Using Spectroscopy to Guide the Adaptation of Aptamers into Electrochemical Aptamer-Based Sensors.
Using Spectroscopy to Guide the Adaptation of Aptamers into Electrochemical Aptamer-Based Sensors.
复制标题
使用光谱学指导适配体适应电化学适配体传感器。
DOI：
10.1021/acs.bioconjchem.2c00275
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发表时间：
2023
影响因子：
4.7
通讯作者：
Plaxco,KevinW
中科院分区：
文献类型：
--
作者：
Wu,Yuyang;Ranallo,Simona;DelGrosso,Erica;Chamoro-Garcia,Alejandro;Ennis,HerbertL;Milosavić,Nenad;Yang,Kyungae;Kippin,Tod;Ricci,Francesco;Stojanovic,Milan;Plaxco,KevinW
基于电化学适配子(EAB)的传感器利用电极连接、氧化还原报告分子修饰的适配子结合诱导的构象变化，将目标识别转化为易于测量的电化学输出。由于这种信号转导机制是单步和快速可逆的，EAB传感器支持高频、实时的分子测量，并且由于它概括了体内自然产生的受体之间看到的无试剂、构象关联的信号，EAB传感器具有足够的选择性，可以在活体中发现的复杂、时变的环境中工作。然而，EAB传感器的制造需要对其靶标识别适配子进行修饰，以便(1)它经历必要的结合诱导的构象变化，(2)这种“构象开关”的热力学被调整，以确保它们反映了亲和力和信号增益之间的可接受的权衡。也就是说，即使“AS-SELECTED”适配子获得了有用的亲和力和特异性，它在适应EAB平台时也可能失败，因为它缺乏支持EAB信号所需的结合诱导的构象变化。在这篇文章中，我们揭示了我们用来修饰适配子的光谱指导方法，以便它们支持必要的结合诱导的构象变化。具体地说，使用新报道的适体，我们展示了EAB传感器的系统设计，实现了临床和生理上相关的特异性、检测限和针对靶标甲氨蝶呤和色氨酸的动态范围。
Electrochemical aptamer-based (EAB) sensors utilize the binding-induced conformational change of an electrode-attached, redox-reporter-modified aptamer to transduce target recognition into an easily measurable electrochemical output. Because this signal transduction mechanism is single-step and rapidly reversible, EAB sensors support high-frequency, real-time molecular measurements, and because it recapitulates the reagentless, conformation-linked signaling seen in vivo among naturally occurring receptors, EAB sensors are selective enough to work in the complex, time-varying environments found in the living body. The fabrication of EAB sensors, however, requires that their target-recognizing aptamer be modified such that (1) it undergoes the necessary binding-induced conformational change and (2) that the thermodynamics of this “conformational switch” are tuned to ensure that they reflect an acceptable trade-off between affinity and signal gain. That is, even if an “as-selected” aptamer achieves useful affinity and specificity, it may fail when adapted to the EAB platform because it lacks the binding-induced conformational change required to support EAB signaling. In this paper we reveal the spectroscopy-guided approaches we use to modify aptamers such that they support the necessary binding-induced conformational change. Specifically, using newly reported aptamers, we demonstrate the systematic design of EAB sensors achieving clinically and physiologically relevant specificity, limits of detection, and dynamic range against the targets methotrexate and tryptophan.
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影响因子：
7.4
作者：
Arroyo-Curras, Netzahualcoyotl;Scida, Karen;Plaxco, Kevin W.
通讯作者：
Plaxco, Kevin W.
影响因子：
15
作者：
Watkins, Herschel M.;Vallee-Belisle, Alexis;Ricci, Francesco;Makarov, Dmitrii E.;Plaxco, Kevin W.
通讯作者：
Plaxco, Kevin W.
影响因子：
9.5
作者：
Nakatsuka N;Abendroth JM;Yang KA;Andrews AM
通讯作者：
Andrews AM
DOI：
10.1126/science.aao6750
发表时间：
2018-10-19
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Nakatsuka N;Yang KA;Abendroth JM;Cheung KM;Xu X;Yang H;Zhao C;Zhu B;Rim YS;Yang Y;Weiss PS;Stojanović MN;Andrews AM
通讯作者：
Andrews AM
影响因子：
15
作者：
Xiao, Y;Piorek, BD;Heeger, AJ
通讯作者：
Heeger, AJ