Mei Zhong, Wang Yanyan, Tang Liang
Department of Biomedical Engineering, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX, 78249, USA.
Methods Mol Biol. 2017;1571:129-141. doi: 10.1007/978-1-4939-6848-0_9.
Gold nanorod (GNR) based label-free sensing has been attractive due to its unique property of localized surface plasmon resonance (LSPR). Compared to bulk gold, the SPR of GNRs is more sensitive to the refractive index change caused by biological binding in the close proximity. Numerous studies have reported biological detection in solution based GNR probes. However, the biosensing has the intrinsic problems of fluctuating readings and short storage time due to nanoparticle aggregation. In contrast, a chip-based nanorod biosensor is a more robust and reliable platform. We have developed a nanoplasmonic biosensor in a chip format by immobilizing functionalized GNRs on a (3-mercaptopropyl)trimethoxysilane modified glass substrate. The covalent Au-S bond ensures a strong GNR deposition on the substrate. This biochip exhibits a high sensitivity and stability when exposed to physiological buffer with high ionic strength. Another advantage of GNR as optical transducer is its LSPR peak dependence on the aspect ratio, which provides an ideal multiplexed detection mechanism. GNRs of different sizes that exhibit distinct SPR peaks are combined and deposited on designated spots of a glass substrate. The spectral shift of the respective peaks upon the biological binding are monitored for simultaneous detection of specific analytes. Coupled with a microplate reader, this spatially resolved GNR array biochip results in a high-throughput assay of samples as well as multiplexed detection in each sample. Since most biological molecules such as antibodies and DNA can be linked to GNR using previously reported surface chemistry protocol, the label-free nanosensor demonstrated here is an effective tool for protein/DNA array analysis, especially for detection of disease biomarkers.
基于金纳米棒(GNR)的无标记传感因其独特的局域表面等离子体共振(LSPR)特性而备受关注。与块状金相比,GNR的表面等离子体共振对近距离生物结合引起的折射率变化更为敏感。许多研究报道了基于溶液的GNR探针用于生物检测。然而,由于纳米颗粒聚集,这种生物传感存在读数波动和储存时间短的固有问题。相比之下,基于芯片的纳米棒生物传感器是一个更稳健、可靠的平台。我们通过将功能化的GNR固定在(3-巯基丙基)三甲氧基硅烷修饰的玻璃基板上,开发了一种芯片形式的纳米等离子体生物传感器。共价Au-S键确保了GNR在基板上的牢固沉积。当暴露于高离子强度的生理缓冲液时,这种生物芯片表现出高灵敏度和稳定性。GNR作为光学换能器的另一个优点是其LSPR峰依赖于纵横比,这提供了一种理想的多重检测机制。将具有不同SPR峰的不同尺寸的GNR组合并沉积在玻璃基板的指定位置。监测生物结合后各个峰的光谱位移,以同时检测特定分析物。结合酶标仪,这种空间分辨的GNR阵列生物芯片可实现样品的高通量检测以及每个样品中的多重检测。由于大多数生物分子如抗体和DNA可以使用先前报道的表面化学方法与GNR连接,这里展示的无标记纳米传感器是蛋白质/DNA阵列分析的有效工具,特别是用于疾病生物标志物的检测。