Small molecule electro-optical binding assay using nanopores
Small molecule electro-optical binding assay using nanopores
复制标题
DOI：
10.1038/s41467-019-09476-4
复制
发表时间：
2019-04-17
影响因子：
16.6
通讯作者：
Edel, Joshua B.
中科院分区：
文献类型：
--
作者：
Cai, Shenglin;Sze, Jasmine Y. Y.;Edel, Joshua B.
在单分子水平上识别短核酸和蛋白质是发展新的检测策略的主要推动力。纳米孔传感由于其无标记操作和单分子灵敏度而日益受到重视。然而，选择性地检测小分子仍然具有挑战性。在这里，我们建议将纳米孔的电传感模式与基于荧光的检测相结合。选择性是通过将分子信标、互补DNA或蛋白质嫁接到DNA分子载体上来实现的。我们证明，电和光通道之间的同步事件的部分可以用于执行单分子结合分析，而不需要直接标记分析物。这种策略可用于检测人体血清和尿液等复杂生物液体中的目标。未来对这项技术的优化可能会使定量蛋白质检测以及基因突变分析的新方法能够应用于下一代临床样本分析。
The identification of short nucleic acids and proteins at the single molecule level is a major driving force for the development of novel detection strategies. Nanopore sensing has been gaining in prominence due to its label-free operation and single molecule sensitivity. However, it remains challenging to detect small molecules selectively. Here we propose to combine the electrical sensing modality of a nanopore with fluorescence-based detection. Selectivity is achieved by grafting either molecular beacons, complementary DNA, or proteins to a DNA molecular carrier. We show that the fraction of synchronised events between the electrical and optical channels, can be used to perform single molecule binding assays without the need to directly label the analyte. Such a strategy can be used to detect targets in complex biological fluids such as human serum and urine. Future optimisation of this technology may enable novel assays for quantitative protein detection as well as gene mutation analysis with applications in next-generation clinical sample analysis.