Use of a monoclonal antibody to detect DNA damage caused by the anticancer drug cis‐diamminedichloroplatinum (II) in vivo and in vitro

FEBS Letters

Chao, Chuck C.-K.²  Shieh, Teh-Chien²  Huang, Haimei¹ 

[1] Institute of Radiation Biology, National Tsing Hua University, Hsinchu, Taiwan 30043, ROC;Tumor Biology Laboratory, Department of Biochemistry, Chang Gung Medical College, Taoyuan, Taiwan 33332, ROC

关键词: Cisplatin;    DNA repair;    Damage-recognition protein;    ELISA;    cisplatin;    cis-diamminedichloroplatinum (II);    CDRP;    cisplatin-damaged DNA recognition protein;    ELISA;    enzyme-linked immunoassay;    UV;    ultraviolet radiation;   

DOI  :  10.1016/0014-5793(94)01088-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown