Title
Quantitative detection of DNA damage in cells after exposure to ionizing radiation by means of an improved immunochemical assay
Author
Medisch Biologisch Laboratorium TNO
van Loon, A.A.W.M.
Groenendijk, R.H.
Timmerman, A.J.
van der Schans, G.P.
Lohman, P.H.M.
Baan, R.A.
Publication year
1992
Abstract
A simple, sensitive and fast immunochemical method has been developed to quantify the amount of DNA damage in cells of human blood after in vitro exposure to ionizing radiation. The technique is based on the enhancement of the radiation-induced single-strandedness, which occurs in DNA regions flanking strand breaks, by a controlled further unwinding of the DNA in an alkaline solution. Subsequently, the DNA is attached to the wall of polystyrene cups by passive adsorption. DNA damage is then quantified by determining the extent of single-strandedness with a mononuclear antibody, D1B, directed against single-stranded DNA. D1B binding is assayed with a 'second' antibody, labelled with either an enzyme or europium. The latter gives slightly more reproducible results. No radioactive labelling of DNA is required and the assay takes only 3.5 h after the collection of blood. Damage can be detected after doses as low as 0.5 Gy. The potential broader application of the method is discussed. Chemicals/CAS: Antibodies, Monoclonal; DNA, 9007-49-2; DNA, Single-Stranded; Europium, 7440-53-1.
Subject
Anti single-stranded DNA antibody
ELISA
EuLISA
Ionising radiation
Single-stranded DNA
DNA damage
DNA structure
Human
Human cell
Immunochemistry
Ionizing radiation
Lymphocyte
Priority journal
Technique
Antibodies, Monoclonal
DNA
DNA Damage
DNA, Single-Stranded
Enzyme-Linked Immunosorbent Assay
Europium
Human
Immunoassay
Leukocytes
Support, Non-U.S. Gov't
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DOI
https://doi.org/10.1016/0921-8777(92)90040-a
TNO identifier
231879
ISSN
0921-8777
Source
Mutation Research - DNA Repair, 274 (274), 19-27
Document type
article