Human anti-tetanus toxoid monoclonal antibody secreted by EBV-transformed human B cells fused with murine myeloma.
An Epstein-Barr virus (EBV)-transformed human B cell line (B6) producing anti-tetanus toxoid (TT) antibody was fused with a nonimmunoglobulin (Ig)-producing murine myeloma and selected in hypoxanthine-aminopterin-thymidine (HAT) medium containing 10(-5) M ouabain. Surviving cells were cloned by limiting dilution and confirmed as hybrids by karyotype analysis and G-11 staining. Hybridomas were stable and secreted 10-fold more anti-TT antibody (IgM kappa) than the human parental cell line. In addition, the hybridomas exhibited a markedly reduced growth requirement for serum (1% fetal calf serum). Since EBV can be used to expand rare antigen-specific B cells in the human, the technique described here may be at present the method of choice for producing human monoclonal antibodies.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000382:microbiology; D001769:Blood; D002455:Cell Division; D002459:Cell Fusion; D002460:Cell Line; D002472:Cell Transformation, Viral; D002999:Clone Cells; D003470:Culture Media; D004854:Herpesvirus 4, Human / Q000502:physiology; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007075:Immunoglobulin M / Q000096:biosynthesis; D007700:Kinetics; D051379:Mice; D009101:Multiple Myeloma; D013745:Tetanus Toxoid / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007075:Immunoglobulin M; D013745:Tetanus Toxoid