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一种基于碳纳米管的荧光适配体传感器,用于高灵敏度检测腺苷脱氨酶活性及天然提取物中的抑制剂筛选。

A carbon nanotubes based fluorescent aptasensor for highly sensitive detection of adenosine deaminase activity and inhibitor screening in natural extracts.

作者信息

Hu Kun, Huang Yong, Wang Sheng'e, Zhao Shulin

机构信息

Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China.

Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), College of Chemistry and Pharmacy, Guangxi Normal University, Guilin 541004, China.

出版信息

J Pharm Biomed Anal. 2014 Jul;95:164-8. doi: 10.1016/j.jpba.2014.02.027. Epub 2014 Mar 12.

Abstract

A carbon nanotubes (CNTs) based fluorescent aptasensor was developed for adenosine deaminase (ADA) activity detection and inhibitor screening by using adenosine (AD) as the substrate. This sensing system consists of CNTs, AD, split anti-AD aptamer fragment and dye-labeled aptamer fragment. In the absence of ADA, two aptamer fragments bind simultaneously with AD to form an AD-aptamer complex. This AD-aptamer complex cannot adsorb onto CNTs, and has high fluorescence intensity. When ADA is introduced into this system, ADA can convert AD into inosine, which has not affinity to the split anti-AD aptamer fragment. Thus, the split anti-AD aptamer fragments were adsorbed onto CNTs via strong π-π stacking interactions, resulting in the quenching of the fluorescence of the dye-labeled aptamer fragment. The proposed aptasensor can detect ADA activity from 0.005 to 0.2U/mL with a low detection limit of 0.002 U/mL. Moreover, it has been also demonstrated that this CNTs-based fluorescence aptasensor is suitable for ADA inhibitor screening from traditional Chinese medicine (TCM). Considering the superior sensitivity and specificity, the proposed CNTs-based fluorescent aptasensor can be expected to provide a simple, cost-effective and sensitive platform for the detection of ADA activity and screening of potential drugs.

摘要

基于碳纳米管(CNTs)构建了一种荧光适配体传感器,用于以腺苷(AD)为底物检测腺苷脱氨酶(ADA)活性并筛选抑制剂。该传感系统由碳纳米管、腺苷、抗腺苷适配体片段和染料标记的适配体片段组成。在不存在ADA的情况下,两个适配体片段与腺苷同时结合形成腺苷 - 适配体复合物。这种腺苷 - 适配体复合物不能吸附在碳纳米管上,具有高荧光强度。当将ADA引入该系统时,ADA可将腺苷转化为次黄嘌呤核苷,其对拆分的抗腺苷适配体片段没有亲和力。因此,拆分的抗腺苷适配体片段通过强π-π堆积相互作用吸附在碳纳米管上,导致染料标记的适配体片段荧光猝灭。所提出的适配体传感器能够检测0.005至0.2U/mL的ADA活性,检测限低至0.002U/mL。此外,还证明了这种基于碳纳米管的荧光适配体传感器适用于从中药中筛选ADA抑制剂。考虑到其卓越的灵敏度和特异性,所提出的基于碳纳米管的荧光适配体传感器有望为ADA活性检测和潜在药物筛选提供一个简单、经济高效且灵敏的平台。

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