An immunochemical approach to the study of DNA damage and repair

Technical Report · Sun Jan 01 00:00:00 EST 1989

DOI:https://doi.org/10.2172/5359966· OSTI ID:5359966

Wallace, S. S.; Erlanger, B. F. ^[1]

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1. Vermont Univ., Burlington, VT (USA). Dept. of Microbiology and Molecular Genetics

The most studied radiation-induced modified DNA base is thymine glycol. Thymine glycols are produced in relatively high yields in irradiated DNA and are also formed as a consequence of oxidative stress. Thymine glycol has been shown to be an in vitro replicative block to DNA polymerases as well as a cytotoxic lesion. DNA glycosylases that remove thymine glycol from damaged DNA are found in both prokaryotes an deukaryotes. Thus this lesion provides a good model for studying the potential biological consequences of pyrimidine ring saturation products. In order to elicit antibodies that would react with unique modified base on a damaged DNA molecule, we have chosen to chemically synthesize the hapten of interest and conjugate it to a protein carrier. Thymine glycol monophosphate was synthesized, conjugated by the carbodiimide method to bovine serum albumin (BSA), and used as an immunogen. Initially, polyclonal antibodies were produced in rabbits. These antibodies had high affinity and specificity as measured by both albumin (RSA) and by enzyme immunoassay using either the conjugate or DNA oxidized inhibited by thymine glycol, thymidine glycol and thymine glycol monophosphate determinants for the phosphate group in addition to the thymidine glycol. In both the direct and competitive assays, this antibody reacts with osmium tetroxide-treated DNA containing cis-thymine glycols and DNA X-irradiated in vitro at a femtomole level of sensitivity. 24 refs.

• Research Organization: Vermont Univ., Burlington, VT (USA). Dept. of Microbiology and Molecular Genetics; Columbia Univ., New York, NY (USA). Dept. of Microbiology
• Sponsoring Organization: DOE/ER
• DOE Contract Number: FG02-88ER60742
• OSTI ID: 5359966
• Report Number(s): DOE/ER/60742-2; ON: DE90003347; TRN: 90-000921
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CHROMOSOME BREAKAGE
IMMUNOASSAY
RADIOPHARMACEUTICALS
PRODUCTION
DNA REPAIR
GLYCOLS
MONOCLONAL ANTIBODIES
NUCLEOTIDES
PROGRESS REPORT
QUANTITATIVE CHEMICAL ANALYSIS
RADIOLYSIS
ALCOHOLS
ANTIBODIES
BIOASSAY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CHEMICAL ANALYSIS
CHEMICAL RADIATION EFFECTS
CHEMICAL REACTIONS
CHEMISTRY
CHROMOSOMAL ABERRATIONS
DECOMPOSITION
DOCUMENT TYPES
DRUGS
HYDROXY COMPOUNDS
LABELLED COMPOUNDS
MUTATIONS
ORGANIC COMPOUNDS
RADIATION CHEMISTRY
RADIATION EFFECTS
RECOVERY
REPAIR
550200* - Biochemistry
550201 - Biochemistry- Tracer Techniques