In vivo detection of glutathione disulfide and oxidative stress monitoring using a biosensor.
A highly sensitive in vivo biosensor for glutathione disulfide (GSSG) is developed using covalently immobilized-glutathione reductase (GR) and -β-nicotinamide adenine dinucleotide phosphate (NADPH) on gold nanoparticles deposited on poly[2,2':5',2″-terthiophene-3'-(p-benzoic acid)] (polyTTBA). The fabricated biosensor was characterized with SEM, TEM, XPS, and QCM. Analytical parameters affecting the biosensor performance were optimized in terms of applied potential, NADPH:GR ratio, temperature, and pH. A linear calibration plot is obtained using chronoamperometry in the dynamic range between 0.1 μM and 2.5 mM of GSSG, with a detection limit of 12.5 ± 0.5 nM. The developed biosensor is applied to detect GSSG in a real plasma sample. A microbiosensor was applied to detect the in vivo GSSG concentration to monitor the oxidative stress caused by diquat and t-butyl hydroperoxide. The results obtained are reliable, implying a promising approach for a GSSG biosensor in clinical diagnostics and oxidative stress monitoring.
Department of Chemistry and Institute of BioPhysio Sensor Technology, Pusan National University, Busan 609-735, Republic of Korea.
Hui-Bog
HB
Noh
Pranjal
P
Chandra
Jeon Ok
JO
Moon
Yoon-Bo
YB
Shim
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D002244:Carbon; D055664:Electrochemical Techniques; D004566:Electrodes; D005898:Glass; D019803:Glutathione Disulfide / Q000097:blood*; D005980:Glutathione Reductase / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008297:Male; D053768:Metal Nanoparticles / Q000648:ultrastructure; D009249:NADP / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D013499:Surface Properties / Q000187:drug effects; D013876:Thiophenes / Q000494:pharmacology
D013876:Thiophenes; C019101:alpha-terthienyl; D009249:NADP; D002244:Carbon; D006046:Gold; D005980:Glutathione Reductase; D019803:Glutathione Disulfide