Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1163159 | Analytica Chimica Acta | 2016 | 7 Pages |
•A new signal inhibition mechanism was designed for electrochemical immunoassay.•Ferrocene serves as an ideal indicator to monitor the immunoassay process.•Horseradish peroxidase label induces insoluble deposition for signal inhibition.•The signal amplification of the nanoprobe greatly improves the sensitivity.•The positive detection potential range avoids the dissolved oxygen interference.
A nanoprobe-induced signal inhibition mechanism was designed for ultrasensitive electrochemical immunoassay at a chitosan-ferrocene (CS-Fc) based immunosensor. The nanoprobe was prepared by covalently loading signal antibody and high-content horseradish peroxidase (HRP) on the graphene oxide (GO) nanocarrier. The immunosensor was prepared through the stepwise assembly of gold nanoparticles (Au NPs) and capture antibody at a CS-Fc modified electrode. After sandwich immunoreaction, the GO-HRP nanoprobes were quantitatively captured onto the immunosensor surface and thus induced the production of a layer of insoluble film through the enzymatically catalytic reaction of the HRP labels. Both the dielectric immunocomplex formed on the immunosensor surface and the enzymatic precipitate with low electroconductivity led to the electrochemical signal decease of the Fc indicator, which was greatly amplified by the multi-enzyme signal amplification of the nanoprobe. Based on this amplified signal inhibition mechanism, a new ultrasensitive electrochemical immunoassay method was developed. Using carcinoembryonic antigen as a model analyte, this method showed a wide linear range over 5 orders of magnitude with a detection limit down to 0.54 pg/mL. Besides, the immunosensor showed good specificity, acceptable reproducibility and stability as well as satisfactory reliability for the serum sample analysis.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slide