Further characterisation of a polyclonal antiserum for DNA photoproducts: the use of different labelled antigens to control its specificity

Mutat Res. 1985 Sep;146(2):129-33. doi: 10.1016/0167-8817(85)90002-1.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Antinuclear / immunology*
  • Antibody Specificity
  • Cattle
  • Epitopes / immunology
  • Epitopes / radiation effects
  • Immune Sera
  • Poly dA-dT / immunology
  • Poly dA-dT / radiation effects*
  • Polydeoxyribonucleotides / radiation effects*
  • Pyrimidine Dimers / immunology
  • Radioimmunoassay*
  • Ultraviolet Rays

Substances

  • Antibodies, Antinuclear
  • Epitopes
  • Immune Sera
  • Polydeoxyribonucleotides
  • Pyrimidine Dimers
  • Poly dA-dT