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Label-Free Detection of Peptide Nucleic Acid-DNA Hybridization Using Localized Surface Plasmon Resonance Based Optical Biosensor

机译:基于局部表面等离振子共振的光学生物传感器的肽核酸-DNA杂交的无标记检测。

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摘要

The development of label-free optical biosensors for DNA and other biomolecules has the potential to impact life sciences as well as screening in medical and environmental applications. In this report, we developed a localized surface plasmon resonance (LSPR) based label-free optical biosensor based on a gold-capped nanoparticle layer substrate immobilized with peptide nucleic acids (PNAs). PNA probe was designed to recognize the target DNA related to tumor necrosis factor. The nanoparticle layer was formed on a gold-deposited glass substrate by the surface modified silica nanoparticles using silane-coupling reagent. The optical properties of gold-capped nanoparticle layer substrate were characterized through monitoring the changes in the absorbance strength, as the thickness of the biomolecular layer increased with hybridization. The detection of PNA-DNA hybridization with target oligonucleotides and PCR-amplified real samples were performed with a limit of detection value of 0.677 pM target DNA. Selective discrimination against a singlebase mismatch was also achieved. Our LSPR-based biosensor with the gold-capped nanoparticle layer substrate is applicable to the design of biosensors for monitoring of the interaction of other biomolecules, such as proteins, whole cells, or receptors with a massively parallel detection capability in a highly miniaturized package.
机译:用于DNA和其他生物分子的无标签光学生物传感器的开发具有影响生命科学以及在医学和环境应用中进行筛选的潜力。在此报告中,我们基于固定有肽核酸(PNA)的金盖纳米颗粒层基质,开发了基于局部表面等离子体共振(LSPR)的无标记光学生物传感器。 PNA探针旨在识别与肿瘤坏死因子相关的目标DNA。使用硅烷偶联剂通过表面改性的二氧化硅纳米粒子在沉积金的玻璃基板上形成纳米粒子层。随着生物分子层厚度的增加,通过监测吸光度强度的变化来表征金包覆的纳米颗粒层基板的光学性能。用靶寡核苷酸和PCR扩增的真实样品进行PNA-DNA杂交的检测,检测限为0.677 pM靶DNA。还实现了针对单碱基不匹配的选择性鉴别。我们的基于LSPR的生物传感器具有镀金的纳米颗粒层基底,适用于生物传感器的设计,该传感器可在高度小型化的包装中监视具有大规模并行检测能力的其他生物分子(例如蛋白质,全细胞或受体)的相互作用。

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