Direct electron transfer bioelectronic interfaces: application to clinical analysis.
Bioelectronic interfaces based on direct electron transfer to proteins and enzymes immobilised at functional electrode surfaces are currently under development and the potential of two such systems for application to clinical measurement will be outlined. The first is the detection of free radical production via direct electrochemistry of cytochrome c immobilised covalently at modified gold electrodes. The redox protein cytochrome c has been immobilised covalently to gold electrodes surface-modified with N-acetyl cysteine via carbodiimide condensation. The electrodes thus produced were used to measure directly the enzymatic and cellular production of the superoxide anion radical (O2(-). The superoxide radical reduced the immobilised cytochrome c which was immediately re-oxidised by the surface-modified gold electrode poised at a potential of +25 mV (vs Ag/AgCl). The electron transfer rate constant (ket) of this process was 3.4 +/- 1.2 s(-1). The rate of current generation was directly proportional to the rate of O2(-) production. The essentially reagentless system produced was designed to be applied ultimately to continuous monitoring of free radical activity in vivo since there is evidence that oxygen-derived free radical species act as mediators which cause and perpetuate inflammation in disease states, including rheumatoid arthritis and neurodegenerative disorders. The second systems are pseudo-homogeneous immunoassays based on direct electron transfer to horseradish peroxidase. Horseradish peroxidase enzyme electrodes based on activated carbon (HRP-ACE) have been constructed by simple passive adsorption.(ABSTRACT TRUNCATED AT 250 WORDS)
McNeil
C J
CJ
Department of Clinical Biochemistry, Medical School, University of Newcastle upon Tyne, UK.
Athey
D
D
Ho
W O
WO
D003574:Cytochrome c Group / Q000737:chemistry*; D004563:Electrochemistry; D004566:Electrodes; D004579:Electron Transport; D004582:Electronics, Medical; D004800:Enzymes, Immobilized; D005609:Free Radicals; D006046:Gold; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay; D009504:Neutrophils / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D013481:Superoxides / Q000378:metabolism
D003574:Cytochrome c Group; D004800:Enzymes, Immobilized; D005609:Free Radicals; D013481:Superoxides; D006046:Gold; D006735:Horseradish Peroxidase