期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 402, 期 3, 页码 1057-1063出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-011-5591-1
关键词
Metal-enhanced fluorescence; Silver nanoparticles; Aptamer; IgE
资金
- National Basic Research Program of China (973 Program) [2011CB911003, 2007CB936404]
- National Natural Foundation of China [20975050, 21175066, 81173023]
- National Science Funds for Creative Research Groups [20821063]
- University of Guam
In this paper, a novel metal plasmon coupled with an aptamer-nucleotide hybridized probe was fabricated and applied for protein detection. The specific aptamer and single-strand oligonucleotide were chemically bound to silver nanoparticles (AgNPs), and Cy5-labeled, complementary single-strand oligonucleotides were hybridized with the particle-bound oligonucleotides. The hybridized DNA duplexes were regarded as rigid rods that separated the fluorophore Cy5 and the surface of AgNPs to reduce the competitive quenching. Using a model system comprising human immunoglobulin E (IgE) as the analyte and goat antihuman IgE as immobilized capture antibody on glass slides, we demonstrate that the detection performance of the synthetic probe was superior to the aptamer-based fluorescent probes. The results showed a good linear correlation for human IgE in the range from 10 ng/ml to 6.25 mu g/ml. The detection limit obtained was 1 ng/ml, which was 50 times lower than that using Cy5 oligonucleotide/aptamer hybrid duplex (Probe2) due to the metal-enhanced fluorescence effect. This new strategy opens the possibility for the preparation of high-sensitivity detection probes based on metal nanoparticles.
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