Triggering the redox reaction of cytochrome c on a biomimetic layer and elimination of interferences for NADH detection.
Biomimetic layers triggering the redox process of cytochrome c (cyt c) by beta-nicotinamide adenine dinucleotide (NADH) were fabricated and applied for the detection of NADH. A probe was constructed based on a conducting polymer (poly-5,2':5',2''-terthiophene-3'-carboxylic acid, poly-TTCA) formed on the Au nanoparticles, which were deposited on a screen-printed carbon electrode; the probe was modified with biomaterials including cyt c, lipids (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cardiolipin), and ubiquinone, which are involved in electron transfer sequence in the cell membrane. To eliminate affection of foreign biological species, we assembled a lipid bilayer using the Langmuir-Blodgett technique by controlling the density of the outward lipid layer. The characteristics of the biomimetic layers were investigated by cyclic voltammetry, impedance spectrometry, quartz crystal microbalance, and X-ray photoelectron spectroscopy. The electron transfer process triggered by the presence of NADH was used to determine the NADH concentration. L-ascorbic acid, uric acid, and NADPH did not show any affection during the detection of NADH at the density controlled-lipid bilayer. Real sample analysis was successfully performed to evaluate the reliability of the proposed biomimetic probe.
Lee
Kyung-Sun
KS
Department of Chemistry, Pusan National University, Busan, South Korea.
Won
Mi-Sook
MS
Noh
Hui-Bog
HB
Shim
Yoon-Bo
YB
D000818:Animals; D032701:Biomimetics; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D045304:Cytochromes c / Q000737:chemistry*; D055664:Electrochemical Techniques; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D006736:Horses; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008055:Lipids / Q000737:chemistry; D008422:Materials Testing; D053768:Metal Nanoparticles / Q000737:chemistry; D008958:Models, Molecular; D015394:Molecular Structure; D009243:NAD / Q000032:analysis*; D010084:Oxidation-Reduction; D011487:Protein Conformation
D008051:Lipid Bilayers; D008055:Lipids; D009243:NAD; D006046:Gold; D045304:Cytochromes c