Groopman J D, Haugen A, Goodrich G R, Wogan G N, Harris C C
Cancer Res. 1982 Aug;42(8):3120-4.
Five monoclonal antibodies were obtained after fusion of mouse P3 x 63 myeloma cells with spleen cells isolated from BALB/c mice that had been immunized with aflatoxin B1-adducted DNA complexes with methylated bovine serum albumin. Selected hybridomas were found to produce monoclonal antibodies specific for modified DNA containing both the 2,3-dihydro-2-(N7-guanyl)-3-hydroxyaflatoxin B1 and the putative 2,3-dihydro-2-(N5-formyl-2',5',6'-triamino-4'-oxo-N5-pyrimidyl)-3-hydroxyaflatoxin B1, suggesting that these DNA adducts share a common antigenic determinant. Enzyme immunoassay conditions using these monoclonal antibodies were optimized, and DNA isolated from the livers of rats given dosages of aflatoxin B1 ranging from 0.01 to 1.0 mg aflatoxin B1 per kg body weight was tested. A level of modification in DNA of 1 aflatoxin B1 residue per 1,355,000 nucleotides can be quantitatively measured. Monoclonal antibodies will be useful probes for studying the molecular interactions of aflatoxin B1 with DNA and the occurrence of aflatoxin B1:DNA adducts in tissues and cells of humans exposed to this environmental carcinogen.
将小鼠P3 x 63骨髓瘤细胞与从用黄曲霉毒素B1加合物与甲基化牛血清白蛋白免疫的BALB/c小鼠中分离的脾细胞融合后，获得了五种单克隆抗体。发现所选杂交瘤产生对含有2,3-二氢-2-(N7-鸟嘌呤基)-3-羟基黄曲霉毒素B1和推定的2,3-二氢-2-(N5-甲酰基-2',5',6'-三氨基-4'-氧代-N5-嘧啶基)-3-羟基黄曲霉毒素B1的修饰DNA具有特异性的单克隆抗体，这表明这些DNA加合物具有共同的抗原决定簇。使用这些单克隆抗体的酶免疫测定条件得到了优化，并对给予每千克体重0.01至1.0毫克黄曲霉毒素B1剂量的大鼠肝脏中分离的DNA进行了检测。每1,355,000个核苷酸中1个黄曲霉毒素B1残基的DNA修饰水平可以进行定量测量。单克隆抗体将成为研究黄曲霉毒素B1与DNA的分子相互作用以及接触这种环境致癌物的人类组织和细胞中黄曲霉毒素B1:DNA加合物发生情况的有用探针。
