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1.
J Mol Recognit ; 34(11): e2928, 2021 11.
Article in English | MEDLINE | ID: mdl-34378255

ABSTRACT

In this study, an innovative strategy was proposed for the electrocatalytical reduction and enzymatic biosensing of hydrogen peroxide (H2 O2 ) using chronoamperometry technique. For the first time, immobilization of horseradish peroxidase (HRP) in polydopamine-modified magnetic nanoparticles (PDA-MNPs) was successfully performed. Also, poly(l-arginine/toluidine blue) film-modified glassy carbon electrode was constructed through co-electropolymerization of l-arginine and toluidine blue on the surface of GCE using cyclic voltammetry technique. The engineered hybrid thin film provides strong functionalities for efficient grafting of PDA-MNPs which, in turn, enable the covalent immobilization of HRP. The proposed biosensor was used for the detection of H2 O2 in the range of 0.5-30 µM with a low limit of quantification 0.23 µM. It also was successfully applied for the investigation of hydrogen peroxide in human plasma samples.


Subject(s)
Biosensing Techniques/methods , Enzymes, Immobilized/metabolism , Ferric Compounds/chemistry , Horseradish Peroxidase/metabolism , Hydrogen Peroxide/blood , Indoles/chemistry , Metal Nanoparticles/chemistry , Polymers/chemistry , Arginine/chemistry , Electrochemistry , Electrodes , Enzymes, Immobilized/chemistry , Horseradish Peroxidase/chemistry , Humans , Tolonium Chloride/chemistry
2.
J Mol Recognit ; 34(5): e2884, 2021 05.
Article in English | MEDLINE | ID: mdl-33393155

ABSTRACT

In this study, a selective and sensitive molecular imprinting-based electrochemical sensors, for horseradish peroxidase (HRP) entrapment was fabricated using electro polymerization of ß-Cyclodextrin (ß-CD) on the surface of glassy carbon electrode. Poly beta-cyclodextrin P(ß-CD) provide efficient surface area for self-immobilization of HRP as well as improve imprinting efficiency. The proposed imprinted biosensor successfully utilized for detection of HRP with excellent analytical results which linear range is 0.1 mg/mL to 10 ng/mL with LOD of 2.23 ng/mL. Furthermore, electrocatalytical activity of the prepared biosensor toward the reduction of hydrogen peroxide was investigated in the ranges of 1 to 15 µM with a detection limit of 0.4 µM by using chronoamperometry technique. The developed biosensor was used for the detection of hydrogen peroxide in unprocessed human plasma sample.


Subject(s)
Horseradish Peroxidase/chemistry , Hydrogen Peroxide/blood , Molecularly Imprinted Polymers/chemistry , beta-Cyclodextrins/chemistry , Biosensing Techniques , Blood Chemical Analysis , Enzymes, Immobilized/chemistry , Humans , Microscopy, Electron, Scanning
3.
Int J Biol Macromol ; 145: 311-324, 2020 Feb 15.
Article in English | MEDLINE | ID: mdl-31881306

ABSTRACT

A low cost, sensitive and selective electrochemical imprinted biosensor for horseradish peroxidase-conjugated prostate-specific antigen (PSA) was designed and prepared based on the combination of Toluidine blue (TB) and self-assembly surface molecular imprinting technique. Poly toluidine blue [P(TB)] provided high surface area for dense loading of HRP-PSA antibody on GCE surface. P(TB), as supporting material, could effectively enhance imprinting efficiency and the electrode conductivity and facilitate electron transfer. The imprinted biosensor was characterized through Field emission scanning electron microscopy (FE-SEM), Energy-dispersive X-ray spectroscopy (EDX) and electrochemical methods. The proposed biosensor indicates very highly electrocatalytical activity for the reduction of hydrogen peroxide (H2O2). Also, engineered biosensor was used for determination of H2O2 by different electrochemical techniques including differential pulse voltammetry, square wave voltammetry and chronoamperometry. Under the optimized conditions, the proposed bio-imprinted polymer exhibit excellent electrocatalytical activity toward the reduction of H2O2 with wide linear range of 0.001 to 40 mM and a low limit of quantification (LLOQ) of 1 µM.


Subject(s)
Acrylic Resins/chemistry , Biological Assay , Biosensing Techniques , Horseradish Peroxidase/chemistry , Hydrogen Peroxide/blood , Prostate-Specific Antigen/chemistry , Tolonium Chloride/analogs & derivatives , Antibodies/chemistry , Electrochemical Techniques , Electrodes , Enzymes, Immobilized/chemistry , Humans , Limit of Detection , Male , Molecular Imprinting/methods , Oxidation-Reduction , Prostate/metabolism , Prostate-Specific Antigen/blood , Tolonium Chloride/chemistry
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