GENERATION, CHARACTERIZATION, AND UTILIZATION OF MONOCLONAL ANTIBODIES REACTIVE WITH HUMAN MAMMARY CARCINOMA ASSOCIATED ANTIGENS

Abstract

Publisher Summary This chapter describes an experiment in which splenic lymphocytes of mice, immunized with membrane-enriched fractions of human metastatic mammary carcinoma cells, were fused with murine non- immunog1obulin secretor myeloma cells. Following initial screening and double cloning of hybridoma cultures, 11 monoclonal antibodies were further characterized. These monoclonals could be placed into five major groups based on their differential reactivities with extracts of breast tumor metastases, the surface of live mammary tumor cells in culture, and immunoper-oxidase staining of tissue sections of primary and metastatic mammary tumors. None of the antibodies bound to the surface of 14 human cell lines were derived from a variety of normal tissues, including normal mammary cells. Surface binding to mammary tumor cells by two of the monoclonal antibodies was shown to decrease during density dependent arrest, and further cell-cycle analysis demonstrated differential antibody surface binding at S phase. Prolonged exposure of mammary tumor cells to antibody showed no evidence of antigen capping or internalization.