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

Abstract

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.

Authors:
 [1]
  1. Roswell Park Memorial Institute, Buffalo, NY (USA)
Publication Date:
OSTI Identifier:
6519784
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research; (USA)
Additional Journal Information:
Journal Volume: 170:2; Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Frankfurt, O S. Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody. United States: N. p., 1987. Web. doi:10.1016/0014-4827(87)90314-4.
Frankfurt, O S. Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody. United States. doi:10.1016/0014-4827(87)90314-4.
Frankfurt, O S. Mon . "Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody". United States. doi:10.1016/0014-4827(87)90314-4.
@article{osti_6519784,
title = {Detection of DNA damage in individual cells by flow cytometric analysis using anti-DNA monoclonal antibody},
author = {Frankfurt, O S},
abstractNote = {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.},
doi = {10.1016/0014-4827(87)90314-4},
journal = {Experimental Cell Research; (USA)},
issn = {0014-4827},
number = ,
volume = 170:2,
place = {United States},
year = {1987},
month = {6}
}