Previte Michael J R, Aslan Kadir, Geddes Chris D
Institute of Fluorescence, Laboratory for Advanced Medical Plasmonics, Medical Biotechnology Center, University of Maryland Biotechnology Institute, 725 West Lombard Street, Baltimore, Maryland 21201, USA.
Anal Chem. 2007 Sep 15;79(18):7042-52. doi: 10.1021/ac071042+. Epub 2007 Aug 16.
We have combined the principles of microwave circuitry and antenna design and our recent work in microwave-triggered metal-enhanced chemiluminescence to now "trigger" chemically and enzyme-catalyzed chemiluminescent reactions with spatial and temporal control. With this technology platform, we achieve spatial and temporal control of enzyme and chemically catalyzed chemiluminescence reactions to achieve more than 500-fold increases in "on-demand" photon flux from chemically catalyzed chemiluminescent reactions. We also report a 6-fold increase in photon flux from HRP-catalyzed assays on disposable coverslips functionalized with HRP and placed proximal to the substrates modified with thin-film aluminum triangle disjointed "bow-tie" structures. In addition, we demonstrate the applicability of this technology to develop multiplexed or high-throughput chemiluminescent assays. We also demonstrate the clinical and biological relevance of this technology platform by affixing aluminum structures in proximity to HRP protein immobilized on nitrocellulose to improve the sensitivity for this model Western blot scheme by 50-fold. We believe analytical applications that rely on enzyme-catalyzed chemiluminescence, such as immunoassays, may greatly benefit from this new platform technology.
我们将微波电路原理与天线设计以及我们近期在微波触发的金属增强化学发光方面的工作相结合,从而能够在空间和时间上对化学及酶催化的化学发光反应进行“触发”。借助这个技术平台,我们实现了对酶促和化学催化化学发光反应的时空控制,使化学催化化学发光反应的“按需”光子通量增加了500多倍。我们还报告称,在用HRP功能化并放置在经薄膜铝三角形不连续“领结”结构修饰的底物附近的一次性盖玻片上进行HRP催化检测时,光子通量增加了6倍。此外,我们展示了这项技术在开发多重或高通量化学发光检测方面的适用性。我们还通过将铝结构靠近固定在硝酸纤维素上的HRP蛋白来证明该技术平台的临床和生物学相关性,使这种模型蛋白质印迹方案的灵敏度提高了50倍。我们相信,依赖酶催化化学发光的分析应用,如免疫分析,可能会从这项新的平台技术中大大受益。