Monoclonal antibody to single-stranded DNA: a potential tool for DNA repair studies.
Growing evidence suggests that DNA repair capacity is an important factor in cancer risk and is therefore essential to assess. Immunochemical assays are amenable to the detection of repair products in complex matrices, such as urine, facilitating noninvasive measurements, although diet and extra-DNA sources of lesion can confound interpretation. The production of single-stranded, lesion-containing DNA oligomers characterises nucleotide excision repair (NER) and hence defines the repair pathway from which a lesion may be derived. Herein we describe the characterisation of a monoclonal antibody which recognises guanine moieties in single-stranded DNA. Application of this antibody in ELISA, demonstrated such oligomers in supernatants from repair-proficient cells post-insult. Testing of urine samples from volunteers demonstrated a relationship between oligomer levels and two urinary DNA damage products, thymine dimers and 8-oxo-2'-deoxyguanosine, supporting our hypothesis that NER gives rise to lesion-containing oligomers which are specific targets for the investigation of DNA repair.
Cooke
M S
MS
Oxidative Stress Group, Division of Chemical Pathology, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester, LE2 7LX, United Kingdom. msc5@le.ac.uk
Patel
K
K
Ahmad
J
J
Holloway
K
K
Evans
M D
MD
Lunec
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000378:metabolism*; D000918:Antibody Specificity; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000378:metabolism; D004249:DNA Damage / Q000276:immunology; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000276:immunology* / Q000378:metabolism* / Q000652:urine; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006151:Guanosine / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism; D006825:Hybridomas / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D016877:Oxidants / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D004277:DNA, Single-Stranded; D016877:Oxidants; D006151:Guanosine; C046215:8-hydroxyguanosine; D006861:Hydrogen Peroxide