Lane R D, Crissman R S, Lachman M F
J Immunol Methods. 1984 Aug 3;72(1):71-6. doi: 10.1016/0022-1759(84)90434-4.
In order to improve the yield of hybridomas for monoclonal antibody production, 8 different sources and molecular weights of polyethylene glycol (PEG) were compared as fusing agents. Sp2/0 myeloma cells were fused with murine splenic lymphocytes immunized with sheep red blood cells. The Kodak 1450 PEG produced the maximum number of hybridomas. The optimal technique consisted of slowly adding 1 ml of freshly prepared fusogen (5 g Kodak 1450 PEG, 0.5 ml dimethylsulfoxide, and 5 ml of phosphate-buffered saline, pH 7.0) to the cells over a 1 min period, incubating the mixture at 37 degrees C for 90 s, then gradually diluting the mixture in 50 ml of Hanks' buffered salt solution. After 10 min, the cells are centrifuged, resuspended in selective medium with feeder macrophages and cultured. This procedure routinely produces between 600-3,000 hybridomas per fusion.
为提高用于单克隆抗体制备的杂交瘤产量，比较了8种不同来源和分子量的聚乙二醇（PEG）作为融合剂的效果。将Sp2/0骨髓瘤细胞与用绵羊红细胞免疫的小鼠脾淋巴细胞进行融合。柯达1450 PEG产生的杂交瘤数量最多。最佳技术包括在1分钟内将1毫升新鲜制备的融合剂（5克柯达1450 PEG、0.5毫升二甲基亚砜和5毫升pH 7.0的磷酸盐缓冲盐水）缓慢加入细胞中，将混合物在37℃孵育90秒，然后在50毫升汉克斯缓冲盐溶液中逐渐稀释混合物。10分钟后，将细胞离心，重悬于含有饲养巨噬细胞的选择培养基中并进行培养。该程序每次融合常规可产生600 - 3000个杂交瘤。
