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Quantitative Measurement of Ultraviolet‐Induced Damage in Cellular DNA by an Enzyme Immunodot Assay

A simple enzyme immunoassay procedure was developed for the quantitative determination of 254‐nm uv‐induced DNA damage in cells. With the use of specific antibodies to uv‐irradiated DNA and horseradish peroxidase‐conjugated antibody to rabbit IgG, the extent of damaged DNA in uv‐irradiated rat spleen mononuclear cells was quantitatively measurable. Through the use of this method, the amount of damaged DNA present in 2 × 10(5) cells irradiated at a dose of 75 J/m2 was estimated to be 7 ng equivalents of the standard uv‐irradiated DNA. In addition, when the cells, irradiated at 750 J/m2, were incubated for 1 h, the antigenic activity of DNA decreased by 40%, suggesting that a repair of the damaged sites in DNA had proceeded to some extent in the cells.