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The Use of a β-lactamase-based Conductimetric Biosensor Assay to Detect Biomolecular Interactions
Published on: February 1, 2018
Polythiophene-g-poly(ethylene glycol) with Lateral Amino Groups as a Novel Matrix for Biosensor Construction
Huseyin Akbulut1, Guliz Bozokalfa2, Duygu N Asker2
1Istanbul Technical University , Department of Chemistry, Faculty of Science and Letters, Istanbul, Turkey.
Abstract:
In the ever-expanding field of conducting polymer research, functionalized graft hybrid copolymers have gained considerable interest in the biomedical engineering and biosensing applications, particularly. In the present work, a new biosensor based on conducting graft copolymer for the detection of phenolic compounds was developed. Thereby, a robust and novel material, namely "polythiophene-g-poly(ethylene glycol) with lateral amino groups" (PT-NH2-g-PEG) hybrid conducting polymer was synthesized via Suzuki condensation polymerization and characterized with (1)H NMR analysis, UV-vis spectroscopy, gel permeation chromatography (GPC) and fluorescence spectroscopy. PT-NH2-g-PEG architecture was then applied as an immobilization matrix to accomplish extended biosensing function. In a typical process, Laccase was utilized as a model enzyme for the detection of phenolic compounds. Detailed surface characterization of PT-NH2-g-PEG/Lac was performed by cyclic voltammetry, electrochemical impedance spectroscopy, atomic force microscopy, fluorescence microscopy and scanning electron microscopy measurements. Optimum pH and polymer amount were found to be pH 6.5 and 0.5 mg polymer, respectively, with the linear range of 0.0025-0.05 mM and 132.45 μA/mM sensitivity. The kinetic parameters of PT-NH2-g-PEG/Lac are 0.026 mM for Km(app) and 7.38 μA for Imax, respectively. Furthermore, the PT-NH2-g-PEG/Lac biofilm was retained 82% of its activity for 12 days indicating excellent recovery as tested with artificial wastewater.

