Human Anti-Tetanus Toxoid Monoclonal Antibody Secreted by EBV-Transformed Human B Cells Fused with Murine Myeloma

Abstract

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.