Immunochemical detection of glyoxal DNA damage

Nalini Mistry, Mark D. Evans, Helen R. Griffiths, H. Kasai, Karl E. Herbert, Joseph Lunec

Research output: Contribution to journalArticlepeer-review


The relevance of reactive oxygen species (ROS) in the pathogenesis of inflammatory diseases is widely documented. Immunochemical detection of ROS DNA adducts has been developed, however, recognition of glyoxal-DNA adducts has not previously been described. We have generated a polyclonal antibody that has shown increased antibody binding to ROS-modified DNA in comparison to native DNA. In addition, dose-dependent antibody binding to DNA modified with ascorbate alone was shown, with significant inhibition by desferrioxamine, catalase, and ethanol. Minimal inhibition was observed with uric acid, 1,10-phenanthroline and DMSO. However, antibody binding in the presence of EDTA increased 3500-fold. The involvement of hydrogen peroxide and hydroxyl radical in ascorbate-mediated DNA damage is consistent with ascorbate acting as a reducing agent for DNA-bound metal ions. Glyoxal is known to be formed during oxidation of ascorbate. Glyoxylated DNA, that previously had been proposed as a marker of oxidative damage, was recognised in a dose dependent manner using the antibody. We describe the potential use of our anti-ROS DNA antibody, that detects predominantly Fenton-type mediated damage to DNA and report on its specificity for the recognition of glyoxal-DNA adducts.
Original languageEnglish
Pages (from-to)1267-1273
Number of pages7
JournalFree Radical Biology and Medicine
Issue number9-10
Publication statusPublished - May 1999


  • DNA
  • glyoxal
  • antibodies
  • free radicals
  • immunochemical detection
  • ascorbic acid
  • fenton chemistry


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