Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University , Chongqing 400715, P.R. China.
Anal Chem. 2017 Aug 15;89(16):8266-8272. doi: 10.1021/acs.analchem.7b01272. Epub 2017 Jul 28.
In this work, an elegantly designed host-guest recognition-assisted electrochemical release was established and applied in a reusable electrochemical biosensor for the detection of microRNA-182-5p (miRNA-182-5p), a prostate cancer biomarker in prostate cancer, based on the DNA cross configuration-fueled target cycling and strand displacement reaction (SDR) amplification. With such a design, the single target miRNA input could be converted to large numbers of single-stranded DNA (S1-Trp and S2-Trp) output, which could be trapped by cucurbit[8]uril methyl viologen (CB-8-MV) based on the host-guest recognition, significantly enhancing the sensitivity for miRNA detection. Moreover, the nucleic acids products obtained from the process of cycling amplification could be utilized sufficiently, avoiding the waste and saving the experiment cost. Impressively, by resetting a settled voltage, the proposed biosensor could release S1-Trp and S2-Trp from the electrode surface, attributing that the guest ion methyl viologen (MV) was reduced to MV under this settled voltage and formed a more-stable CB-8-MV-MV complex. Once O was introduced in this system, MV could be oxidized to MV, generating the complex of CB-8-MV for capturing S1-Trp and S2-Trp again in only 5 min. As a result, the simple and fast regeneration of biosensor for target detection was realized on the base of electrochemical redox-driven assembly and release, overcoming the challenges of time-consuming, burdensome operations and expensive experimental cost in traditional reusable biosensors and updating the construction method for a reusable bisensor. Furthermore, the biosensor could be reused for more than 10 times with a regeneration rate of 93.20%-102.24%. After all, the conception of this work provides a novel thought for the construction of effective reusable biosensor to detect miRNA and other biomarkers and has great potential application in the area requiring the release of nucleic acids or proteins.
在这项工作中,建立了一种设计精巧的主体-客体识别辅助电化学释放,并将其应用于基于 DNA 交叉构型触发的目标循环和链置换反应(SDR)扩增的前列腺癌生物标志物 microRNA-182-5p(miRNA-182-5p)的可重复使用电化学生物传感器中。通过这种设计,单个目标 miRNA 的输入可以转换为大量的单链 DNA(S1-Trp 和 S2-Trp)输出,这些输出可以通过主体-客体识别被葫芦[8]脲甲基紫精(CB-8-MV)捕获,从而显著提高 miRNA 检测的灵敏度。此外,循环扩增过程中获得的核酸产物可以被充分利用,避免浪费并节省实验成本。令人印象深刻的是,通过重置设定电压,该生物传感器可以将 S1-Trp 和 S2-Trp 从电极表面释放出来,这是因为在设定电压下,客体离子甲基紫精(MV)被还原为 MV,并形成更稳定的 CB-8-MV-MV 配合物。一旦在该系统中引入 O,MV 可以被氧化为 MV,再次生成 CB-8-MV 复合物以捕获 S1-Trp 和 S2-Trp,仅需 5 分钟。因此,基于电化学氧化还原驱动的组装和释放,实现了生物传感器的简单快速再生,用于目标检测,克服了传统可重复使用生物传感器中耗时、繁琐的操作和昂贵的实验成本的挑战,并更新了可重复使用生物传感器的构建方法。此外,该生物传感器可以重复使用超过 10 次,再生率为 93.20%-102.24%。总之,这项工作的构想为构建用于检测 miRNA 和其他生物标志物的有效可重复使用生物传感器提供了新的思路,在需要释放核酸或蛋白质的领域具有巨大的潜在应用价值。