A synthetic hapten for induction of thymine-dimer-specific antibodies.
High specificity and sensitivity of thymine cyclobutane dimer (thy[]thy) detection were obtained by a radioimmunoassay. Attempts to raise thy[]thy-monospecific antibodies with antigens produced according to conventional methods were unsuccessful. Thy[]thy-specific antibodies could only be raised by using a new strategy to bind thy[]thy to protein: thymine was activated by trimethylsilylation and alkylated at N1 yielding N1-thyminebutanoic acid which was dimerised by ultraviolet treatment. The resulting derivative of thymine cyclobutane dimer was coupled to bovine serum albumin by the active-ester method. The new strategy appears to be generally applicable for binding haptens, such as DNA bases, photoproducts etc, to proteins via a derivative containing a carboxyl group. Immunisation of rabbits with the thy[]thy-bovine-serum-albumin conjugate prepared by the new method resulted in a highly specific antiserum which allows detection of thy[]thy down to 0.06 p mol (15pg). The thy[]thy-specific radioimmunoassay was applied to measure thy[]thy formed in human fibroblasts which were exposed to sunlight at altitudes of 600 m or 2300 m. The amounts of thy[]thy formed in an hour corresponded to doses of 14 J m-2 and 24 J m-2, respectively, of an ultraviolet light lamp emitting predominantly 245-nm light.
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA / Q000032:analysis / Q000528:radiation effects; D005347:Fibroblasts / Q000032:analysis; D006241:Haptens / Q000276:immunology; D006801:Humans; D011740:Pyrimidine Dimers / Q000032:analysis / Q000276:immunology*; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine / Q000276:immunology; D014466:Ultraviolet Rays
D000906:Antibodies; D000941:Antigens; D006241:Haptens; D011740:Pyrimidine Dimers; D012710:Serum Albumin, Bovine; D004247:DNA