Assays for ROS-induced DNA damage

描述（由申请人提供）：该项目由ZeptoMetrix公司和Roswell Park癌症研究所合作。目的是开发灵敏、定量和简便的检测方法来测量活性氧（ROS）引起的DNA损伤。活性氧与衰老、癌症和几种神经系统疾病有关。将检测7种ros诱导的DNA损伤，即：嘧啶基甲氨基残基、8-氧- 7,9 -二氢鸟嘌呤（8-氧-鸟嘌呤）损伤、胸腺嘧啶乙二醇损伤、6-羟基-5,6-二氢胸腺嘧啶损伤、碱性损伤和两种双基损伤。32p后标记或质谱检测方法用于分析。ZeptoMetrix将使产品商业化，方便DNA测量的任何一种方法。产品将包括用于分析上述病变的参考化合物、载体和内部标准。32p后标记的载体提供了对感兴趣的DNA病变的明确识别。质谱测量的内部标准将同位素富集到比要定量的DNA损伤更高的质量。ZeptoMetrix预计这些产品最终最大的应用将是评估细胞和组织的修复能力。对修复研究有用的产品将包括参比化合物和含有已识别病变的双链DNA底物。利用电喷雾界面的液相色谱-质谱（LC-MS）将特别考虑到这一应用而开发。

DESCRIPTION (provided by applicant):This project is a collaboration between ZeptoMetrix Corporation and Roswell Park Cancer Institute. The objective is to develop sensitive, quantitative and facile assays for measuring DNA damage caused by reactive oxygen species (ROS). ROS have been implicated in aging, cancer and several neurological diseases. Seven ROS-induced DNA lesions will be assayed, namely: the formamido remnant of pyrimidine bases, the 8-oxo-7, 9-dihydroguanine (8-oxo-guanine) lesion, the thymine glycol lesion, the 6-hydroxy-5,6-dihydrothymine lesion, the abasic lesion and two double base lesions. 32P-postlabeling or mass spectroscopic methods of detection are employed with the assays. ZeptoMetrix will make products commercially available that facilitate DNA measurements by either method. Products will include reference compounds, carriers and internal standards for assaying the aforementioned lesions. Carriers for 32P-postlabeling provide unequivocal identification of the DNA lesion of interest. Internal standards for MS measurements will be isotopically enriched to a higher mass than the DNA lesion to be quantitated. ZeptoMetrix anticipates that eventually the largest application of these products will be for evaluating the repair capabilities of cells and tissues. Products useful for repair studies will include reference compounds and duplex DNA substrates containing identified lesions. Liquid Chromatography -Mass Spectrometry (LC-MS) utilizing an electrospray interface will be developed with this application especially in mind. PROPOSED COMMERCIAL APPLICATION: Reference materials, internal standards, and carriers for measuring oxidative DNA damage will be synthesized. These products will be offered commercially for facilitating DNA damage and repair by 32P-postlabeling or by LC-MS.