生物素修饰纳米银探针的制备及在蛋白芯片可视化检测中的应用

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (11): 2184.

生物素修饰纳米银探针的制备及在蛋白芯片可视化检测中的应用

李慧1, 钟文英1, 许丹科2

1. 中国药科大学基础部分析化学教研室, 南京 210009;
2. 南京大学化学化工学院, 南京 210093

收稿日期:2010-03-12 出版日期:2010-11-10 发布日期:2010-11-10
通讯作者: 钟文英. E-mail: wyzhong@cpu.edu.cn；许丹科. E-mail: xudanke@nju.edu.cn
基金资助:
国家“九七三”计划项目(批准号: 2006CB910803)和蛋白质组学国家重点实验室开放课题资助.

Preparation of Biotinylated Silver Nanoparticles and Its Application of Visual Detection Method for Protein Chip

LI Hui1, ZHONG Wen-Ying1*, XU Dan-Ke2*

1. Department of Analytical Chemistry, Department of Basic Sciences, China Pharmaceutical University, Nanjing 210009, China;
2. School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China

Received:2010-03-12 Online:2010-11-10 Published:2010-11-10
Contact: ZHONG Wen-Ying. E-mail: wyzhong@cpu.edu.cn；XU Dan-Ke2. E-mail: xudanke@nju.edu.cn
Supported by:
国家“九七三”计划项目(批准号: 2006CB910803)和蛋白质组学国家重点实验室开放课题资助.

摘要/Abstract

摘要： 采用寡核苷酸为连接分子成功制备了生物素修饰的纳米银探针, 并建立了纳米银催化同种金属离子的特异性还原显色反应. 实验采用蛋白质芯片为分析工具, 以微量人IgG为蛋白分析模式研究了纳米银探针/氢醌/硝酸银体系的显色分析性能. 实验结果表明, 上述检测体系可对160 fg~100 pg含量范围内的微量蛋白显示可视化结果, 蛋白点的灰度值与其浓度具有良好的相关性, 最小蛋白检测量可达160 fg. 同时还开展了与商品化链亲和素纳米金/银增强试剂显色方法的对比实验, 结果表明, 本法制备的探针对蛋白的检出限降低了约40倍, 且具有存储稳定、反应快速等优点.

关键词: 生物素修饰纳米银, 氢醌/硝酸银, 微量蛋白检测, 蛋白芯片

Abstract: Biotinylated silver nanoparticles(Bio-AgNPs) were successfully prepared using oligonucelotide as coupling molecules. The resulted bio-AgNPs could be used for visual detection for protein arrays by a catalytical reaction with hydroquinone/AgNO3. To probe the feasibility of visual detection, human IgG was used as a model protein sample to be immobilized on the glass slides and bio-AgNPs were employed to couple with the protein via stripavaidin labeled anti-human IgG. The results show that the linear relationship of protein concentration is between 160 fg and 100 pg and the limit of detection is 160 fg(S/N=3). Compared with the method using SA-labeled gold nanoparticle or silver enhancement, the sensitivity of this method is increased about 40 fold. The presented method shows that its advantages including high sensitivity, stability and rapidity.

Key words: Biotinylated silver nanoparticle(Bio-AgNPs), Hydroquinone/AgNO3, Micro-protein detection, Protein chip

TrendMD:

李慧, 钟文英, 许丹科. 生物素修饰纳米银探针的制备及在蛋白芯片可视化检测中的应用. 高等学校化学学报, 2010, 31(11): 2184.

LI Hui, ZHONG Wen-Ying*, XU Dan-Ke*. Preparation of Biotinylated Silver Nanoparticles and Its Application of Visual Detection Method for Protein Chip. Chem. J. Chinese Universities, 2010, 31(11): 2184.

参考文献

[1]Deng X. Y., Gao D. J., Tian Y., Chen Y. H., Yu A. M., Zhang H. Q., Wang X. H., Chen Y.. Chem. Res. Chinese Universities[J], 2010, 26(1): 23—26 [2]Liu B. H., Tsao Z. J., Wang J. J., Yu F. Y.. Anal. Chem.[J], 2008, 80: 7029—7035 [3]Liang R. Q., Tan C. Y., Ruan K. C.. J. Immunol. Methods[J], 2004, 285: 157—163 [4]Duan L. L., Wang Y. F., Li S. S. C., Wan Z. X., Zhai J. X.. BMC Infectious Diseases[J], 2005, 5: 53— [5]Guo H. S., Zhang J. N., Yang D., Xiao P. F., He N. Y.. Colloid. Surf. B[J], 2005, 40: 195—198 [6]Gupta S., Huda S., Kilpatrick P. K., Velev O. D.. Anal. Chem.[J], 2007, 79: 3810—3820 [7]Taton T. A., MirkinC. A., Letsinger R. T.. Science[J], 2000, 289: 1757—1760 [8]Wei H., Chen C. G., Han B. Y., Wang E. K.. Anal. Chem.[J], 2008, 80: 7051—7055 [9]Thompson D. G., Enright A., Faulds K., Smith W. E., Graham D.. Anal. Chem.[J], 2008, 80: 2805—2810 [10]Lee J. S., Lyttonjean A. K. R., Hurst S. J., Mirkin C. A.. Nano Lett.[J], 2007, 7(7): 2112—2115 [11]Tokareava I., Hutter E.. J. Am. Chem. Soc.[J], 2004, 126: 15784—15789 [12]Sengupta A., Thai C. K., Sastry M. S. R., Matthaei J. F., Schwartz D. T., Davis E. J., Baneyx F.. Langmuir[J], 2008, 24: 2000—2008 [13]KaleleA S. A., Kundu A. A., Gosavi S. W., Deobagkar D. N., Deobagkar D. D., Kulkarni S. K.. Small[J], 2006, 2(3): 335—338 [14]Kim D., Daniel W. L., Mirkin C. A.. Anal. Chem.[J], 2009, 81: 9183—9187 [15]Munro C. H., Smith W. E., Garner M., Clarkson J., White P. C.. Langmuir[J], 1995, 11: 3712—372 [16]Li H., Sun Z. Y., Zhong W. Y., Xu D. K., Chen H. Y.. Anal. Chem.[J], 2010, 82: 5477—5483 [17]Joos T. O., Schrenk M., Hopfl P., Kroger K., Chowdhury U., Stoll D., Schorner D., Durr M., Herick K., Rupp S., Sohn K., Hammerle H.. Electrophoresis[J], 2000, 21: 2641—2650 [18]Holgate C. S., Jackson P., Cowen P. N., Bird C. C.. J. Histochem. Cytochem.[J], 1983, 31: 938—944 [19]Lackie P. M.. Histochem. Cell Biol.[J], 1996, 106: 9—17 [20]Xie F., Baker M . S., Goldys E. M.. J. Phys. Chem. B[J], 2006, 110: 23085—23091 [21]Chou L. Y. T., Fischer H. C., Perrault S. D., Chan W. C. W.. Anal. Chem.[J], 2009, 81: 4560—4565 [22]Li T., Guo L. P., Wang Z. X.. Biosens. Bioelectro.[J], 2008, 23: 1125—1130