Wang Zhi-Gang, Yan Xiao-Jing, Liu Hai-Bo, Zhang De-Long, Liu Wei, Xie Cheng-Zhi, Li Qing-Zhong, Xu Jing-Yuan
Department of Chemical Biology and Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, P. R. China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100193, P. R. China.
J Mater Chem B. 2020 Sep 23;8(36):8346-8355. doi: 10.1039/d0tb01411g.
Human serum albumin (HSA) is considered as a biomarker for the early diagnosis of renal disease, therefore identifying and detecting HSA in biological fluids (especially urine) with an easy method is of great importance. Herein, we report a novel hydrazide Schiff base fluorescent probe N'-((7-(diethylamino)-2-oxo-2H-chromen-3-yl)methylene)pyrazine-2-carbohydrazide (NPC), which self-assembled into nanoparticles in aqueous solution. Based on disassembly-induced emission and the site-specific recognition mechanism, the binding of NPC with HSA resulted in a fluorescence "turn-on" response. Probe NPC exhibited superior selectivity and sensitivity toward HSA with a detection limit of 0.59 mg L-1 in PBS and 0.56 mg L-1 in the urine sample. The site-binding mechanism of NPC with HSA was explored by fluorescence quenching study, Job's plot analysis, HSA destruction, site marker displacement and molecular docking. Fluorescence imaging of HSA in MCF-7 cells was achieved by using a non-toxic NPC probe, suggesting that NPC could be applied to visualize the level of HSA in vivo. More importantly, further practical applications of probe NPC in human urine samples were achieved with satisfactory results by using a fluorometer or test paper, which could provide extensive application in clinical diagnosis.
人血清白蛋白(HSA)被认为是肾脏疾病早期诊断的生物标志物,因此用简便方法在生物流体(尤其是尿液)中识别和检测HSA具有重要意义。在此,我们报道了一种新型的酰肼席夫碱荧光探针N'-((7-(二乙氨基)-2-氧代-2H-色烯-3-基)亚甲基)吡嗪-2-碳酰肼(NPC),它在水溶液中自组装成纳米颗粒。基于解组装诱导发光和位点特异性识别机制,NPC与HSA的结合导致荧光“开启”响应。探针NPC对HSA表现出优异的选择性和灵敏度,在PBS中的检测限为0.59 mg L-1,在尿液样本中的检测限为0.56 mg L-1。通过荧光猝灭研究、Job曲线分析、HSA破坏、位点标记置换和分子对接等方法探究了NPC与HSA的位点结合机制。使用无毒的NPC探针实现了MCF-7细胞中HSA的荧光成像,表明NPC可用于可视化体内HSA水平。更重要的是,通过使用荧光计或试纸,探针NPC在人尿液样本中实现了进一步的实际应用,结果令人满意,这可为临床诊断提供广泛应用。