Development and Application of Novel Biomarkers for Regulation of Free Radical Reactions in Biological Systems

氧化应激可能会引起自由基反应，在膜、蛋白质、酶和DNA中产生有害的修饰。与年龄相关的疾病，如癌症、动脉粥样硬化和糖尿病，被认为与氧化应激有关，尽管具体机制仍不清楚。我们的研究小组一直致力于通过应用免疫化学来开发新的ELISA法。我们研制了多种针对氧化损伤的DNA碱基的单抗和多克隆抗体，如8-羟基脱氧鸟苷和脂质过氧化产物丙二醛、4-羟基壬烯醛和环烯丙氨酸等。我们还参与了亚油酸过氧化氢氧化修饰蛋白质的分离和结构测定。通过化学研究，我们确定了针对早期脂质过氧化产物的表位结构为己酰赖氨酸类化合物。我们成功地获得了针对己酰基赖氨酸结构的单抗和多克隆抗体作为表位。最近，我们还获得了针对Dityroine的多克隆抗体，作为Myropexidase氧化损伤蛋白之一的新的生物标志物。利用这些单抗和多克隆抗体，我们正在尝试建立ELISA法。

Oxidative stress may cause free radical reactions to produce deleterious modifications in membranes, proteins, enzymes and DNA. Age-related diseases such as cancer, atherosclerosis and diabetes are supposed to be correlated with oxidative stress although the detailed mechanisms are still unclear. Our research group has been involved in developing novel ELISA methods by application of immnochemistry. We have developed many types of monoclonal and polyclonal antibodies which are specific to oxidatively damaged DNA base such as 8-OH-deoxyguanosine and lipid peroxidation products including malondialdehyde, 4-hydroxynonenal and aclorein etc. We have also been involved in isolation and structure determination of oxidatively modified proteins by linoleic acid hydroperoxide. By the chemical investigation, we have determined the structure of epitope which is specific to early stage lipid peroxidation products as hexanoyl lysine type compounds. We have succeeded in obtaining both monoclonal and polyclonal antibodies specific for hexanoyl lisine structure as epitope. Recently, we also obtained polyclonal antibody specific to dityrosine as the new biomarker for one of the oxidatively damaged proteins by myropexidase. Using these monoclonal and polyclonal antibodies, we are trying to establish ELISA methods.