Grafting of a peptide probe for Prostate-Specific Antigen detection using diazonium electroreduction and click chemistry

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 t...

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Bibliographic Details
Published in:	Biosensors & bioelectronics Vol. 81; pp. 131 - 137
Main Authors:	Strzemińska, I., Sainte Rose Fanchine, S., Anquetin, G., Reisberg, S., Noël, V., Pham, M.C., Piro, B.
Format:	Journal Article
Language:	English
Published:	England Elsevier B.V 15-07-2016
Subjects:	Amino Acid Sequence Aniline Aniline Compounds - chemistry Antigens Azo Compounds - chemistry Biosensing Techniques - methods Biosensors Click Chemistry Diazonium Compounds - chemistry Diazonium salts Electrochemical peptide sensor Electrochemical Techniques - methods Electrodes Electroplating Electrowinning Humans Label-free detection Male Models, Molecular Molecular Probes - chemistry Naphthoquinones - chemistry Oxidation-Reduction Peptides Peptides - chemistry Prostate-specific Antigen (PSA) Prostate-Specific Antigen - analysis Synthesis (chemistry) Transducers Prostate-specific Antigen (PSA) Electrochemical peptide sensor Label-free detection Click chemistry Diazonium salts
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Summary:	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. Working principle for the detection of PSA using the peptide probe. [Display omitted] •Diazonium salts reduction and click chemistry for biofunctionalization.•Reagentless and label-free electrochemical peptide sensor for prostate specific antigen detection.•Peptide cleavage by prostate specific antigen.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0956-5663 1873-4235
DOI:	10.1016/j.bios.2016.02.060