| RRC ID |
38164
|
| Author |
Inoue KY, Ino K, Shiku H, Kasai S, Yasukawa T, Mizutani F, Matsue T.
|
| Title |
Electrochemical monitoring of hydrogen peroxide released from leucocytes on horseradish peroxidase redox polymer coated electrode chip.
|
| Journal |
Biosens Bioelectron
|
| Abstract |
We developed an electrochemical-sensing device for continuous monitoring extracellular hydrogen peroxide (H(2)O(2)). The device consists of an indium-tin-oxide electrode coated with osmium-polyvinylpyridine gel polymer containing horseradish peroxidase (Os-HRP) and a poly-dimethyl siloxane well to house the cells on the chip. Granulocyte-like differentiated HL-60 cells were accommodated in the well and stimulated with phorbol 12-myristate 13-acetate (PMA), which triggered the generation of H(2)O(2). The extracellular H(2)O(2) released from the cells was enzymatically reduced at the Os-HRP-modified electrode chip using Os(II) as an electron donor, resulting in reduction current responses by the device. The reduction current increased immediately upon PMA stimulation and this current transient was similar to that obtained by conventional chemiluminescence assays using sodium luminol. Apocynin, an inhibitor of NADPH oxidase activation, eliminated both the electrochemical and chemiluminescence signals. On the other hand, superoxide dismutase (SOD) increased the amperometric signals and catalase (CAT) decreased, whereas SOD decreased luminescence emission and CAT did not. These results were in accordance with the expected reaction mechanism, and strongly indicate that this new electrochemical-sensing device successfully detects extracellular H(2)O(2) production.
|
| Volume |
25(7)
|
| Pages |
1723-8
|
| Published |
2010-3-15
|
| DOI |
10.1016/j.bios.2009.12.014
|
| PII |
S0956-5663(09)00665-4
|
| PMID |
20060284
|
| MeSH |
Biological Assay / instrumentation*
Biosensing Techniques / instrumentation*
Biosensing Techniques / methods
Cell Line
Coated Materials, Biocompatible / chemistry
Conductometry / instrumentation*
Conductometry / methods
Electrodes*
Enzymes, Immobilized / chemistry
Equipment Design
Equipment Failure Analysis
HL-60 Cells
Horseradish Peroxidase / chemistry*
Humans
Leukocytes / metabolism*
Oxidation-Reduction
Polymers / chemistry*
Reproducibility of Results
Sensitivity and Specificity
|
| IF |
10.257
|
| Times Cited |
20
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
CHEMISTRY, ANALYTICAL
BIOPHYSICS
NANOSCIENCE & NANOTECHNOLOGY
ELECTROCHEMISTRY
|
| Resource |
| Human and Animal Cells |
|
