Cascaded signal amplification via target-triggered formation of aptazyme for sensitive electrochemical detection of ATP
Cascaded signal amplification via target-triggered formation of aptazyme for sensitive electrochemical detection of ATP
复制标题
通过目标触发适体酶的形成进行级联信号放大，用于 ATP 的灵敏电化学检测
DOI：
10.1016/j.bios.2017.11.005
复制
发表时间：
2018-04-15
影响因子：
12.6
通讯作者：
Xiang, Yun
中科院分区：
文献类型：
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作者：
Li, Xia;Yang, Jianmei;Xiang, Yun
由于三磷酸腺苷（ATP）在各种酶活性和生物过程中的重要作用，构建可靠的ATP传感器越来越受到人们的关注。基于一个级联的，显着的信号放大的方法，通过整合的适体酶和催化发夹组件（CHA），我们已经开发了一个敏感的电化学传感器检测ATP。靶ATP导致适体酶序列的构象变化及其与发夹底物的缔合以形成活性适体酶，其中发夹底物在缓冲液中被金属离子辅因子循环切割以释放酶序列，所述酶序列也可以结合发夹底物以产生活性DNA酶。因此，适体酶/DNA酶中发夹底物的催化裂解导致产生大量中间序列。随后，这些中间序列通过CHA触发电极表面上许多亚甲蓝标记的信号序列的催化捕获，产生显著放大的电流响应，用于在0.6 nM下灵敏地检测ATP。此外，开发的传感器可以区分ATP从类似的干扰分子，并被应用于人血清样品，使传感器的一个有用的除了竞技场的小分子的灵敏检测。
The construction of reliable sensors for adenosine triphosphate (ATP) detection gains increasing interest because of its important roles in various enzymatic activities and biological processes. Based on a cascaded, significant signal amplification approach by the integration of the aptazymes and catalytic hairpin assembly (CHA), we have developed a sensitive electrochemical sensor for the detection of ATP. The target ATP leads to the conformational change of the aptazyme sequences and their association with the hairpin substrates to form active aptazymes, in which the hairpin substrates are cyclically cleaved by the metal ion cofactors in buffer to release the enzymatic sequences that can also bind the hairpin substrates to generate active DNAzymes. The catalytic cleavage of the hairpin substrates in the aptazymes/DNAzymes thus results in the generation of a large number of intermediate sequences. Subsequently, these intermediate sequences trigger catalytic capture of many methylene blue-tagged signal sequences on the electrode surface through CHA, producing significantly amplified current response for sensitive detection of ATP at 0.6 nM. Besides, the developed sensor can discriminate ATP from analogous interference molecules and be applied to human serum samples, making the sensor a useful addition to the arena for sensitive detection of small molecules.