Metal-enhanced fluorescent probes based on silver nanoparticles and its application in IgE detection

Anal Bioanal Chem. 2012 Jan;402(3):1057-63. doi: 10.1007/s00216-011-5591-1. Epub 2011 Dec 8.

Abstract

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 μg/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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Aptamers, Nucleotide / chemistry
  • Carbocyanines / chemistry
  • Fluorescent Dyes / chemistry*
  • Humans
  • Immunoassay / methods*
  • Immunoglobulin E / analysis*
  • Immunoglobulin E / immunology
  • Limit of Detection
  • Nanoparticles / chemistry*
  • Nucleic Acid Hybridization
  • Silver / chemistry*

Substances

  • Antibodies, Immobilized
  • Aptamers, Nucleotide
  • Carbocyanines
  • Fluorescent Dyes
  • cyanine dye 5
  • Immunoglobulin E
  • Silver