Olsson L, Kaplan H S
Proc Natl Acad Sci U S A. 1980 Sep;77(9):5429-31. doi: 10.1073/pnas.77.9.5429.
We report the establishment of human-human hybridomas producing monoclonal antibody of predefined antigenic specificity. The U-266 human myeloma cell line was incubated in the presence of 8-azaguanine, and a rapidly growing, 8-azaguanine-resistant, hypoxanthine/amethopterin/thymidine (HAT) medium-sensitive mutant line, U-266AR1, was selected. These cells were fused with lymphoid cells from uninvolved spleens removed at staging laparotomy from patients with untreated Hodgkin's disease who had been previously sensitized to the chemical allergen 2,4-dinitrochlorobenzne. Hybrid cell cultures growing in HAT medium were screened for IgG production. Positive cultures were selected and their supernatants were tested in a solid-phase radioimmunoassay for reactivity with dinitrophenyl hapten coupled to bovine serum albumin. Cultures producing specific antibody were subcloned and expanded, and their antibody products were shown to be monoclonal by biosynthetic labeling and sodium dodecyl sulfate/polyacrylamide gel electrophoresis.
我们报道了产生具有预定义抗原特异性单克隆抗体的人-人杂交瘤的建立。将U-266人骨髓瘤细胞系在8-氮杂鸟嘌呤存在下培养，并筛选出一种生长迅速、对8-氮杂鸟嘌呤耐药、对次黄嘌呤/氨甲蝶呤/胸腺嘧啶核苷（HAT）培养基敏感的突变株系U-266AR1。这些细胞与来自未经治疗的霍奇金病患者分期剖腹探查时切除的未受累脾脏的淋巴细胞融合，这些患者先前已对化学过敏原2,4-二硝基氯苯致敏。在HAT培养基中生长的杂交细胞培养物进行IgG产生筛选。选择阳性培养物，其上清液在固相放射免疫测定中检测与偶联至牛血清白蛋白的二硝基苯基半抗原的反应性。产生特异性抗体的培养物进行亚克隆和扩增，其抗体产物通过生物合成标记和十二烷基硫酸钠/聚丙烯酰胺凝胶电泳显示为单克隆。
