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可活化发射激基复合物的形成用于高选择性检测β-半乳糖苷酶。

Activatable Formation of Emissive Excimers for Highly Selective Detection of β-Galactosidase.

机构信息

Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, Key Lab of Modern Separation Science in Shaanxi Province, College of Chemistry and Materials Science, Northwest University, Xi'an, Shaanxi 710127, People's Republic of China.

Shaanxi Provincal Key Laboratory of Papermaking Technology and Specialty Paper Development, College of Bioresources Chemical and Materials Engineering, Shaanxi University of Science and Technology, Xi'an, Shaanxi 710021, People's Republic of China.

出版信息

Anal Chem. 2020 Apr 21;92(8):5733-5740. doi: 10.1021/acs.analchem.9b04806. Epub 2020 Mar 31.

Abstract

Small-molecular fluorescence sensors have become promising detection tools in many fields attributing to their high sensitivity, excellent temporal and spatial resolution, and low cytotoxicity. However, high concentration or aggregation-induced fluorescence quenching effect has usually hindered the development of traditional fluorescence dyes. Herein, a new fluorophore cyanovinylene dye with excimer emission property has been constructed. It shows an obvious enhanced and red-shift emission upon aggregation in aqueous solution, which overmatches the conventional pyrene with longer absorption and emission wavelengths. Using this unique optical property, a new fluorescence probe has been developed for trapping of β-galactosidase (β-Gal) activity with high selectivity, low limit of detection of 0.17 U, and rapid recognition, which is based on the β-Gal-induced formation of red-shift emitting excimer. β-Gal has a strong affinity for , which is verified through the Michaelis-Menten constants (, 1.87 μM) and the hydrolysis efficiencies (/, 1.78 × 10 M s). Furthermore, the assay system has been successfully used for detecting endogenous β-Gal in living ovarian cancer cells and can passively targeted to identify β-Gal in organelle level and determine its subcellular location with satisfactory accuracy. We anticipate that the new fluorophore cyanovinylene dye herein may inaugurate new opportunities for the development of excimer emission sensors.

摘要

小分子荧光传感器由于其高灵敏度、优异的时空分辨率和低细胞毒性,已成为许多领域有前途的检测工具。然而,高浓度或聚集诱导的荧光猝灭效应通常阻碍了传统荧光染料的发展。本文构建了一种具有激基缔合物发射特性的新型荧光团氰基乙烯染料。它在水溶液中聚集时表现出明显增强的红移发射,这超过了传统的蒽,具有更长的吸收和发射波长。利用这一独特的光学性质,开发了一种新的荧光探针,用于β-半乳糖苷酶(β-Gal)活性的高选择性捕获,检测限低至 0.17 U,且具有快速识别能力,这是基于β-Gal 诱导形成的红移发射激基缔合物。β-Gal 与 具有很强的亲和力,这通过米氏常数(,1.87 μM)和水解效率(/,1.78×10 M s)得到验证。此外,该测定系统已成功用于检测活卵巢癌细胞中的内源性β-Gal,并可被动靶向识别细胞器水平的β-Gal,确定其亚细胞定位,具有令人满意的准确性。我们预计本文中的新型荧光团氰基乙烯染料可能为激基缔合物发射传感器的发展开辟新的机会。

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