[Monoclonal antibodies to 8-oxo-2'-deoxyguanosine (8-hydroxyguanosine). Characteristics and use for determining DNA...

It has presently been established that the guanine base is one of the most sensitive and biologically significant target for the damaging action of active oxygen species on DNA, 7.8-dihydroxy-8-oxoguanine (8-oxoguanine, 8-hydroxyguanine) being the major degradation product and the most essential biomarker for DNA damage by active oxygen species. Murine monoclonal antibodies (MAbs) specifically recognizing 8-oxoguanine have been raised. Using competitive solid phase immunoenzymatic assay. (IEA) with peroxidase-antiperoxidase (PAP-method), a quantitative assay of this degradation product and characterization of affinity and specificity of MAbs have been carried out. The affinity constant (Kaff) Mabs for 8-oxo-2'-deoxyguanosine is equal to 1.3-10(6), that for 8-oxo-guanosine-to 1.10(6), exceeding by more than three orders of magnitude the Ka values for natural guanyl nucleosides and other possible cross-structure analogs. IEA was used to determine the DNA degradation product in gamma-irradiated DNA. The radiation-chemical yield of 8-oxoguanine (G = 0.57 molecules per 100 eV) is consistent with those obtained by other methods.