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Highly Sensitive Optical Biosensor for Thrombin Based on Structure Switching Aptamer-Luminescent Silica Nanoparticles

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Abstract

We describe here the construction of a sensitive and selective optical sensor system for the detection of human α-thrombin. The surface functionalized luminescent [Ru(dpsphen)3]4− (dpsphen-4,7-diphenyl-1,10-phenanthroline disulfonate) ion doped silica nanoparticles (SiNPs) with a size ~70 nm have been prepared. The DABCYL (2-(4-dimethylaminophenyl)diazenyl-benzoic acid) quencher labeled thrombin binding aptamer is conjugated to the surface of SiNPs using BS3 (bis(sulfosuccinimidyl) suberate) as a cross-linker, resulting in the conformational change of aptamer to form G-quadruplex structure upon the addition of thrombin. The binding event is translated into a change in the luminescence intensity of Ru(II) complex via FRET mechanism, due to the close proximity of DABCYL quencher with SiNPs. The selective detection of thrombin using the SiNPs-aptamer system up to 4 nM is confirmed by comparing its sensitivity towards other proteins. This work demonstrates the application of simple aptamer-SiNPs conjugate as a highly sensitive system for the detection of thrombin and also it is highly sensitive towards thrombin in the presence of other proteins and complex medium such as BSA.

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Acknowledgments

This work was supported by Department of Science and Technology, New Delhi, India. We thank Prof. S. Krishnasamy and P. Manojkumar, School of Biotechnology, Madurai Kamaraj University for valuable discussions.

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  1. School of Chemistry, Madurai Kamaraj University, Madurai, 625 021, India

    Ethiraju Babu, Paulpandian Muthu Mareeswaran & Seenivasan Rajagopal

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  1. Ethiraju Babu
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Correspondence to Seenivasan Rajagopal.

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The detailed experimental scheme, SEM, EDX data and label free scheme with luminescence spectral studies in the presence of thrombin. (DOCX 1979 kb)

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Babu, E., Mareeswaran, P.M. & Rajagopal, S. Highly Sensitive Optical Biosensor for Thrombin Based on Structure Switching Aptamer-Luminescent Silica Nanoparticles. J Fluoresc 23, 137–146 (2013). https://doi.org/10.1007/s10895-012-1127-0

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