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Ratiometric Electrochemical strategy for Sensitive Determination of Furin Activity Based on Dual Signal Amplification and Antifouling Nanosurface

Abstract

To develop sensitive and accurate method for Furin activity is still the bottleneck for understanding the role of Furin plays in cell-surface systems and even in Alzheimer’s disease. In this work, a ratiometric electrochemical biosensor was developed for sensitive and accurate determination of Furin activity in the cell based on dual signal amplification stemmed from peptide with multiplex response sites and the antifouling gold nano-bellflowers (GBFs). New peptide, HS-CMRVRR↓YKDFDFG (P3), was firstly designed to be selectively cleavage by Furin at site↓. More importantly, this peptide P3 constitutes three amino acid residues with -COOH group subsequently used to bind with response molecule of ferrocene, and can remarkably improve the determination sensitivity for about 2.3 fold. Meanwhile, GBFs stabilized by PEG was taken as a second element to magnify the signal of ferrocene group via large ratio surface area and good conductivity, as well as an antibiofouling nanosurface to reduce the biofouling of electrode surface in cells. This double amplification strategy can greatly enhance the sensitivity of Furin detection for 6.5-folds, which is favorable for detection of low amounts Furin. In addition, 5’-MB-GGCGCGA(T)13-SH-3’ was co-assembled as an inner reference to provide a built-in element to correct the determination error resulting from complicate analysis environment. Finally, this sensitive and accurate Furin biosensor was successfully applied to detect Furin activity in Furin overexpressed U251 and MDA-MB-468 cells. As far as we know, it is the first report for electrochemical strategy to detect Furin activity in cell.

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Publication details

The article was received on 04 Aug 2017, accepted on 26 Sep 2017 and first published on 26 Sep 2017


Article type: Paper
DOI: 10.1039/C7AN01295K
Citation: Analyst, 2017, Accepted Manuscript
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    Ratiometric Electrochemical strategy for Sensitive Determination of Furin Activity Based on Dual Signal Amplification and Antifouling Nanosurface

    D. Yao , W. Zhao , L. Zhang and Y. Tian, Analyst, 2017, Accepted Manuscript , DOI: 10.1039/C7AN01295K

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