Comparison of various immunochemical assays for the detection of ethylene oxide-DNA adducts with monoclonal antibodies against imidazole ring-opened N7-(2-hydroxyethyl)guanosine : application in a biological monitoring study

article




Delft, J.H.M. van

Winden, M.J.M. van

Luiten-Schuite, A.

Ribeiro, L.R.

Baan, R.A.
Monoclonal antibodies have been developed for the analysis of the predominant lesion in DNA induced by ethylene oxide (EtOx), namely N7-(2-hydroxyethyl)guanine (N7-EtOHGua). Two monoclonal antibodies raised against imidazole ring-opened N7-(2-hydroxyethyl)guanine (RON7-EtOHGua), N7EO-E and N7EO-11, and the previously isolated antibody N7E-102 were characterized by competitive ELISA with various inhibitors. N7EO-E and N7EO-11 recognize 2-hydroxyethyl lesions better than ethyl or methyl lesions, while N7E-102 recognizes 2-hydroxyethyl and ethyl modifications equally well. All antibodies show a preference for imidazole ring-opened adducts, bind better to adducts in DNA compared to alkylated nucleosides or bases and bind 106- to 3 x 106-fold less well to unmodified DNA. The sensitivity of detection of RON7-EtOHGua in DNA and in the nuclei of cells in situ by antibody N7EO-E was investigated in several assays. The immunoslot blot assay was the most sensitive method (0.34 RON7-EtOHGua per 106 nucleotides was detectable), followed by competitive ELISA, direct ELISA and in situ detection by immunofluorescence microscopy. The immunoslot blot assay was used to analyse N7-EtOHGua levels in white blood cell DNA from individuals exposed to EtOx (2-5 p.p.m.) and of controls. This exposure did not result in a statistically significant increase in the N7-EtOHGua level.
Chemicals/CAS: 7 (2 hydroxyethyl)guanine, 53498-52-5; ethylene oxide, 16002-48-5, 32587-16-9, 36376-18-8, 75-21-8; Antibodies, Monoclonal; DNA Adducts; DNA, 9007-49-2; Ethylene Oxide, 75-21-8; Guanine, 73-40-5; N(7)-hydroxyethylguanine, 53498-52-5
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AnimalAntibodies, MonoclonalAntibody SpecificityCell NucleusComparative StudyDNADNA AdductsDNA DamageEnzyme-Linked Immunosorbent AssayEthylene OxideGuanineHumanImmunohistochemistryLeukocytesMaleMiceMice, Inbred StrainsMicroscopy, FluorescenceOccupational ExposureSensitivity and Specificity
TNO Identifier
36167
Repository link
https://resolver.tno.nl/uuid:832c6680-75d5-4302-841c-be41ef737498
ISSN
01433334
Source
Carcinogenesis, 15(9), pp. 1867-1873.
Pages
1867-1873
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