Improving Pb2+ detection using DNAzyme-based fluorescence sensors by pairing fluorescence donors with gold nanoparticles.
For previously reported fluorescence Pb(2+) sensors, DNAzymes have lead to a significant increase in Pb(2+) detecting sensitivity and specificity. However, these sensors suffer from incomplete fluorescence quenching and require additional steps for annealing DNAzymes and substrates as well as for removing the uncoupled substrates. In this study, we successfully overcome these issues by immobilizing the substrate nucleic acids on gold nanoparticles through thiol linkages. The immobilization of the substrate molecules to the gold nanoparticles lead to almost-complete fluorescence quenching and fast Pb(2+) detection, without altering the Pb(2+) specificity of the DNAzymes. After optimizing the concentration of DNAzymes, reaction time and pH, we could detect Pb(2+) as low as 5 nM within 20 min without the preliminary and the post treatments. Considering the multi-color-fluorescence quenching capability of gold nanoparticles and the to-be-developed functional nucleic acids for other metal ions, this study could extend the application of DNAzymes to the detection of multiple heavy metal ions.
Kim
Joong Hyun
JH
BioNano Research Center, Korea Research Institute of Bioscience and Biotechnology, 111 Gwanhangno, Yuseong-gu, Daejeon 305-806, South Korea.
Han
Sang Ho
SH
Chung
Bong Hyun
BH
D015374:Biosensing Techniques / Q000295:instrumentation*; D021881:DNA, Catalytic / Q000737:chemistry*; D004579:Electron Transport; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D007854:Lead / Q000032:analysis* / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology / Q000295:instrumentation*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D021881:DNA, Catalytic; D007854:Lead; D006046:Gold