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用于通过“开启”反向自猝灭荧光检测癌细胞的适配体共轭聚合物纳米颗粒。

Aptamer-conjugated polymeric nanoparticles for the detection of cancer cells through "turn-on" retro-self-quenched fluorescence.

作者信息

Ho Lin-Chen, Wu Wei-Cheng, Chang Chang-Yu, Hsieh Hao-Hsuan, Lee Ching-Hsiao, Chang Huan-Tsung

机构信息

†Department of Chemistry, National Taiwan University, 1, Section 4, Roosevelt Road, Taipei, Taiwan.

‡Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan and National Tsing-Hua University, Hsinchu, Taiwan.

出版信息

Anal Chem. 2015;87(9):4925-32. doi: 10.1021/acs.analchem.5b00569. Epub 2015 Apr 21.

Abstract

We have developed a simple, sensitive, and rapid fluorescence assay for the detection of cancer cells, based on "turn-on" retro-self-quenched fluorescence inside the cells. 1,3-Phenylenediamine resin (DAR) nanoparticles (NPs) containing rhodamine 6G (R6G) are conjugated with aptamer (apt) sgc8c to prepare sgc8c-R6GDAR NPs, while that containing rhodamine 101 (R101) are conjugated with TD05 for the preparation of TD05-R101DAR NPs. The sgc8c-R6GDAR and TD05-R101DAR NPs separately recognize CCRF-CEM and Ramos cells. The fluorescence intensities of the two apt-DAR NPs are both weak due to self-quenching, but they increase inside the cells as a result of release of the fluorophores from the apt-DAR NPs. The apt-DAR NPs' structure becomes less compact at low pH value, leading to the release of the fluorophores. The sgc8c-R6GDAR and TD05-R101DAR NPs allow detection of as low as 44 CCRF-CEM cells and 79 Ramos cells mL(-1), respectively, using a commercial reader within 10 min. Practicality of the two probes have been validated by the quantitation and identification of CCRF-CEM and Ramos cells spiked in blood samples through conventional fluorescence and flow cytometry analysis, with advantages of sensitivity, selectivity, and rapidity.

摘要

我们基于细胞内“开启”的反向自猝灭荧光,开发了一种用于检测癌细胞的简单、灵敏且快速的荧光检测方法。含有罗丹明6G(R6G)的1,3 - 苯二胺树脂(DAR)纳米颗粒(NPs)与适配体(apt)sgc8c偶联,制备sgc8c - R6GDAR NPs,而含有罗丹明101(R101)的则与TD05偶联,用于制备TD05 - R101DAR NPs。sgc8c - R6GDAR和TD05 - R101DAR NPs分别识别CCRF - CEM和Ramos细胞。由于自猝灭,两种适配体 - DAR NPs的荧光强度都较弱,但由于荧光团从适配体 - DAR NPs中释放,它们在细胞内的荧光强度会增加。在低pH值下,适配体 - DAR NPs的结构变得不那么紧密,导致荧光团释放。使用商用读数仪,sgc8c - R6GDAR和TD05 - R101DAR NPs分别能够在10分钟内检测低至44个CCRF - CEM细胞和79个Ramos细胞mL(-1)。通过传统荧光和流式细胞术分析对掺入血样中的CCRF - CEM和Ramos细胞进行定量和鉴定,验证了这两种探针的实用性,具有灵敏度高(原文sensitivity误写成sensitity)、选择性好和快速的优点。

需注意,原文中sensitity拼写错误,正确拼写为sensitivity 。

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