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A sensitive sandwich structures time-resolved fluorescence method for thrombin detection

Abstract

We designed a sensitive and specific time-resolved fluorescence assay for detection human thrombin. This design was based on the amplification of nanoparticles and the specificity of sandwich structure binding of two affinity aptamers. The DC@AuNPs was prepared by AuNPs conjugated to BSA-Eu3+-DTPA and SH-Apt29. In the presence of thrombin, the formation of quadruplexes-thrombin complex could lead to the formation of MNPs@Atp15-thrombin-DC@AuNPs sandwich structure. In the presence of enhancement solution, the Eu3+ was interacted with the enhancement solution to produce strong fluorescence. Under the optimized conditions, the linear range of the method was from 1 pM to 100 pM of thrombin, and the limit of detection was 0.78 pM. Furthermore, the method could specifically recognize thrombin from other analogous proteins. The method is successfully used to determine thrombin in human plasma. This method had the advantages on high sensitivity and selectivity. It showed great potential for medical diagnosis.

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Supplementary files

Publication details

The article was received on 19 May 2018, accepted on 01 Aug 2018 and first published on 08 Aug 2018


Article type: Paper
DOI: 10.1039/C8AY01135D
Citation: Anal. Methods, 2018, Accepted Manuscript
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    A sensitive sandwich structures time-resolved fluorescence method for thrombin detection

    Y. Zhang, T. Wu, T. Hou, Y. Zhang and S. Wang, Anal. Methods, 2018, Accepted Manuscript , DOI: 10.1039/C8AY01135D

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