Hvastkovs Eli G, So Minjeong, Krishnan Sadagopan, Bajrami Besnik, Tarun Maricar, Jansson Ingela, Schenkman John B, Rusling James F
Department of Chemistry, 55 North Eagleville Road, University of Connecticut, Storrs, Connecticut 06269, USA.
Anal Chem. 2007 Mar 1;79(5):1897-906. doi: 10.1021/ac061975q. Epub 2007 Jan 30.
Arrays suitable for genotoxicity screening are reported that generate metabolites from cytochrome P450 enzymes (CYPs) in thin-film spots. Array spots containing DNA, various human cyt P450s, and electrochemiluminescence (ECL) generating metallopolymer [Ru(bpy)2PVP10]2+ were exposed to H2O2 to activate the enzymes. ECL from all spots was visualized simultaneously using a CCD camera. Using benzo[a]pyrene as a test substrate, enzyme activity for producing DNA damage in the arrays was found in the order CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 > myoglobin, the same as the order of their metabolic activity. Thus, these arrays estimate the relative propensity of different enzymes to produce genotoxic metabolites. This is the first demonstration of ECL arrays for high-throughput in vitro genotoxicity screening.
据报道,适用于遗传毒性筛选的阵列可在薄膜斑点中由细胞色素P450酶(CYPs)产生代谢物。含有DNA、各种人细胞色素P450以及产生电化学发光(ECL)的金属聚合物[Ru(bpy)2PVP10]2+的阵列斑点被暴露于过氧化氢以激活这些酶。使用电荷耦合器件(CCD)相机同时观察所有斑点的ECL。以苯并[a]芘作为测试底物,发现在阵列中产生DNA损伤的酶活性顺序为CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 > 肌红蛋白,与其代谢活性顺序相同。因此,这些阵列可估计不同酶产生遗传毒性代谢物的相对倾向。这是首次证明用于高通量体外遗传毒性筛选的ECL阵列。