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Computational approach and electrochemical measurements for protein detection with MIP-based sensor
Abstract : Rapid and accurate detection of proteins in biological fluids is increasingly required in the biomedical environment. Actually, it is performed with conventional techniques, which are generally run by robotized platforms at centralized laboratories. In this work, molecular dynamics calculations and an experimental procedure were conducted to set up electrochemical sensors based on polypyrrol (PPy) molecular imprinted polymers (MIP) for proteins detection. Here, prostate-specific antigen (PSA) was selected as a template model. Computational calculations indicate that for any PPy conformation and any amino-acid location in the protein, PSA molecules remain strongly inserted in the PPy polymer without biological alterations. One from possible orientations, appeared to be most probable as it presents the lowest absorption energy (-363 kcal mol-1) and largest contact area (4034.1 Å2). The device was then elaborated by in situ electropolymerization of PPy films. MIP's thickness and extraction duration were optimized by chronoamperometry. Square wave voltammetry technique was investigated for PSA detection in standard solution in the concentration range of 3x10 -8 ng.ml-1- 300 ng ml-1. According to the Hill equation, the equilibrium dissociation constant Kdbetween PSA and its imprint was estimated at Kd = (1.02 ± 0.54) × 10-14 M, confirming the strong binding between the designed MIP and the protein as predicted by the computational study. PSA concentration values directly measured in 35 human serum samples were found closely correlated to those measured by the ELISA technique. The promising fast and low-cost sensor might be used successfully for proteins detection at low concentrations with high selectivity and reproducibility.
https://hal-univ-lemans.archives-ouvertes.fr/hal-02464043
Contributeur : Nourdin Yaakoubi Connectez-vous pour contacter le contributeur
Soumis le : dimanche 2 février 2020 - 20:18:11
Dernière modification le : mercredi 3 novembre 2021 - 05:30:22
Contributeur : Nourdin Yaakoubi Connectez-vous pour contacter le contributeur
Soumis le : dimanche 2 février 2020 - 20:18:11
Dernière modification le : mercredi 3 novembre 2021 - 05:30:22
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