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使用基于局域表面等离子体共振的光学生物传感器对肽核酸- DNA杂交进行无标记检测。

Label-free detection of peptide nucleic acid-DNA hybridization using localized surface plasmon resonance based optical biosensor.

作者信息

Endo Tatsuro, Kerman Kagan, Nagatani Naoki, Takamura Yuzuru, Tamiya Eiichi

机构信息

Department of Biological Science and Biotechnology, School of Material Science, Japan Advanced Institute of Science and Technology, Nomi City, Ishikawa 923-1292, Japan.

出版信息

Anal Chem. 2005 Nov 1;77(21):6976-84. doi: 10.1021/ac0513459.

Abstract

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 single-base 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.

摘要

用于DNA和其他生物分子的无标记光学生物传感器的发展,有可能对生命科学以及医学和环境应用中的筛选产生影响。在本报告中,我们基于固定有肽核酸(PNA)的金帽纳米颗粒层基板,开发了一种基于局域表面等离子体共振(LSPR)的无标记光学生物传感器。PNA探针被设计用于识别与肿瘤坏死因子相关的目标DNA。纳米颗粒层通过使用硅烷偶联剂对表面进行修饰的二氧化硅纳米颗粒,在金沉积玻璃基板上形成。通过监测吸光度强度的变化来表征金帽纳米颗粒层基板的光学性质,因为随着杂交的进行生物分子层的厚度会增加。对与目标寡核苷酸的PNA-DNA杂交以及PCR扩增的实际样品进行了检测,目标DNA的检测限为0.677 pM。还实现了对单碱基错配的选择性区分。我们基于LSPR的带有金帽纳米颗粒层基板的生物传感器,适用于设计用于监测其他生物分子相互作用的生物传感器,例如蛋白质、全细胞或受体,具有在高度小型化封装中进行大规模平行检测的能力。

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