Polyclonal Antibodies to Adenine N1-Oxide: Characterization and Use for the Measurement of DNA Damage

Autor:	Didier Molko, Nathalie Signorini, Jean Cadet
Rok vydání:	1998
Předmět:	chemistry.chemical_classification Antiserum biology Chemistry DNA damage General Medicine Toxicology Adenosine Molecular biology chemistry.chemical_compound Biochemistry Antigen Polyclonal antibodies medicine biology.protein Nucleotide Hapten DNA medicine.drug
Zdroj:	Chemical Research in Toxicology. 11:1169-1175
ISSN:	1520-5010 0893-228X
DOI:	10.1021/tx980044+
Popis:	Adenine N1-oxide is a DNA lesion whose formation involves the specific oxidation of the adenine base by hydrogen peroxide under nonradical conditions. The damage may be measured using a HPLC/32P-postlabeling method, which however cannot be used for routine analysis. We propose herein as an alternative an immunological assay which allows a rapid evaluation of the level of adenine N1-oxide in DNA exposed to oxidative stress. Two polyclonal antibodies were raised using two different strategies for the coupling of the hapten to the protein. The first approach is based on the universal method of Erlanger and Beiser, whereas the preparation of the second antigen involves the conjugation of a morpholino derivative of adenosine N1-oxide to the carrier protein. The affinity and the specificity of those antibodies were determined by competitive enzyme-linked immunosorbent assay. The antibody obtained by the traditional method shows some cross-reactivity with normal nucleotides, whereas for the other antiserum, the ...
Databáze:	OpenAIRE
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