Department of Chemistry, Pusan National University, Busan, South Korea.
Biomaterials. 2010 Oct;31(30):7827-35. doi: 10.1016/j.biomaterials.2010.06.052. Epub 2010 Jul 24.
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.
仿生层通过β-烟酰胺腺嘌呤二核苷酸（NADH）触发细胞色素 c（cyt c）的氧化还原过程，被制备并应用于 NADH 的检测。基于在金纳米粒子上沉积的导电聚合物（聚 5,2':5',2''-三噻吩-3'-羧酸，聚 TTCA）构建了一个探针；该探针用包括细胞色素 c、脂质（1,2-二油酰基-sn-甘油-3-磷酸乙醇胺（DOPE）和心磷脂）和泛醌在内的生物材料进行了修饰，这些物质参与了细胞膜中的电子转移序列。为了消除外源生物物质的影响，我们使用 Langmuir-Blodgett 技术通过控制外向脂质层的密度组装了一个脂质双层。通过循环伏安法、阻抗谱、石英晶体微天平以及 X 射线光电子能谱研究了仿生层的特性。通过存在 NADH 触发的电子转移过程来确定 NADH 的浓度。在控制密度的脂质双层中检测 NADH 时，L-抗坏血酸、尿酸和 NADPH 没有显示出任何影响。成功进行了实际样品分析，以评估所提出的仿生探针的可靠性。
