ACETOXY-ACETYLAMINOFLUORENE-MODIFIED DGTP CAN BE USED TO LABEL OLIGONUCLEOTIDES OR DNA ENZYMATICALLY

A competitive enzyme immunoassay using a bispecific monoclonal antibod y (Bi-MAb) was developed to quantify acetylaminofluorene (AAF) adducts fixed on DNA and then compared to the spectrophotometric method. It w as shown that this simple method allowed the measurement of as low as 2 pmol per assay of AAF bound to DNA. This technique was used to monit or synthesis and purification of N-acetoxy-N-2-acetylaminofluorene mod ified dGTP (AAF-dGTP). It was shown that AAF-dGTP can be a substrate f or the terminal deoxynucleotidyl transferase. Finally, using the Bi-MA b we developed a non-radioactive sandwich hybridization assay making u se of oligonucleotide covalently bound to microwells and of synthetic oligonucleotide tailed with AAF-dGTP as a probe. (C) 1997 Academic Pre ss Limited.