Monoclonal antibodies to DNA modified by the carcinogen N-acetoxy-N-2 acetyl aminofluorene.

BALB/c mice were immunized against acetyl aminofluorene-substituted guanosine (AAF.Guo). Hybridomas were generated from the immune splenocytes fused with the SP2/0.Ag8 cell line. Four monoclonal antibodies (mAb) were selected. Two of them recognize mainly AAF. substituted DNA but also native DNA, while the others (derived from the hybridoma 16) are specific for the modified DNA. Using a solid phase radioimmunoassay, the fine specificity of mAb 16 was examined. The interaction between purified mAb 16 an AAF.DNA is inhibited by either AAF.GMP, AAF.Guo, or AAF.DNA. Moreover this interaction is slightly inhibited by aminofluorene-substituted guanosine (AF.Guo). By control, free AAF, fluorenamine, guanosine, desoxy GMP and native DNA do not compete in the assay. This mAb 16, which specifically recognizes the AAF-nucleotide complex may be a very useful tool to analyze the carcinogenic processes. Such an antibody can also be used for chromosomal localization of genes by AAF-substituted probes.