Detection of oxidative DNA damage by a monoclonal antibody: role of lysyl residues in antigen binding.
Hydroxyl radical, a prominent entity of reactive oxygen species, is known to modify cellular DNA and has been implicated in several human diseases. A previously described monoclonal antibody (Mab) against reactive oxygen species-modified DNA (ROS-DNA), which preferentially recognizes ROS-modified epitopes on DNA, was used in this study. The epsilon-amino groups of lysine of the Mab were modified to study the role of these residues in Mab binding to ROS-DNA. The results demonstrate that modification of lysyl residues paralleled loss in Mab binding to ROS-DNA to the extent of 73%, suggesting the probable role of these positively charged amino acid residues in the complementarily determining regions of the Mab. The Mab was also used as an immunochemical probe to detect oxidative DNA damage in vivo in SLE. The Mab distinctly recognized five DNA samples out of eight from SLE patients and gave maximum inhibitions of 57, 58, 63, 64 and 70% in inhibition assay, while not reacting with DNA from normal, healthy population which served as negative control. High recognition of DNA isolates from SLE patients by the Mab having preferential binding to ROS-modified epitopes indicates increased oxidative stress in these patients leading to DNA damage which may contribute to the induction of antibodies cross-reacting with native DNA (nDNA).
Department of Biochemistry, Faculty of Medicine, Aligarh Muslim University, India.
J
J
Ahmad
B T
BT
Ashok
R
R
Ali
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology; D002417:Cattle; D004247:DNA / Q000276:immunology*; D004249:DNA Damage / Q000276:immunology*; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology; D008239:Lysine / Q000276:immunology*; D018384:Oxidative Stress
D000911:Antibodies, Monoclonal; D000941:Antigens; D004247:DNA; D008239:Lysine