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首页> 外文期刊>Chemistry: A European journal >Towards Ratiometric Sensing of Amyloid Fibrils In Vitro
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Towards Ratiometric Sensing of Amyloid Fibrils In Vitro

机译：走向淀粉样原纤维的比例传感

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The aggregation of amyloid-beta peptide and its accumulation in the human brain has an important role in the etiology of Alzheimer's disease. Thioflavin T has been widely used as a fluorescent marker for these amyloid aggregates. Nevertheless, its complex photophysical behavior, with strong wavelength dependencies of all its fluorescence properties, requires searching for new fluorescent probes. The use of 2-(2'-hydroxyphenyl)imidazo[4,5-b]pyridine (HPIP), which shows two emission bands and a rich excited-state behavior due to the existence of excited-state intramolecular processes of proton transfer and charge transfer, is proposed. These properties result in a high sensitivity of HPIP fluorescence to its microenvironment and cause a large differential fluorescence enhancement of the two bands upon binding to aggregates of the amyloid-beta peptide. Based on this behavior, a very sensitive ratiometric method is established for the detection and quantification of amyloid fibrils, which can be combined with the monitoring of fluorescence anisotropy. The binding selectivity of HPIP is discussed on the basis of the apparent binding equilibrium constants of this probe to amyloid-beta (1-42) fibrils and to the nonfibrillar protein bovine serum albumin. Finally, an exhaustive comparison between HPIP and thioflavin T is presented to discuss the sensitivity and specificity of these probes to amyloid aggregates and the significant advantages of the HPIP dye for quantitative determinations.
机译：β淀粉样蛋白肽的聚集及其在人脑中的积累在阿尔茨海默氏病的病因学中具有重要作用。硫黄素T已被广泛用作这些淀粉状蛋白聚集体的荧光标记。然而，其复杂的光物理行为以及所有荧光特性的强烈波长依赖性，需要寻找新的荧光探针。使用2-（2'-羟基苯基）咪唑并[4,5-b]吡啶（HPIP），由于存在质子转移和分子的激发态分子内过程，因此显示了两个发射带和丰富的激发态行为。建议进行电荷转移。这些特性导致HPIP荧光对其微环境具有高度敏感性，并在与淀粉样β肽聚集体结合后导致两个条带的荧光差异较大。基于此行为，建立了一种非常灵敏的比率测定方法来检测和定量淀粉样蛋白原纤维，该方法可以与荧光各向异性的监测相结合。基于该探针对淀粉样β（1-42）纤维和对非原纤维蛋白牛血清白蛋白的表观结合平衡常数，讨论了HPIP的结合选择性。最后，提出了HPIP和硫黄素T之间的详尽比较，以讨论这些探针对淀粉样蛋白聚集体的敏感性和特异性，以及HPIP染料在定量测定中的显着优势。

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