In vivo and in vitro production and detection of monoclonal antibodies to surface components on metastatic variants of murine tumor cells.

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.