Label-free detection of peptide nucleic acid-DNA hybridization using localized surface plasmon resonance based optical biosensor
Label-free detection of peptide nucleic acid-DNA hybridization using localized surface plasmon resonance based optical biosensor
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DOI：
10.1021/ac0513459
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发表时间：
2005-11-01
影响因子：
7.4
通讯作者：
Tamiya, E
中科院分区：
文献类型：
--
作者：
Endo, T;Kerman, K;Tamiya, E
DIVA和其他生物分子的无标记光学生物传感器的开发有可能影响生命科学以及医学和环境应用中的筛选。在这份报告中，我们开发了一个本地化的表面等离子体共振（LSPR）基于无标记的光学生物传感器的基础上的金帽纳米粒子层基板固定肽核酸（PNA）。设计PNA探针识别肿瘤坏死因子相关的靶DNA。通过使用硅烷偶联剂的表面改性的二氧化硅纳米颗粒在沉积金的玻璃基底上形成纳米颗粒层。通过监测吸光度强度的变化来表征金覆盖的纳米颗粒层基底的光学性质，因为生物分子层的厚度随着杂交而增加。用靶寡核苷酸和PCR扩增的真实的样品进行PNA-DNA杂交检测，检测限为0.677 pM靶DNA。也实现了对单碱基错配的选择性区分。我们的基于LSPR的生物传感器与金封端的纳米颗粒层基板适用于生物传感器的设计，用于监测其他生物分子的相互作用，如蛋白质，全细胞，或受体与大规模并行检测能力在一个高度小型化的包装。
The development of label-free optical biosensors for DIVA 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.