ABSTRACT
In this paper, we report the development of a sensitive label-free impedimetric biosensor based on the use of affibody as bioreceptor and gold nanostructured screen-printed graphite as a sensor platform for the detection of human epidermal growth factor receptor 2 (HER2). The affisensor is realized by immobilizing a terminal cysteine-modified affibody on gold nanoparticles. The sensor was characterized by electrochemical techniques and scanning electron microscopy (SEM). Furthermore, surface plasmon resonance (SPR) technology was also applied to explore the potential of affibodies as small-molecule discriminating tools. Using optimized experimental conditions, a single-use affisensor showed a good analytical performance for HER2 detection from 0 to 40 µg/L. The estimated limit of detection was 6.0 µg/L. Finally, the realized affisensor was applied to human serum samples.
Subject(s)
Biomarkers, Tumor/analysis , Biosensing Techniques/methods , Receptor, ErbB-2/analysis , Antibodies, Immobilized , Biomarkers, Tumor/blood , Biosensing Techniques/instrumentation , Electrochemical Techniques , Electrodes , Equipment Design , Gold/chemistry , Graphite/chemistry , Humans , Microscopy, Electron, Scanning , Nanoparticles/chemistry , Neoplasms/blood , Neoplasms/diagnosis , Receptor, ErbB-2/blood , Receptor, ErbB-2/immunology , Surface Plasmon ResonanceABSTRACT
A strategy for a label-free impedimetric immunosensor is described for detection of the textile dye Disperse Orange 1 (DO1). The compounds 1,12-diaminododecane (DADD) and then 1,7-diaminoheptane (DAH) were firstly successively grafted onto a glassy carbon electrode (GCE) surface by electro-oxidation of one amino group, while the other terminal amino group was modified with the antibody anti-DO1. The construction process of the immunosensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy and capacitance measurements. The electron transfer resistance (Rct) exhibited an effective response to the affinity between the immobilized antibody and the antigen in solution. The linear range for the target compound was from 5.0 nmol L(-1) to 0.5 µmol L(-1) (R=0.9980), and the limit of detection (LOD) was 7.56 nmol L(-1). The proposed impedimetric immunosensor has the advantages of simplicity, cost-effectiveness, and sensitivity.
