Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody

Journal Article · Mon Jun 01 00:00:00 EDT 1987 · Experimental Cell Research; (USA)

DOI:https://doi.org/10.1016/0014-4827(87)90314-4· OSTI ID:6519784

Frankfurt, O S ^[1]

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1. Roswell Park Memorial Institute, Buffalo, NY (USA)

A new method for the measurement of DNA damage in individual cells treated with alkylating agents is described. The method is based on the binding of anti-DNA monoclonal antibody to DNA in situ. Binding of antibody was evaluated by flow cytometry with indirect immunofluorescence. No binding of antibody to DNA in non-treated HeLa S3 cells was detected. Treatment of cells with HN2 or L-phenylalanine mustard induced binding of antibody to DNA in situ. Binding of antibody was observed after treating cells with doses of drugs which reduced the surviving fraction below 20%. Intensity of binding increased in proportion to the drug dose. In HN2-treated cells a cell subset with the lowest antibody binding was observed among cells in G1 phase. Binding of antibody to DNA in HN2-treated cells was eliminated by single-strand (ss) specific S1 nuclease. In competition assay, antibody was inhibited by thermally denatured DNA, but not by native double-stranded (ds) DNA, RNA, nucleosides and deoxyribohomopolymers. Immunoreactivity of cells with the monoclonal antibody F7-26 may be a useful probe for the assessment of cell damage induced by alkylating agents, especially in heterogeneous cell populations.

• OSTI ID: 6519784
• Journal Information: Experimental Cell Research; (USA), Vol. 170:2; ISSN 0014-4827
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKYLATING AGENTS
BIOLOGICAL EFFECTS
DNA
ENZYME IMMUNOASSAY
CELL FLOW SYSTEMS
FLUORESCENCE SPECTROSCOPY
HELA CELLS
HYBRIDOMAS
IMMUNOLOGY
MONOCLONAL ANTIBODIES
ANIMAL CELLS
ANTIBODIES
BIOASSAY
EMISSION SPECTROSCOPY
IMMUNOASSAY
NUCLEIC ACIDS
ORGANIC COMPOUNDS
SPECTROSCOPY
560300* - Chemicals Metabolism & Toxicology