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Chemosensors 2016, 4(4), 23; doi:10.3390/chemosensors4040023

Design of an Affibody-Based Recognition Strategy for Human Epidermal Growth Factor Receptor 2 (HER2) Detection by Electrochemical Biosensors

Department of Chemistry “Ugo Schiff”, University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino (FI), Italy
Author to whom correspondence should be addressed.
Academic Editors: Paolo Ugo and Ligia Moretto
Received: 26 July 2016 / Revised: 2 November 2016 / Accepted: 30 November 2016 / Published: 2 December 2016
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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In this study, we have designed and realized three simple electrochemical bioassays for the detection of the human epidermal growth factor receptor 2 (HER2) cancer biomarker using magnetic beads coupling screen-printed arrays. The different approaches were based on a sandwich format in which affibody (Af) or antibody (Ab) molecules were coupled respectively to streptavidin or protein A-modified magnetic beads. The bioreceptor-modified beads were used to capture the HER2 protein from the sample and sandwich assay was performed by adding the labeled secondary affibody or the antibody. An enzyme-amplified detection scheme based on the coupling of secondary biotinylated bioreceptor with streptavidin-alkaline phosphatase enzyme conjugate was then applied. The enzyme catalyzed the hydrolysis of the electro-inactive 1-naphthyl-phosphate to the electro-active 1-naphthol, which was detected by means of differential pulse voltammetry (DPV). Each developed assay has been studied and optimized. Furthermore, a thorough comparison of the analytical performances of developed assays was performed. Finally, preliminary experiments using serum samples spiked with HER2 protein were also carried out. View Full-Text
Keywords: affibody; immunosensor; magnetic beads; electrochemical detection; cancer biomarker affibody; immunosensor; magnetic beads; electrochemical detection; cancer biomarker

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Ilkhani, H.; Ravalli, A.; Marrazza, G. Design of an Affibody-Based Recognition Strategy for Human Epidermal Growth Factor Receptor 2 (HER2) Detection by Electrochemical Biosensors. Chemosensors 2016, 4, 23.

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