Immunochemical quantitation of thymine glycol in oxidized and X-irradiated DNA.

Abstract:

:The present study demonstrates the usefulness of immunochemical assays for quantitating modified bases in oxidized and X-irradiated DNA. Escherichia coli, phi X174 RF I, PM2, and M13 DNA containing thymine glycols introduced by OsO4 oxidation were used as antigens in a direct enzyme-linked immunosorbent assay (ELISA). The number of thymine glycols per DNA molecule was determined by reactivity with antithymine glycol antibody standardized either to the acetol fragment assay or to the number of Escherichia coli endonuclease III-sensitive sites. The number of thymine glycols was also determined in phi X174 RF I DNA X-irradiated in either phosphate or Tris buffer under air. Using a direct ELISA with phi X174 RF I DNA irradiated in a phosphate buffer solution, the anti-thymine glycol antibody detected damage at the level of 40 Gy. The immunochemical assay was sensitive, specific, quantitative, and independent of DNA structure.

journal_name

Radiat Res

journal_title

Radiation research

authors

Hubbard K,Huang H,Laspia MF,Ide H,Erlanger BF,Wallace SS

subject

Has Abstract

pub_date

1989-05-01 00:00:00

pages

257-68

issue

2

eissn

0033-7587

issn

1938-5404

journal_volume

118

pub_type

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