Fluorescence coupled capillary electrophoresis as a strategy for tetrahedron DNA analysis
Fluorescence coupled capillary electrophoresis as a strategy for tetrahedron DNA analysis
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荧光耦合毛细管电泳作为四面体 DNA 分析的策略
DOI：
10.1016/j.talanta.2021.122225
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发表时间：
2021-02-24
期刊：
影响因子：
6.1
通讯作者：
Wang, Jianhao
中科院分区：
文献类型：
--
作者：
Hong, Tingting;Zheng, Ronghui;Wang, Jianhao
建立了一种基于荧光耦合毛细管电泳（CE-FL）的四面体DNA （TD）和TD-阿霉素（DOX）偶联物分析方法。毛细管凝胶电泳在分离TD和DNA链方面表现出良好的性能。在优化条件下，可获得满意的运行间重复性和日间重复性，分辨率（n = 6）的相对标准偏差值为0.64%。此外，CE和荧光检测的结合为定量TD浓度和计算TD损伤程度提供了一个灵敏的平台。电泳结果表明，CE-FL是一种特异性高、灵敏度高的TD测定方法。此外，还探讨了CE-FL在TD荧光共振能量转移（FRET）研究中的应用。两种类型的DNA链被用来干扰TD的形成。探讨了部分互补链和完全互补链对FRET信号的影响，并讨论了影响机理。在应用CE- fl表征TD之后，我们还结合CE和FRET分析了TD- dox共轭物。CE提出了一种监测DOX加载和释放过程的有利技术。这些值得注意的结果为应用CE-FL检测DNA纳米材料提供了基础。
A strategy based on fluorescence coupled capillary electrophoresis (CE-FL) was developed for analyzing tetrahedron DNA (TD) and TD-doxorubicin (DOX) conjugate. Capillary gel electrophoresis exhibited desirable performance for separating TD and DNA strands. Under the optimized conditions, satisfactory repeatability concerning run-to-run and interday repeatability was obtained, and relative standard deviation value of resolution (n = 6) was 0.64%. Furthermore, the combination of CE and fluorescence detection provided a sensitive platform for quantifying TD concentration and calculating the damage degree of TD. The electrophoretograms indicated that CE-FL was a suitable TD assay method with high specificity and sensitivity. In addition, the application of CE-FL for TD fluorescence resonance energy transfer (FRET) research was also explored. Two types of DNA strands were utilized to interfere the formation of TD. The impact of partially complementary chain and completely complementary chain on FRET signal was explored, and the influence mechanism was discussed. After applying CE-FL for characterizing TD, we also combine CE and FRET to analyze TD-DOX conjugate. CE presented a favourable technique to monitor DOX loading and releasing processes. These noteworthy results offered a stepping stone for DNA nanomaterials assay by using CE-FL.