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

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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.