DETECTION OF CYCLOBUTANE THYMINE DIMERS IN DNA OF HUMAN CELLS WITH MONOCLONAL ANTIBODIES RAISED AGAINST A THYMINE DIMER‐ CONTAINING TETRANUCLEOTIDE
DETECTION OF CYCLOBUTANE THYMINE DIMERS IN DNA OF HUMAN CELLS WITH MONOCLONAL ANTIBODIES RAISED AGAINST A THYMINE DIMER‐ CONTAINING TETRANUCLEOTIDE
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使用针对含胸腺嘧啶二聚体的四核苷酸产生的单克隆抗体检测人细胞 DNA 中的环丁烷胸腺嘧啶二聚体
DOI：
10.1111/j.1751-1097.1988.tb02873.x
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发表时间：
1988
影响因子：
3.3
通讯作者：
R. Baan
中科院分区：
文献类型：
--
作者：
L. Roza;Kees J. M. Wulp;S. J. MacFarlane;Paul H. M.Lohman;R. Baan
摘要：将小鼠浆细胞瘤细胞与用含有胸腺嘧啶二聚体的四核苷酸与载体蛋白偶联的小鼠的脾细胞融合后，分离出杂交细胞系（杂交瘤）。通过在竞争性酶联免疫吸附测定 (ELISA) 中测试各种抑制剂的效果来表征该杂交瘤产生的单克隆抗体。该抗体对单链 DNA 或聚 (dT) 中的胸腺嘧啶二聚体具有高度特异性，但不结合紫外线照射的 d(TpC)5。含有短胸腺嘧啶二聚体的序列观察到较少的结合。在光存在下用光活化酶或藤黄微球菌紫外线核酸内切酶对紫外线照射的 DNA 进行体外处理会导致抗原性消失。抗体与从紫外线照射的人成纤维细胞（254 nm）中分离出的 DNA 的结合与剂量呈线性关系。用抗体检测到，在辐射后孵育期间，这些细胞中胸腺嘧啶二聚体的去除最初很快，但速度迅速下降（10 J/m2 后 20 小时，残留二聚体约 50%）。免疫化学、ELISA 以及定量免疫荧光显微镜也证明了低剂量 UV-B 照射的人体皮肤中胸腺嘧啶二聚体的诱导。
Abstract— A hybrid cell line (hybridoma) has been isolated after fusion between mouse‐plasmacytoma cells and spleen cells from mice immunized with a thymine dimer‐containing tetranucleotide coupled to a carrier protein. Monoclonal antibodies produced by this hybridoma were characterized by testing the effect of various inhibitors in a competitive enzyme‐linked immunosorbent assay (ELISA). The antibodies have a high specificity for thymine dimers in single‐stranded DNA or poly(dT), but do not bind UV‐irradiated d(TpC)5. Less binding is observed with short thymine dimer‐containing sequences. In vitro treatment of UV‐irradiated DNA with photoreactivating enzyme in the presence of light, or with Micrococcus luteus UV‐endonuclease results in disappearance of antigenicity. Antibody‐binding to DNA isolated from UV‐irradiated human fibroblasts (at 254 nm) is linear with dose. Removal of thymine dimers in these cells during a post‐irradiation incubation, as detected with the antibodies, is fast initially but the rate rapidly decreases (about 50% residual dimers at 20 h after 10 J/m2). The induction of thymine dimers in human skin irradiated with low doses of UV‐B, too, was demonstrated immunochemically, by ELISA as well as by quantitative immunofluorescence microscopy.
开发用于检测哺乳动物细胞 DNA 中光产物的放射免疫测定法。
DOI：
10.1016/0005-2787(81)90066-6
发表时间：
1981
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Mitchell,DL;Clarkson,JM
通讯作者：
Clarkson,JM