Liang Ru-Qiang, Tan Cui-Yan, Ruan Kang-Cheng
Key Laboratory of Proteomics, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China.
J Immunol Methods. 2004 Feb 15;285(2):157-63. doi: 10.1016/j.jim.2003.11.008.
This work presents a method for analyzing protein microarrays using a colorimetric nanogold probe coupled with silver enhancement (gold-silver detection). In this method, the gold nanoparticles were introduced to the microarray by the specific binding of the gold-conjugated antibodies or streptavidins and then coupled with silver enhancement to produce black image of microarray spots, which can be easily detected with a commercial CCD camera. The method showed high detection sensitivity (1 pg of IgG immobilized on slides or 2.75 ng/ml IgG in solution) and a good linear correlation between the signal intensity and the logarithm of the sample concentration. The examination of this method in analyzing a demonstrational ToRCH antigen microarray developed in our lab showed an identical result as in the fluorescent method. These results suggest the colorimetric gold-silver detection method has potential applications in proteomics research and clinical diagnosis.
这项工作提出了一种使用比色纳米金探针结合银增强技术（金银检测法）分析蛋白质微阵列的方法。在该方法中，通过金偶联抗体或链霉亲和素的特异性结合将金纳米颗粒引入微阵列，然后与银增强技术相结合，产生微阵列斑点的黑色图像，该图像可通过商用CCD相机轻松检测。该方法显示出高检测灵敏度（固定在载玻片上的1 pg IgG或溶液中2.75 ng/ml IgG），并且信号强度与样品浓度的对数之间具有良好的线性相关性。在分析我们实验室开发的示范性ToRCH抗原微阵列时对该方法进行的检测显示，其结果与荧光法相同。这些结果表明，比色金银检测法在蛋白质组学研究和临床诊断中具有潜在应用。
