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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Ocaña, Cristina Del Valle, Manel |
| Description | Country affiliation: Spain Author Affiliation: Ocaña C ( Sensors and Biosensors Group, Department of Chemistry, Universitat Autonoma de Barcelona, 08193 Bellaterra, Spain.); del Valle M ( Sensors and Biosensors Group, Department of Chemistry, Universitat Autonoma de Barcelona, 08193 Bellaterra, Spain. Electronic address: manel.delvalle@uab.cat.) |
| Abstract | In this work, we report a highly specific amplification strategy demonstrated for the ultrasensitive biosensing of thrombin with the use of gold-streptavidin nanoparticles (strep-AuNPs) and silver reduction enhancement. The biotinylated aptamer of thrombin was immobilized onto an avidin-graphite epoxy composite (AvGEC) electrode surface by affinity interaction between biotin and avidin; electrochemical impedance measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The change in interfacial charge transfer resistance (Rct) experimented by the redox marker, was recorded to confirm aptamer complex formation with target protein, thrombin (Thr), in a label-free first stage. A biotinylated second thrombin aptamer, with complementary recognition properties was then used in a sandwich approach. The addition of strep-AuNPs and silver enhancement treatment led to a further increment of Rct thus obtaining significant signal amplification. The AptThrBio1-Thr-AptThrBio2 sandwich formation was inspected by confocal microcopy after incubation with streptavidin quantum dots. In order to visualize the presence of gold nanoparticles, the same silver enhancement treatment was applied to electrodes already modified with the nanoparticle-sandwich conjugate, allowing direct observation by scanning electron microscopy (SEM). Results showed high sensitivity and selectivity for thrombin detection, with an improvement from ca. 4.7 pM in a simple assay to 0.3 pM in the amplified reported scheme. |
| ISSN | 09565663 |
| Volume Number | 54 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2014-04-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Aptamers, Nucleotide Chemistry Biosensing Techniques Instrumentation Gold Nanoparticles Streptavidin Thrombin Analysis Electric Impedance Equipment Design Limit Of Detection Quantum Dots Evaluation Studies Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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