title: Grafting of a peptide probe for Prostate-Specific Antigen detection using diazonium electroreduction and click chemistry.
abstract: 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.
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: I
authors.initials: I
authors.lastname: Strzemińska
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: S
authors.initials: S
authors.lastname: Sainte Rose Fanchine
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: G
authors.initials: G
authors.lastname: Anquetin
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: S
authors.initials: S
authors.lastname: Reisberg
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: V
authors.initials: V
authors.lastname: Noël
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France.
authors.forename: M C
authors.initials: MC
authors.lastname: Pham
authors.affiliation: Université Paris Diderot, Sorbonne Paris Cité, ITODYS, UMR 7086 CNRS, 15 rue J-A de Baïf, 75205 Paris Cedex 13, France. Electronic address: piro@univ-paris-diderot.fr.
authors.forename: B
authors.initials: B
authors.lastname: Piro
mesh_terms: D000595:Amino Acid Sequence; D000814:Aniline Compounds / Q000737:chemistry; D001391:Azo Compounds / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D057930:Click Chemistry; D003979:Diazonium Compounds / Q000737:chemistry; D055664:Electrochemical Techniques / Q000379:methods*; D004595:Electroplating; D006801:Humans; D008297:Male; D008958:Models, Molecular; D015335:Molecular Probes / Q000737:chemistry*; D009285:Naphthoquinones / Q000737:chemistry; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000032:analysis*
chemical_list: D000814:Aniline Compounds; D001391:Azo Compounds; D003979:Diazonium Compounds; D015335:Molecular Probes; D009285:Naphthoquinones; D010455:Peptides; C070827:4-azidoaniline; D017430:Prostate-Specific Antigen; C035342:1,4-naphthoquinone
keywords: Click chemistry; Diazonium salts; Electrochemical peptide sensor; Label-free detection; Prostate-specific Antigen (PSA)