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A label-free impedimetric immunosensor based on covalent immobilization of anti-E. Coli antibody via a copper-catalyzed azide-alkyne cycloaddition reaction
- Source :
- Analytical and Bioanalytical Chemistry. 412:5077-5087
- Publication Year :
- 2020
- Publisher :
- Springer Science and Business Media LLC, 2020.
-
Abstract
- In this article, an original method is proposed for address and covalent immobilization of anti-E. coli antibodies on a screen-printed electrode (SPE). The method is based on a copper-catalyzed “click” reaction between a polyvinylbenzylazide (PVBA) film electrochemically deposited on the electrode surface and acetylene fragments of propargyl-N-hydroxysuccinimide ester. The products of electrochemical oxidation of copper particles incorporated in the polymer film on the electrode were first used for catalysis of the click reaction. This approach allowed us to reduce the immobilization time from a few hours for conventional methods to just 30 min, and to prevent denaturation of the immunoreceptor. The modified electrodes were characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV). Based on the results obtained, a label-free impedimetric immunosensor for E. coli detection was developed. The detection limit of the immunosensor was estimated as 6.3 CFU/ml, with a linear range of 103–106 CFU/ml. The immunosensor demonstrated good stability during 30 days of storage in phosphate buffer solution (PBS, pH 7) and selectivity toward excess Staphylococcus aureus bacteria.
- Subjects :
- Azides
Staphylococcus aureus
Metal Nanoparticles
02 engineering and technology
Electrochemistry
01 natural sciences
Biochemistry
Catalysis
Analytical Chemistry
chemistry.chemical_compound
Limit of Detection
Escherichia coli
Electrodes
Immunoassay
Detection limit
Cycloaddition Reaction
Chemistry
010401 analytical chemistry
021001 nanoscience & nanotechnology
Antibodies, Bacterial
Cycloaddition
0104 chemical sciences
Dielectric spectroscopy
Covalent bond
Alkynes
Dielectric Spectroscopy
Click chemistry
Azide
Cyclic voltammetry
0210 nano-technology
Antibodies, Immobilized
Copper
Nuclear chemistry
Subjects
Details
- ISSN :
- 16182650 and 16182642
- Volume :
- 412
- Database :
- OpenAIRE
- Journal :
- Analytical and Bioanalytical Chemistry
- Accession number :
- edsair.doi.dedup.....ab097effa314d14833b6245a5eb4f8e1