An aptamer-based assay for thrombin via structure switch based on gold nanoparticles and magnetic nanoparticles.
An aptamer-based assay for thrombin with high specificity and sensitivity was presented. In the protocol, the aptamer for thrombin was immobilized on magnetic nanoparticle, and its complementary oligonucleotide was labeled with gold nanoparticles, then the aptamer was hybridized with the complementary oligonucleotide to form the duplex structure as a probe, this probe could be used for the specific recognition for thrombin. In the presence of thrombin, the aptamer prefer to form the G-quarter structure with thrombin, resulting in the dissociation of the duplex of the probe and the release of the gold labeled oligonucleotide. Upon this, we were able to detect thrombin through the detection of the electrochemical signal of gold nanoparticles. The strategy combines with the high specificity of aptamer and the excellent characteristics of nanoparticles. This assay is simple, rapid, sensitive and highly specific, it does not require labeling of thrombin, and it could be applied to detect thrombin in complex real sample. The method shows great potential in other protein analysis and in disease diagnosis.
Zheng
Jing
J
Department of Chemistry, East China Normal University, Shanghai 200062, China. kkzhengjing707@163.com
Cheng
Gui-Fang
GF
He
Pin-Gang
PG
Fang
Yu-Zhi
YZ
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D008280:Magnetics; D008855:Microscopy, Electron, Scanning; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013696:Temperature; D013917:Thrombin / Q000032:analysis* / Q000378:metabolism
D052157:Aptamers, Nucleotide; D006046:Gold; D013917:Thrombin