So Minjeong, Hvastkovs Eli G, Schenkman John B, Rusling James F
Department of Chemistry, University of Connecticut, Storrs, CT 06269-3060, United States.
Biosens Bioelectron. 2007 Nov 30;23(4):492-8. doi: 10.1016/j.bios.2007.06.017. Epub 2007 Jul 28.
Simultaneous optical and voltammetric detection of bioactivated genotoxicity is reported for the first time employing ultrathin films of DNA, model metabolic enzymes, and electrochemiluminescence (ECL) generating metallopolymer [Ru(bpy)2PVP10]2+ on pyrolytic graphite (PG) electrodes. Cytochrome P450cam and myoglobin were used as model monoxygenase enzymes to mimic in vivo processes. Sensor film growth and component amounts were monitored using a quartz crystal microbalance (QCM). Subsequent to the enzyme reaction, DNA damage in the sensor films was measured simultaneously using a simple apparatus combining a standard voltammetry cell coupled with an optical fiber and photomultiplier tube. The model enzyme reaction converted styrene to styrene oxide, which reacts with DNA nucleobases. ECL and SWV signals increased with enzyme reaction time on the scale of several min, and provided relative enzyme turnover rates for DNA damage suitable for toxicity screening applications. Within 1 min, the sensor detects approximately 3 damaged bases per 10,000 DNA bases using this simultaneous detection.
首次报道了利用DNA超薄膜、模型代谢酶以及在热解石墨（PG）电极上生成电化学发光（ECL）的金属聚合物[Ru(bpy)₂PVP₁₀]²⁺，同时进行光学和伏安法检测生物活化的遗传毒性。细胞色素P450cam和肌红蛋白被用作模型单加氧酶来模拟体内过程。使用石英晶体微天平（QCM）监测传感器膜的生长和组分含量。酶反应后，使用一个简单的装置同时测量传感器膜中的DNA损伤，该装置将标准伏安法电池与光纤和光电倍增管相结合。模型酶反应将苯乙烯转化为环氧苯乙烷，后者与DNA碱基发生反应。ECL和方波伏安法（SWV）信号在几分钟的时间尺度上随酶反应时间增加，并提供了适用于毒性筛选应用的DNA损伤相对酶周转速率。使用这种同时检测方法，传感器在1分钟内可检测出每10000个DNA碱基中约3个受损碱基。
