The present invention relates to methods and materials for the detection and quantitation 8-OH-Ade in biological specimens. Specifically, the present invention is directed to a group of highly specific
monoclonal antibodies reactive with the modified
nucleoside structure 8-OH-Ade, and to various immunoassays for 8-OH-Ade utilizing these
monoclonal antibodies. The
monoclonal antibodies of the present invention may be used in assays for diagnosing or monitoring the progression of certain types of
cancer, in addition to a variety of other diseases associated with
mutagenesis resulting from
oxidative damage of
DNA. Assays utilizing the monoclonal antibodies of the present invention may also be used to analyze or monitor
toxicant exposure, such as from environmental sources. The monoclonal antibodies of the present invention were prepared with the
immunogen 8-OH-
adenosine coupled to
keyhole limpet
hemocyanin (KLH), not to 8-OH-Ade directly. It is believed that the monoclonal antibodies bind with the base portion of the structure (8-OH-Ade) and not the
carbohydrate (
ribose) or
protein linkage region of the conjugate, because, as demonstrated, conjugates bound to nucleosides other than 8-OH-
adenosine were unreactive with these antibodies. Therefore, the antibodies of the present invention can be used to detect and quantitate (by the use of a
standard curve) the presence of 8-OH-Ade in biological specimens of
DNA. Procedures for such an
assay include immobilizing the
DNA, denaturing it to disrupt the base-
pairing scheme exposing the
free base structures, and quantitating the amount of 8-OH-Ade present per amount of DAN in a quantitative
immunoassay.