Miner K M, Reading C L, Nicolson G L
Invasion Metastasis. 1981;1(3):158-74.
Mouse-mouse and rat-mouse hybridoma cell lines secreting complement-dependent cytotoxic or cell-binding monoclonal antibodies have been produced against cell surface components of two murine metastatic tumor systems: B16 melanoma and RAW117 lymphosarcoma. We have used both in vivo and in vitro spleen cell culture methods for immunization and have made a number of methodologic alterations to increase the yield and survival of hybridomas including media osmolality, pH, serum type, macrophage feeder layers and supplemented amino acids, vitamins and metabolites. Using complement-dependent cytotoxicity or lectin immobilization of tumor cells to polystyrene via a water-soluble carbodiimide for an enzyme-linked immunosorbant assay we were able to rapidly screen hybridoma cultures for specific antibody secretion.
已经制备出针对两种小鼠转移性肿瘤系统（B16黑色素瘤和RAW117淋巴肉瘤）细胞表面成分的、分泌补体依赖细胞毒性或细胞结合单克隆抗体的小鼠-小鼠和大鼠-小鼠杂交瘤细胞系。我们使用体内和体外脾细胞培养方法进行免疫，并对方法进行了一些改进，以提高杂交瘤的产量和存活率，包括培养基渗透压、pH值、血清类型、巨噬细胞饲养层以及补充氨基酸、维生素和代谢产物。通过水溶性碳二亚胺将肿瘤细胞固定在聚苯乙烯上进行酶联免疫吸附测定，利用补体依赖细胞毒性或凝集素固定化，我们能够快速筛选杂交瘤培养物中特异性抗体的分泌情况。
