Journal
BIOSENSORS & BIOELECTRONICS
Volume 81, Issue -, Pages 131-137Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.02.060
Keywords
Electrochemical peptide sensor; Label-free detection; Prostate-specific Antigen (PSA); Click chemistry; Diazonium salts
Categories
Funding
- laboratory ITODYS
- scientific programm PHC GALILEE [32287TK]
- Sorbonne Paris Cite [ANR-11-IDEX-05-02]
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The main objective of this work was to validate a label-free electrochemical method of protein detection using peptides as capture probes. As a proof-of-concept, we used a 7 amino acids sequence (HSSKLQL) specific for Prostate Specific Antigen. We investigated various electrografting conditions of two anilines (2-[(4-aminophenyl)sulfanyl]-8-hydroxy-1,4-naphthoquinone and 4-azidoaniline) further converted in situ into their corresponding diazonium salts on glassy carbon electrodes. It was demonstrated that the best method to obtain a mixed layer is the simultaneous electroreduction of the two diazonium salts. 4-azidoaniline was used to covalently immobilize the ethynyl-functionalized peptide probe by click coupling, and the hydroxynaphthoquinone derivative plays the role of electrochemical transducer of the peptide-protein recognition. The proteolytic activity of PSA towards a small peptide substrate carrying streptavidin at its distal end was also investigated to design an original sensing architecture leading to a reagentless, label free, and signal-on PSA sensor. Without optimization, the limit of quantification can be estimated in the nM to pM range. (C) 2016 Elsevier B.V. All rights reserved.
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