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.
