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Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer-containing tetranucleotide

Journal Article:

Abstract

A hybrid cell line (hybridoma) has been isolated after fusion between mouse-plasmacytoma cells and spleen cells from mice immunized with a thymine dimer-containing tetranucleotide coupled to a carrier protein. Monoclonal antibodies produced by this hybridoma were characterized by testing the effect of various inhibitors in a competitive enzyme-linked immunosorbent assay (ELISA). The antibodies have a high specificity for thymine dimers in single-stranded DNA or poly(dT), but do not bind UV-irradiated d(TpC)/sub 5/. Less binding is observed with short thymine dimer-containing sequences. In vitro treatment of UV-irradiated DNA with photoreactivating enzyme in the presence of light, or with Micrococcus luteus UV-endonuclease results in disappearance of antigenicity. Antibody-binding to DNA isolated from UV-irradiated human fibroblasts (at 254 nm) is linear with dose. Removal of thymine dimers in these cells during a post-irradiation incubation, as detected with the antibodies, is fast initially but the rate rapidly decreases (about 50% residual dimers at 20 h after 10 J/m/sup 2/). The induction of thymine dimers in human skin irradiated with low doses of UV-B, too, was demonstrated immunochemically, by ELISA as well as by quantitative immunofluorescence microscopy.
Publication Date:
Nov 01, 1988
Product Type:
Journal Article
Reference Number:
AIX-20-052091; EDB-89-094729
Resource Relation:
Journal Name: Photochem. Photobiol.; (United Kingdom); Journal Volume: 48:5
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; DIMERS; FIBROBLASTS; MONOCLONAL ANTIBODIES; NEAR ULTRAVIOLET RADIATION; CELL CULTURES; CYCLOALKANES; DNA; HYBRIDOMAS; MAN; SKIN; THYMINE; ALKANES; ANIMAL CELLS; ANIMALS; ANTIBODIES; AZINES; BODY; CONNECTIVE TISSUE CELLS; ELECTROMAGNETIC RADIATION; HETEROCYCLIC COMPOUNDS; HYDROCARBONS; HYDROXY COMPOUNDS; MAMMALS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANS; PRIMATES; PYRIMIDINES; RADIATIONS; SOMATIC CELLS; ULTRAVIOLET RADIATION; URACILS; VERTEBRATES; 560120* - Radiation Effects on Biochemicals, Cells, & Tissue Culture
OSTI ID:
6253390
Research Organizations:
TNO Medical Biological Lab., Rijswijk (Netherlands); Erasmus Univ., Rotterdam (Netherlands)
Country of Origin:
United Kingdom
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: PHCBA
Submitting Site:
GBN
Size:
Pages: 627-633
Announcement Date:

Journal Article:

Citation Formats

Roza, L, Wulp, K.J.M. van der, MacFarlane, S J, Lohman, P H.M., and Baan, R A. Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer-containing tetranucleotide. United Kingdom: N. p., 1988. Web.
Roza, L, Wulp, K.J.M. van der, MacFarlane, S J, Lohman, P H.M., & Baan, R A. Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer-containing tetranucleotide. United Kingdom.
Roza, L, Wulp, K.J.M. van der, MacFarlane, S J, Lohman, P H.M., and Baan, R A. 1988. "Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer-containing tetranucleotide." United Kingdom.
@misc{etde_6253390,
title = {Detection of cyclobutane thymine dimers in DNA of human cells with monoclonal antibodies raised against a thymine dimer-containing tetranucleotide}
author = {Roza, L, Wulp, K.J.M. van der, MacFarlane, S J, Lohman, P H.M., and Baan, R A}
abstractNote = {A hybrid cell line (hybridoma) has been isolated after fusion between mouse-plasmacytoma cells and spleen cells from mice immunized with a thymine dimer-containing tetranucleotide coupled to a carrier protein. Monoclonal antibodies produced by this hybridoma were characterized by testing the effect of various inhibitors in a competitive enzyme-linked immunosorbent assay (ELISA). The antibodies have a high specificity for thymine dimers in single-stranded DNA or poly(dT), but do not bind UV-irradiated d(TpC)/sub 5/. Less binding is observed with short thymine dimer-containing sequences. In vitro treatment of UV-irradiated DNA with photoreactivating enzyme in the presence of light, or with Micrococcus luteus UV-endonuclease results in disappearance of antigenicity. Antibody-binding to DNA isolated from UV-irradiated human fibroblasts (at 254 nm) is linear with dose. Removal of thymine dimers in these cells during a post-irradiation incubation, as detected with the antibodies, is fast initially but the rate rapidly decreases (about 50% residual dimers at 20 h after 10 J/m/sup 2/). The induction of thymine dimers in human skin irradiated with low doses of UV-B, too, was demonstrated immunochemically, by ELISA as well as by quantitative immunofluorescence microscopy.}
journal = {Photochem. Photobiol.; (United Kingdom)}
volume = {48:5}
journal type = {AC}
place = {United Kingdom}
year = {1988}
month = {Nov}
}