Characterization of a monoclonal antibody to thymidine glycol monophosphate

Journal Article · Thu Nov 01 00:00:00 EST 1990 · Radiation Research; (USA)

DOI:https://doi.org/10.2307/3577856· OSTI ID:6320777

Chen, B X; Hubbard, K; Ide, H; Wallace, S S; Erlanger, B F ^[1]

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1. Columbia Univ. College of Physicians and Surgeons, New York, NY (USA)

A monoclonal antibody specific for thymine glycol (TG) in irradiated or OsO4-treated DNA was obtained by immunizing with thymidine glycol monophosphate (TMP-glycol) conjugated to bovine serum albumin by a carbodiimide procedure. Screening by dot-immunobinding and enzyme-linked immunosorbant assay (ELISA) procedures gave eight clones that bound OsO4- treated DNA. One of them, 2.6F.6B.6C, an IgG2a kappa, was characterized further. Hapten inhibition studies with OsO4-treated DNA showed that the antibody was specific for TMP-glycol. Among the various inhibitors tested, inhibition was in the order TMP-glycol greater than 5,6-dihydrothymidine phosphate greater than TMP greater than thymidine glycol greater than TG. Inhibition by 5,6-dihydrothymidine, thymidine, thymine, AMP, and CMP was negligible. In OsO4-treated DNA, as few as 0.5 TG per 10,000 bp were detectable by direct ELISA. Inhibition assays could detect as few as 1.5 TG per 10,000 bp. The antibody was equally reactive with native or denatured DNA containing TG. Among the X-irradiated homopolymers dC, dA, dG, and dT, only dT reacted with the antibody. Using an ELISA, the antibody could detect damage in irradiated DNA at the level of 20 Gy. Thus the antibody is of potential use in assays for DNA damage caused by X rays or other agents that damage DNA by free radical interactions.

• OSTI ID: 6320777
• Journal Information: Radiation Research; (USA), Vol. 124:2; ISSN 0033-7587
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
59 BASIC BIOLOGICAL SCIENCES
DNA
CHEMICAL RADIATION EFFECTS
MONOCLONAL ANTIBODIES
SPECIFICITY
THYMIDINE
ENZYME IMMUNOASSAY
ALBUMINS
INHIBITION
NUCLEOTIDES
OSMIUM OXIDES
RADICALS
X RADIATION
ANTIBODIES
AZINES
BIOASSAY
CHALCOGENIDES
CHEMISTRY
ELECTROMAGNETIC RADIATION
HETEROCYCLIC COMPOUNDS
IMMUNOASSAY
IONIZING RADIATIONS
NUCLEIC ACIDS
NUCLEOSIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OSMIUM COMPOUNDS
OXIDES
OXYGEN COMPOUNDS
PROTEINS
PYRIMIDINES
RADIATION CHEMISTRY
RADIATION EFFECTS
RADIATIONS
REFRACTORY METAL COMPOUNDS
RIBOSIDES
TRANSITION ELEMENT COMPOUNDS
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture
550200 - Biochemistry