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利用压电纳米机械微悬臂梁的共振频率偏移进行前列腺特异性抗原(PSA)的免疫分析。

Immunoassay of prostate-specific antigen (PSA) using resonant frequency shift of piezoelectric nanomechanical microcantilever.

作者信息

Lee Jeong Hoon, Hwang Kyo Seon, Park Jaebum, Yoon Ki Hyun, Yoon Dae Sung, Kim Tae Song

机构信息

Microsystem Research Center, KIST, Seoul 136-791, Korea.

出版信息

Biosens Bioelectron. 2005 Apr 15;20(10):2157-62. doi: 10.1016/j.bios.2004.09.024.

Abstract

We designed and fabricated the nanomechanical Pb(Zr0.52Ti0.48)O3 (PZT) cantilever; we demonstrated a novel electrical measurement, under a controlled ambient temperature and humidity, for label-free detection of a prostate-specific antigen (PSA); and we achieved a detection sensitivity as low as 10 pg/ml. For the fabrication of our nanomechanical PZT cantilevers, we used composite layers of Ta/Pt/PZT/Pt/SiO2 on a SiN(x) supporting layer for electrical self-sensing without external oscillators. This method allows PSA proteins to be detected via a simple electrical measurement of the resonant frequency change generated by the molecular interaction of the antigen (Ag) and the antibody (Ab). The resonant frequency shifted due to the specific binding of the PSA Ag to its Ab which is immobilized via calixcrown self-assembled monolayers on an Au surface deposited on a nanomechanical PZT cantilever. We determined the resonant frequency shift as the value of -172 Hz and -273 Hz, when the concentration of PSA Ag was 1 ng/ml, with the cantilever dimensions of 100 microm x 300 microm and 50 microm x 150 microm, respectively. Theoretical and experimental analysis suggests that the minimum detectable sensitivity for a resonant frequency shift due to a PSA Ag-Ab interaction depends on the dimensions of the nanomechanical PZT cantilever. These results also demonstrate that the experimentally measured resonant frequency shift is larger than that calculated theoretically due to the compressive stress of the PSA Ag-Ab interaction.

摘要

我们设计并制造了纳米机械铅锆钛酸铅(Pb(Zr0.52Ti0.48)O3,PZT)悬臂梁;我们展示了一种在可控的环境温度和湿度下用于无标记检测前列腺特异性抗原(PSA)的新型电学测量方法;并且我们实现了低至10 pg/ml的检测灵敏度。对于我们的纳米机械PZT悬臂梁的制造,我们在氮化硅(SiN(x))支撑层上使用了Ta/Pt/PZT/Pt/SiO2复合层,以实现无需外部振荡器的电学自传感。这种方法允许通过简单地电学测量由抗原(Ag)和抗体(Ab)的分子相互作用产生的共振频率变化来检测PSA蛋白。由于PSA Ag与其通过杯芳烃自组装单层固定在沉积于纳米机械PZT悬臂梁上的金表面的Ab的特异性结合,共振频率发生了偏移。当PSA Ag的浓度为1 ng/ml时,对于尺寸分别为100微米×300微米和50微米×150微米的悬臂梁,我们测定共振频率偏移值分别为 -172 Hz和 -273 Hz。理论和实验分析表明,由于PSA Ag - Ab相互作用导致的共振频率偏移的最小可检测灵敏度取决于纳米机械PZT悬臂梁的尺寸。这些结果还表明,由于PSA Ag - Ab相互作用的压缩应力,实验测量的共振频率偏移大于理论计算值。

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