Further characterisation of a polyclonal antiserum for DNA photoproducts: the use of different labelled antigens to control its specificity.
Synthetic polynucleotides irradiated with far (254 nm) or near (320 nm) UV-light were used to characterise 3 different radioimmunoassay systems. Antiserum raised against DNA irradiated with a high dose of far-UV-light was found to have at least 2 antibody populations. A competitive assay in which the labelled antigen was irradiated at 254 nm was found to be specific for Pyr(6-4)Pyo adducts, the antibody-binding sites being sensitive to a secondary photolytic dose of 320-nm light. When the labelled antigen was irradiated with 320-nm light the assay was specific for cyclobutane dimers. This assay had the same specificity as one consisting of labelled DNA irradiated with 254-nm light and an antiserum raised against DNA irradiated at 320 nm in the presence of acetophenone. These assay systems were used to demonstrate the dose-dependence of the induction and photolytic degradation of Pyr(6-4)Pyo adducts by a near-UV-light source.
D L
DL
Mitchell
C A
CA
Haipek
J M
JM
Clarkson
D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D000918:Antibody Specificity; D002417:Cattle; D000939:Epitopes / Q000276:immunology / Q000528:radiation effects; D007106:Immune Sera; D011067:Poly dA-dT / Q000276:immunology / Q000528:radiation effects*; D011089:Polydeoxyribonucleotides / Q000528:radiation effects*; D011740:Pyrimidine Dimers / Q000276:immunology; D011863:Radioimmunoassay; D014466:Ultraviolet Rays
D000974:Antibodies, Antinuclear; D000939:Epitopes; D007106:Immune Sera; D011089:Polydeoxyribonucleotides; D011740:Pyrimidine Dimers; D011067:Poly dA-dT