Generation of monoclonal antibodies reacting with normal and cancer cells of human breast

Abstract

From the fusion of the murine myeloma P3- X63-Ag8-U1 with spleen cells from a mouse immunized with the human breast cancer cell line MCF-7, 88 hybridomas producing antibodies reacting with the immunizing cells were obtained. After a first screening on human leukocytes, red blood cells, and platelets and on human and fetal calf serum, three monoclonal antibodies, MBr1, MBr2, and MBr3, with specificity for the immunizing cells were isolated and further characterized. The three monoclonals were tested by isotopic antiglobulin assay and immunofluorescence on a panel of normal cells or cell membrane preparations, including milk epithelial and foam cells; on plasma and milk proteins; on cells or cell membrane preparations from fresh surgical specimens of breast, kidney, and ovarian carcinomas; and on various tumor cell lines. MBr1 and MBr2 had a superimposable reactivity and showed specificity for a structure which seems to characterize both normal and neoplastic mammary gland epithelial cells.