Bepaling van DNA-Schade in aan Ioniserende Straling Blootgestel - de Cellen met Behulp van Monoklonale Antilichamen Gericht Tegen Enkelstrengig DNA (Detection of DNA Damage in Cells Exposed to Ionizing Radiation by Use of Anti-Single-Stranded-DNA Monoclonal Antibody)

摘要

An immunochemical method has been developed for quantitative detection of DNA damage in mammalian cells. The method is based on the binding of a monoclonal antibody to single-stranded DNA. The clone producing this antibody, D1B, was obtained as a byproduct from fusion of mouse myeloma cells with spleen cells isolated from a mouse immunized with chemically modified DNA. The technique is based upon the determination of the percentage single-strandedness resulting from the partial unwinding of cellular DNA under alkaline conditions, a time-dependent process. Single-strand and double-strand DNA breaks, or lesions converted into such breaks in alkaline medium, form initiation points for the unwinding. The extent of the unwinding under controlled conditions is a measure, therefore, of the amount of such sites. The method is rapid, does not require radioactive labelling of DNA or physical separation of single- from double-stranded molecules, is sufficiently sensitive to detect damage induced by 1 Gy of ionizing radiation and needs only small amount of cells.