Establishment of a monoclonal antibody recognizing ultraviolet light-induced (6-4) photoproducts

Mutation Research/DNA Repair Reports Pub Date : 1988-11-01 DOI:10.1016/0167-8817(88)90028-4

Toshio Mori, Tsukasa Matsunaga, Tohru Hirose, Osamu Nikaido

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引用次数: 45

Abstract

We obtained a monoclonal antibody directed against UV-induced DNA damage. Analysis of the antigenic determinant in UV-irradiated DNA recognized by this antibody, 64M-1, revealed that it bound UV-irradiated oligo- or poly-nucleotides containing thymine—thymine or thymine—cytosine sequences. The antibody failed to bind DNA irradiated with 313 nm UV in the presence of acetophenone, which contained predominantly thymine dimers as DNA damage. The binding activity of this antibody to 254-nm UV-irradiated DNA decreased with 313-nm UV irradiation, and the decrease of this binding activity correlated with the decrease of fluorescence corresponding to (6-4) photoproducts. These results suggest that the antigenic determinant recognized by this monoclonal antibody is a (6-4) photoproduct. Using autoradiography with ³H-antibody, we could detect the formation of the (6-4) photoproduct in individual human cells irradiated with 254-nm UV doses as low as 20 J/m².

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一种识别紫外光诱导(6-4)光产物单克隆抗体的建立

我们获得了一种针对紫外线诱导的DNA损伤的单克隆抗体。对该抗体识别的紫外线照射DNA的抗原决定因子64M-1进行分析，发现它结合了含有胸腺嘧啶-胸腺嘧啶或胸腺嘧啶-胞嘧啶序列的紫外线照射寡核苷酸或多核苷酸。在含有胸腺嘧啶二聚体的苯乙酮的313 nm紫外照射下，抗体不能与DNA结合。该抗体对254 nm紫外光照射DNA的结合活性在313 nm紫外光照射下降低，其结合活性的降低与(6-4)光产物对应的荧光降低有关。这些结果表明，该单克隆抗体识别的抗原决定因子是一个(6-4)光产物。利用3h抗体放射自显影技术，我们可以检测到在低至20 J/m2的254 nm紫外线照射下个体人体细胞中(6-4)光产物的形成。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Mutation Research/DNA Repair Reports

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