A third-generation biosensor for hydrogen peroxide based on the immobilization of horseradish peroxidase on a disposable carbon nanotubes modified screen–printed electrode

Abstract

A screen-printed carbon nanotube (CNTs) based disposable third generation biosensor for hydrogen peroxide (H 2 O 2 ) was constructed by mixing CNTs with a cellulose acetate binder on an epoxy substrate. The surface was covered with a layer consisting of horseradish peroxidase (HRP) crosslinked to bovine serum albumin with glutaraldehyde. The CNTs acted as both the electrode material and charge promoter to facilitate the direct electron transfer between immobilized HRP and the modified electrode. At a working potential of −300 mV (vs. Ag/AgCl), the biosensor displays an excellent electrocatalytic response to the reduction of H 2 O 2 without the aid of a mediator. It has a fast (3 s) amperometric response and a linear calibration in the concentration range 0.005-0.1 mmol L −1 , with a 0.85 μM detection limit (at an S/ N of 3). The biosensor can be easily produced, is stable and reproducible.

A simple and sensitive hydrogen peroxide detection with horseradish peroxidase immobilized on pyrene modified acid‐treated single‐walled carbon nanotubes

“…In some cases where enzyme–electrode interactions are suitably adjusted, it is possible to achieve direct electron transfer (DET) between the enzyme and the electrode. Horseradish peroxidase (HRP) is one of the most widely used enzymes to construct H 2 O 2 biosensors as it was shown to achieve DET when used with suitable electrode confirmation (third‐generation biosensors) …”

“…Furthermore, SWCNTs have been utilized as an enzyme immobilization matrix and shown to enhance electron transfer efficiency . The most widely utilized HRP‐electrode confirmations based on carbon nano‐materials are glutaraldehyde crosslinking, layered nano‐orientation using covalent attachment and electrostatic interactions . Each method has its challenge and its advantages depending on the biomolecule used.…”

“…Horseradish peroxidase (HRP) is one of the most widely used enzymes to construct H 2 O 2 biosensors as it was shown to achieve DET when used with suitable electrode confirmation (third-generation biosensors). 12,13 Recently, several different approaches have been developed to construct mediator-free enzymatic H 2 O 2 biosensors using the Corrections added on 25 February 2020, after first online publication: the result section in the abstract has been modified. Also the supporting information, Figure 1 and the figure citations in the section "Electrochemical performance of H 2 O 2 biosensor" has been changed.…”

Carbon-black-doped Polyimide-modified Glassy Carbon Electrode for Sensitive Nonenzymatic Amperometric Determination of Hydrogen Peroxide

“…(2) Many analytical methods have been proposed for the detection of H 2 O 2 such as chromatography, (3) chemiluminescence, (4) and electrochemistry. (5)(6)(7) Electrochemistry has proved to be effective for H 2 O 2 determination. Redox proteins including horseradish peroxidase, (8,9) myoglobin, (10) hemoglobin, (11) and cytochrome c (12) have been used for the fabrication of H 2 O 2 biosensors.…”

Novel Enzymatic Rhodium Modified Poly(styrene‐g‐oleic amide) Film Electrode for Hydrogen Peroxide Detection

“…Horseradish peroxidase a heme containing glycoprotein extracted from horseradish root is the most commonly used peroxidase in many analytical applications . There have been an interesting attention on the third generation hydrogen peroxide biosensor based on direct electron transfer between electrode and immobilized peroxidase . Even though, these studies have been useful for the investigation of enzyme structure, redox transformation mechanisms of protein molecules and metabolic processes, it is usually difficult to achieve the direct electron transfer between the enzyme and bare electrode owing to the deeply embedded redox active center and unfavorable orientation of enzymes.…”