Open AccessA synthetic hapten for induction of thymine‐dimer‐specific antibodies
Author(s) -
KLOCKER Helmut,
AUER Bernhard,
BURTSCHER Helmut J.,
HIRSCHKAUFFMANN Monica,
SCHWEIGER Manfred
Publication year - 1984
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1984.tb08287.x
Subject(s) - hapten , radioimmunoassay , pyrimidine dimer , antiserum , chemistry , bovine serum albumin , dimer , thymine , cyclobutane , antibody , microbiology and biotechnology , ultraviolet light , antigen , conjugate , biochemistry , dna , biology , dna damage , photochemistry , immunology , organic chemistry , ring (chemistry) , mathematical analysis , mathematics
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 N 1 ‐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.