Electrochemical DNA Sensors Based on MoS2-AuNPs for Polynucleotide Kinase Activity and Inhibition Assay
Electrochemical DNA Sensors Based on MoS2-AuNPs for Polynucleotide Kinase Activity and Inhibition Assay
复制标题
基于 MoS2-AuNPs 的电化学 DNA 传感器，用于多核苷酸激酶活性和抑制测定
DOI：
10.1021/acsami.0c13385
复制
发表时间：
2020-10-14
影响因子：
9.5
通讯作者：
Xia，Fan
中科院分区：
文献类型：
--
作者：
Lin，Meihua;Wan，Hao;Xia，Fan
T4多核苷酸激酶（PNK）活性的测定和PNK抑制剂的筛选对于疾病诊断和药物开发具有重要意义。已经开发了许多电化学策略用于灵敏地测量PNK活性和抑制。然而，它们通常遭受额外的标记和电化学读出的检测过程的多个步骤。在此，我们已经证明了电化学DNA（E-DNA）传感器的一步检测PNK与“信号”读出，而不需要额外的标签。在我们的设计中，高度可切换的双链DNA（dsDNA）探针被固定在金纳米颗粒修饰的二硫化钼纳米材料（MoS 2-AuNPs）上，该材料具有大的表面积和高电导率，用于提高PNK检测中的信号增益。该传感器的检测限为2.18 × 10-4U/mL。此外，该测定显示出高选择性，具有区分PNK与其他酶和蛋白质的能力，并且可以用于筛选抑制剂。所提出的传感器是易于操作的一步读出和强大的PNK检测在生物基质中，并显示出巨大的潜力，在床旁的护理在临床诊断和药物筛选。
The determination of T4 polynucleotide kinase (PNK) activity and the screening of PNK inhibitors are critical to disease diagnosis and drug discovery. Numerous electrochemical strategies have been developed for the sensitive measurement of PNK activity and inhibition. However, they often suffer from additional labels and multiple steps of the detection process for the electrochemical readout. Herein, we have demonstrated an electrochemical DNA (E-DNA) sensor for the one-step detection of PNK with “signal-on” readout with no need for additional labels. In our design, the highly switchable double-stranded DNA (dsDNA) probes are immobilized on the gold nanoparticle-decorated molybdenum disulfide nanomaterial (MoS2-AuNPs), which possesses large surface area and high conductivity for elevating the signal gain in the PNK detection. This signal-on E-DNA sensor integrated with MoS2-AuNPs exhibits a much higher sensitivity than that without MoS2-AuNPs, showing a detection limit of 2.18 × 10–4U/mL. Furthermore, this assay shows high selectivity, with the ability to discriminate PNK from other enzymes and proteins, and can be utilized to screen inhibitors. The proposed sensor is easy to operate with one-step readout and robust for PNK detection in the biological matrix and shows great potential for point-of-care in clinical diagnostics and drug screening.