A new fluorescence "switch on" assay for heparin detection by using a functional ruthenium polypyridyl complex.
A new fluorescence "switch on" assay for heparin detection by using a functional ruthenium polypyridyl complex.
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DOI：
10.1039/c3an00242j
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发表时间：
2013-05
期刊：
影响因子：
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通讯作者：
Ting Cheng;Junliang Yao;Xing Gao;Wenliang Sun;Shuo Shi;Tianming Yao
中科院分区：
文献类型：
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作者：
Ting Cheng;Junliang Yao;Xing Gao;Wenliang Sun;Shuo Shi;Tianming Yao
在本研究中，通过监测功能性钌多吡啶配合物([Ru(phen)（2)dppz-idzo](2+)，配合物1）在缓冲溶液中的释放变化，建立了一种新的肝素检测和定量策略，如胎牛血清（FBS）。发现聚阴离子肝素通过静电效应和/或氢键相互作用与带正电的ru -络合物相互作用，从而导致ru -络合物的显著荧光增强。为了深入了解这种荧光“开启”行为，基于荧光和紫外吸收结果，通过分子对接模拟，建立了ru -配合物与肝素的结合模型。荧光分析的选择性结果表明，我们的配合物对肝素的荧光选择性优于其类似物，如硫酸软骨素（Chs）或透明质酸（Hya），它们与肝素相比具有较低的电荷密度和/或结构相容性。对稀释的FBS溶液中肝素进行定量，在0.01 ~ 4.87 U mL（-1）范围内观察到线性校准曲线（检出限为0.01 U mL(-1)）。这种“一步”荧光“开启”法检测肝素，无标记、方便、灵敏、选择性强，发射波长长，斯托克斯位移大。
In the present study, a new strategy for heparin detection and quantification in biological media, such as fetal bovine serum (FBS), is developed by monitoring the emission change of a functional ruthenium polypyridyl complex ([Ru(phen)(2)dppz-idzo](2+), complex 1) in buffer solution. Polyanionic heparin is found to interact with a positively charged Ru-complex through electrostatic effects and/or hydrogen bonding interactions, which leads to a significant fluorescence enhancement of the Ru-complex. To get insight into this fluorescence "switch on" behavior, the binding model of the Ru-complex to heparin is established by employing molecular docking simulations based on the fluorescence and UV absorption results. The selectivity results of the fluorescence assay reveal that our complex displayed good fluorescence selectivity towards heparin over its analogues, such as chondroitin 4-sulfate (Chs) or hyaluronic acid (Hya), which have lower charge density and/or structural compatibility as compared to that of heparin. Quantification of heparin is also performed and a linear calibration curve is observed in the range of 0.01-4.87 U mL(-1) (the limit of detection is 0.01 U mL(-1)) for heparin detection in diluted FBS solution. This "one-step" fluorescence "switch on" assay for heparin detection is label-free, convenient, sensitive and selective, and has a long emission wavelength and large Stokes shift.