Use of a monoclonal antibody to detect DNA damage caused by the anticancer drug cis-diamminedichloroplatinum (II) in vivo and in vitro
A monoclonal antibody, MAb62-5, was prepared and used to detect DNA damage due to the anticancer drug cis-diamminedichloroplatinum (II) (or cisplatin). ELISA competition indicated that the binding of MAb62-5 to cisplatin-DNA was competitively inhibited (50% control) by 210 nM of cisplatin bound to D...
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Published in | FEBS letters Vol. 354; no. 1; pp. 103 - 109 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
England
Elsevier B.V
31.10.1994
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Subjects | |
Online Access | Get full text |
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Summary: | A monoclonal antibody, MAb62-5, was prepared and used to detect DNA damage due to the anticancer drug
cis-diamminedichloroplatinum (II) (or cisplatin). ELISA competition indicated that the binding of MAb62-5 to cisplatin-DNA was competitively inhibited (50% control) by 210 nM of cisplatin bound to DNA, cisplatin/nucleotide (DIN) = 0.2. Using a DNA mobility shift assay, MAb62-5 binding activity was inhibited by 50% by ∼50-fold molar excess of cisplatin-DNA adducts (DIN = 0.08) whereas there was less than 5% inhibition by UV-DNA adducts or mock-treated DNA. In addition, MAb62-5 showed a similar affinity to the cisplatin-DNA adducts as compared to an endogenous cisplatin-damaged DNA recognition protein. Using ELISA with this antibody, we have demonstrated a 2-fold enhancement in excision repair of cisplatin-DNA adducts in resistant HeLa cells. This is supported by the measurement of repair-associated DNA strand breaks using alkaline elution and host cell reactivation of transfected plasmid DNA carrying cisplatin damage. These findings also provide a possible explanation for the complexicity of immunoassay in cells. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
ISSN: | 0014-5793 1873-3468 |
DOI: | 10.1016/0014-5793(94)01088-9 |