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方便地将 DNA 模板量子点掺入其中,用于灵敏电化学检测口腔癌生物标志物白细胞介素-8。

Facile incorporation of DNA-templated quantum dots for sensitive electrochemical detection of the oral cancer biomarker interleukin-8.

机构信息

West China School of Public Health and West China Fourth Hospital, Sichuan University, Chengdu, 610041, Sichuan, China.

School of Life Sciences, Shanghai University, Shanghai, 200444, China.

出版信息

Anal Bioanal Chem. 2020 Apr;412(11):2599-2606. doi: 10.1007/s00216-020-02487-x. Epub 2020 Feb 13.

Abstract

Recent studies reveal a great value of interleukin-8 (IL-8), a pro-inflammatory cytokine, as a potent biomarker for early diagnosis of oral cancer. Herein, a new electrochemical method is proposed to detect IL-8 by facilely incorporating DNA-templated quantum dots (QDs). In principle, target IL-8 is first treated with the reducing agent tris(2-carboxyethyl)phosphine (TCEP) to yield active thiols and then captured by antibody-functionalized magnetic beads (MBs). Thereafter, via the Michael addition reaction between the active thiol and maleimide group, a maleimide-modified DNA probe is linked to the surface of MBs, which can initiate a process of rolling circle amplification. In this way, long-range DNA strands are generated on the MB surface, subsequently recruiting DNA-templated CdTe/CdS QDs (DNA-QDs) to act as electrochemical reporters. By tracing the responses of DNA-QDs, the method allows IL-8 detection in a linear range from 5 to 5000 fg/mL with a detection limit of 3.36 fg/mL. The selectivity, reproducibility, and applicability in complex serum samples are also demonstrated to be favorable, indicating that the method may have a great potential in the future. More importantly, the use of TCEP treatment in the method not only provides a facile way to incorporate DNA-QDs, avoiding the complicated and time-consuming preparation process of antibody-DNA conjugates or functional nanomaterials; but also makes the method capable of being extended to detect other protein biomarkers in view of widespread presence of disulfides, which may hold a broad potential to facilitate efficient biosensing designs.

摘要

最近的研究揭示了白细胞介素-8(IL-8)作为一种强有力的生物标志物在口腔癌早期诊断中的巨大价值。在这里,提出了一种新的电化学方法,通过简便地将 DNA 模板量子点(QDs)掺入来检测 IL-8。从原则上讲,首先用还原剂三(2-羧乙基)膦(TCEP)处理靶标 IL-8,以生成活性硫醇,然后用抗体功能化的磁性珠(MBs)捕获。此后,通过活性硫醇和马来酰亚胺之间的迈克尔加成反应,将马来酰亚胺修饰的 DNA 探针连接到 MBs 的表面,这可以引发滚环扩增过程。通过在 MB 表面上生成长程 DNA 链,随后招募 DNA 模板化 CdTe/CdS QDs(DNA-QDs)作为电化学报告器。通过跟踪 DNA-QDs 的响应,该方法允许在 5 至 5000 fg/mL 的线性范围内检测 IL-8,检测限为 3.36 fg/mL。还证明了该方法在复杂血清样品中的选择性、重现性和适用性良好,表明该方法在未来可能具有很大的潜力。更重要的是,该方法中 TCEP 处理的使用不仅提供了一种简便的方法来掺入 DNA-QDs,避免了抗体-DNA 缀合物或功能纳米材料的复杂和耗时的制备过程;而且由于二硫键的广泛存在,该方法能够扩展到检测其他蛋白质生物标志物,这可能为促进高效生物传感设计提供广泛的潜力。

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