A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes
A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes
复制标题
DOI：
10.1021/jacs.9b06229
复制
发表时间：
2019-07-24
影响因子：
15
通讯作者：
Easley, Christopher J.
中科院分区：
文献类型：
--
作者：
Somasundaram, Subramaniam;Easley, Christopher J.
为了使检测在床旁临床分析中最有效，它需要经济、简单、可推广，并且没有繁琐的工作流程。虽然基于电化学的DNA传感器降低了仪器成本并消除了复杂的程序，但仍然需要解决探针成本和通用性，因为需要具有多个缀合的许多探针来量化广泛的生物标志物。在这项工作中，我们已经开辟了一条路线，以规避复杂的多共轭计划，直接在电极表面上使用酶催化的探针建设。有了这个，我们已经创建了一个通用的DNA纳米结构探针，并验证其有效性，通过定量蛋白质（链霉亲和素，抗地高辛，抗他克莫司）和小分子（生物素，地高辛，他克莫司）使用相同的平台。他克莫司是一种广泛用于器官移植患者的免疫抑制药物，首次采用电化学直接定量，测定范围与治疗指数范围相匹配。最后，在最低限度稀释的人血清的背景下证实了探针的稳定性和灵敏度。
For an assay to be most effective in point-of-care clinical analysis, it needs to be economical, simple, generalizable, and free from tedious workflows. While electrochemistry-based DNA sensors reduce instrumental costs and eliminate complicated procedures, there remains a need to address probe costs and generalizability, as numerous probes with multiple conjugations are needed to quantify a wide range of biomarkers. In this work, we have opened a route to circumvent complicated multi conjugation schemes using enzyme-catalyzed probe construction directly on the surface of the electrode. With this, we have created a versatile DNA nanostructure probe and validated its effectiveness by quantification of proteins (streptavidin, anti-digoxigenin, anti-tacrolimus) and small molecules (biotin, digoxigenin, tacrolimus) using the same platform. Tacrolimus, a widely prescribed immunosuppressant drug for organ transplant patients, was directly quantified with electrochemistry for the first time, with the assay range matching the therapeutic index range. Finally, the stability and sensitivity of the probe was confirmed in a background of minimally diluted human serum.