Department of Micro- and Nanotechnology, Technical University of Denmark, Lyngby, DK-2800, Denmark.
Nanotechnology. 2013 Nov 8;24(44):444011. doi: 10.1088/0957-4484/24/44/444011. Epub 2013 Oct 10.
In this work the use of a high-throughput nanomechanical detection system based on a DVD-ROM optical drive and cantilever sensors is presented for the detection of urokinase plasminogen activator receptor inflammatory biomarker (uPAR). Several large scale studies have linked elevated levels of soluble uPAR (suPAR) to infectious diseases, such as HIV, and certain types of cancer. Using hundreds of cantilevers and a DVD-based platform, cantilever deflection response from antibody-antigen recognition is investigated as a function of suPAR concentration. The goal is to provide a cheap and portable detection platform which can carry valuable prognostic information. In order to optimize the cantilever response the antibody immobilization and unspecific binding are initially characterized using quartz crystal microbalance technology. Also, the choice of antibody is explored in order to generate the largest surface stress on the cantilevers, thus increasing the signal. Using optimized experimental conditions the lowest detectable suPAR concentration is currently around 5 nM. The results reveal promising research strategies for the implementation of specific biochemical assays in a portable and high-throughput microsensor-based detection platform.
在这项工作中,我们使用了一种基于 DVD-ROM 光驱和悬臂传感器的高通量纳米机械检测系统,用于检测尿激酶型纤溶酶原激活物受体炎症生物标志物 (uPAR)。多项大规模研究表明,可溶性 uPAR (suPAR) 水平升高与 HIV 等传染病和某些类型的癌症有关。我们使用数百个悬臂和基于 DVD 的平台,研究了抗体-抗原识别的悬臂挠度响应随 suPAR 浓度的变化。我们的目标是提供一种廉价且便携的检测平台,能够提供有价值的预后信息。为了优化悬臂的响应,我们最初使用石英晶体微天平技术对抗体固定和非特异性结合进行了表征。此外,我们还探索了抗体的选择,以在悬臂上产生最大的表面应力,从而提高信号。在优化实验条件下,目前最低可检测的 suPAR 浓度约为 5 nM。这些结果为在便携式高通量基于微传感器的检测平台上实现特定生化分析提供了有前景的研究策略。