Strand M
Proc Natl Acad Sci U S A. 1980 Jun;77(6):3234-8. doi: 10.1073/pnas.77.6.3234.
Tumor-cell proteins that were antigenic in a syngeneic animal were identified by immunoprecipitation with monoclonal antibodies. Spleen cells of BALB/c mice immunized with plasma membranes of Kirsten RNA sarcoma virus-transformed BALB/3T3 cells were fused with NS-1 myeloma cells. Antibodies secreted into the culture fluid from these hybridomas were distinguished by their reactivity against proteins of different target cells. A total of 191 cultures were established; 143 produced antibodies that bound to BALB/3T3 cells transformed by the RNA sarcoma virus, of which antibodies from 82 bound to BALB/3T3 transformed with simian virus 40, and antibodies from 56 bound to BALB/3T3 cells. Thus, more than 50% of the cultures produced antibodies that possibly were specific to antigens of the transformed cell. Twenty different hybridomas have been cloned, and antibodies, from eight of these were found to immunoprecipitate five different proteins. A protein of approximately 32,000 daltons was precipitated from BALB/3T3 cells transformed by the RNA sarcoma virus, simian virus 40, or methylcholanthrene but not from untransformed BALB/3T3 cells. A protein of about 300,000 daltons was precipitated from all four cell lines; precipitation was enhanced in the viral transformed cells. Proteins of approximately 57,000, 54,000, and 8500 daltons were immunoprecipitated from all four cell lines.
通过用单克隆抗体进行免疫沉淀，鉴定出在同基因动物中具有抗原性的肿瘤细胞蛋白。用 Kirsten RNA 肉瘤病毒转化的 BALB/3T3 细胞的质膜免疫的 BALB/c 小鼠的脾细胞与 NS-1 骨髓瘤细胞融合。这些杂交瘤分泌到培养液中的抗体通过它们对不同靶细胞蛋白质的反应性来区分。总共建立了 191 个培养物；143 个产生了与 RNA 肉瘤病毒转化的 BALB/3T3 细胞结合的抗体，其中 82 个抗体的抗体与猿猴病毒 40 转化的 BALB/3T3 细胞结合，56 个抗体的抗体与 BALB/3T3 细胞结合。因此，超过 50%的培养物产生了可能对转化细胞抗原具有特异性的抗体。已经克隆了 20 种不同的杂交瘤，并且发现其中 8 种杂交瘤的抗体能够免疫沉淀 5 种不同的蛋白质。一种约 32,000 道尔顿的蛋白质从 RNA 肉瘤病毒、猿猴病毒 40 或甲基胆蒽转化的 BALB/3T3 细胞中沉淀出来，但未从未转化的 BALB/3T3 细胞中沉淀出来。一种约 300,000 道尔顿的蛋白质从所有四种细胞系中沉淀出来；在病毒转化的细胞中沉淀增强。约 57,000、54,000 和 8500 道尔顿的蛋白质从所有四种细胞系中被免疫沉淀出来。
