Development of Metal-Enhanced Fluorescence-Based Aptasensor for Thrombin Detection Using Silver Dendritic Nanostructures
- 45 Downloads
Metal-enhanced fluorescence (MEF) phenomenon has shown a promising potential in the field of fluorescence-based biological sensing. In this study, we optimized the electroless metal deposition method to fabricate silver dendritic nanostructures as effective MEF active substrates. Then, an aptasensor was developed for thrombin detection using the established surfaces. For this purpose, thiolated 29-mer thrombin-binding aptamers (TBA29 (12T) SH) as capturing aptamer were immobilized on the surface of silver dendritic nanostructures, then thrombin was sandwiched between the capturing aptamer and Cy5-labeled 15-mer thrombin aptamer (TBA15-Cy5). Quantitative analysis was performed through fluorescence signal measurement. The established aptasensor presented satisfactory sensitivity and selectivity and exhibited a limit of detection (LOD) as low as 32 pM. This aptasensor was also able to detect thrombin in the human serum at picomolar levels. Furthermore, the ease and relatively low-cost of fabrication of this platform introduce it as a tool with great potential for the clinical diagnosis of diseases and also for improving sensitivity of a variety of technologies which exploit fluorescent dyes for analyte detection, at ultra-trace levels, in complex matrices.
KeywordsSilver dendritic nanostructure Metal-enhanced fluorescence (MEF) Thrombin Aptamer
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
The experimental protocol was approved by the research ethics committee of Tarbiat Modares University. The participant provided written informed consent.
- 1.Hall EAH (1991) Biosensors. Prentice-Hall, Englewood CliffsGoogle Scholar
- 8.Goldys EM (2009) Fluorescence applications in biotechnology and life sciences. Wiley, HobokenGoogle Scholar
- 10.Tansakul C, Lilie E, Walter ED, Rivera F III, Wolcott A, Zhang JZ, Millhauser GL, Braslau R (2010) Distance-dependent fluorescence quenching and binding of CdSe quantum dots by functionalized nitroxide radicals. J Phys Chem C Nanomater Interfaces 114:7793–7805CrossRefPubMedPubMedCentralGoogle Scholar
- 25.Le Ru EC, Etchegoin PG (2005) Surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF) in the context of modified spontaneous emission. arXiv:physics/0509154 [https://www.physics.chem-ph]
- 53.He L, Lin M, Li H, Kim N (2010) Surface-enhanced Raman spectroscopy coupled with dendritic silver nanosubstrate for detection of restricted antibiotics. J Raman Spectrosc 41:739–744Google Scholar
- 56.Little TA (2015) Method validation essentials, limit of blank, limit of detection, and limit of quantitation. BioPharm Int 28:48–51Google Scholar