感器被证明具有高度选择性和灵敏度，实现了 0.02-0.16μg mL 的线性范围和 4.0ng mL 的检测限。在 DBH 的存在下，BSA-AuNCs 的荧光强度（λ=615nm）被 DBH 显著猝灭，DBH 作为报告信号，而 N-CDs 的荧光强度（λ=440nm）几乎保持不变，作为参考信号。开发的比率传感器能够显示出从粉红色到紫色和蓝色的颜色变化，随着 DBH 浓度的逐渐增加，肉眼可辨。制备测试条进行半定量测定和方便使用。有趣的是，通过利用存在 DBH 时的 AuNCs 之间的聚集，（ii）还基于纳米杂化探针开发了共振光散射（RLS）传感器（检测限 95ng mL）。在 PC12 细胞系中的荧光成像表明，BSA-AuNCs 可用于可视化细胞内 DBH 的测定。此外，通过用血清样品加标对实际基质中的传感器进行了测试，回收率令人满意。
