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Sensors 2017, 17(9), 2044; https://doi.org/10.3390/s17092044

Aptamer-Based Single-Step Assay by the Fluorescence Enhancement on Electroless Plated Nano Au Substrate

1
Center for BioMicrosystems, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Sungbuk-gu, Seoul 136-791, Korea
2
School of Micro Nano System Engineering, Korea University of Science and Technology, Daejeon 305-333, Korea
3
School of Chemical Engineering, Birla Institute of Technology and Science, Pilani, K. K. Birla Goa Campus, Zuari Nagar, Goa 403-726, India
*
Author to whom correspondence should be addressed.
Received: 26 July 2017 / Revised: 25 August 2017 / Accepted: 2 September 2017 / Published: 7 September 2017
(This article belongs to the Section Biosensors)
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Abstract

A new single-step aptamer-based surface-enhanced fluorescent optical sensor is built, by combining an aptamer–target interaction for target recognition and a fluorophore interaction for signal enhancement. The developed aptasensor is simple, sensitive, specific and stable for the detection of thrombin. A new nanometallic Au structure in the range of 100 nm was constructed through effective electroless plating method on a Cu thin film. Cu+ ions act as sacrificial seeds for the reduction of Au2+/3+ ions to form Au nanolawns. In order to utilize the structure for a fluorescence-based sensor, aptamer conjugated with Cy3 was immobilized on the nanogold substrate through electrostatic attraction. The Au substrate was coated with chitosan (molecular weight 1000 Da). Thrombin binding aptamer (TBA) was applied as a model system demonstrating the aptamer-based fluorescence assay on nanogold substrates. Thrice-enhanced fluorescence emission was achieved with Cy3-conjugated TBA stably immobilized on the chitosan-coated Au substrate. The intensity change was proportional to the concentration of thrombin from 10 μM to 10 pM, whereas the intensity change was ignorable for other proteins such as human serum albumin (HSA). Aptamer-based assay benefited from simple immobilization of receptors and Au nanostructure contributed in building an effective surface enhancing/positively charged substrate was proved. Such an aptasensor holding high utilities for point-of-care devices by incorporating simplicity, sensitivity and selectivity in detection, low-cost for test, small sample volumes has been developed. View Full-Text
Keywords: aptamer; electroless deposition; gold nanostructure; thrombin; human serum albumin; fluorescence; optical biosensor aptamer; electroless deposition; gold nanostructure; thrombin; human serum albumin; fluorescence; optical biosensor
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Nambi Krishnan, J.; Park, S.-H.; Kim, S.K. Aptamer-Based Single-Step Assay by the Fluorescence Enhancement on Electroless Plated Nano Au Substrate. Sensors 2017, 17, 2044.

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