CHARACTERIZATION OF ANTIBODIES SPECIFIC FOR UV-DAMAGED DNA BY ELISA |
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Authors: | Guri Eggset Gunnar Volden Hans Krokan |
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Institution: | Institute of Medical Biology, University of Tromsø, N-9001 Tromsø, Norway;Institute of Clinical Medicine, University of Tromsø, N-9001 Tromsø, Norway;Norsk Hydro Research Centre, Section for Biotechnology, N-3901 Porsgrunn, Norway |
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Abstract: | Abstract— The specificity of affinity purified antibodies raised against UV-irradiated DNA was examined using an enzyme-linked immunosorbent assay. DNA irradiated with UV doses higher than needed for saturation with pyrimidine dimers bound increasing amounts of antibody. Photosensitized DNA, containing high amounts of pyrimidine dimers, showed very poor binding of antibody. When UV-irradiated DNA was given a second dose of 340-nm UV light, the binding of antibodies was abrogated. Taken together, this indicates a major specificity for (6-4)-photoproducts, which are photochemically reversed by UV light in the 340-nm region. The antibodies also showed little but detectable binding to pyrimidine glycols produced in DNA by oxidation with OsO4. Previously, we have used these antibodies for the detection of UV-induced DNA damage and its repair in human skin in vivo. These findings indicate that (6-4)-photoproducts, considered highly mutagenic, are repaired in human skin. |
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