title: Neutralization of tetanus toxin by human monoclonal antibodies directed against tetanus toxin fragment C.
abstract: Two hybridomas (designated 143 and 147) producing human monoclonal antibodies (IgG1) directed against tetanus toxin were established by fusion of Epstein-Barr virus transformed human peripheral B lymphocytes with the heteromyeloma SPAM-8. The hybridomas produced antibodies in concentrations of approx. 3.5 micrograms/ml (hybridoma 143) and 6.4 micrograms/ml (hybridoma 147) using conventional flask cultures and 33.9 micrograms/ml and 36.2 micrograms/ml, respectively, in dialysis cultures. The antibodies were shown to react with tetanus toxin, toxoid and fragment C in ELISA, and reactivity with tetanus toxin and fragment C was confirmed in SDS-polyacrylamide gel electrophoresis followed by Western blots. The antibody binding sites were located to two different epitopes of fragment C as shown in a competition assay using biotinylated antibodies. Furthermore, binding of both antibodies to fragment C was inhibited by the addition of the receptor-associated ganglioside GT1b. Neutralization of tetanus toxin in concentrations equivalent to 100-120 IU per mg of antibody was observed for both antibodies in a mouse protection assay.
authors.affiliation: Department of Bacteriology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: B
authors.initials: B
authors.lastname: Gustafsson
authors.forename: E
authors.initials: E
authors.lastname: Whitmore
authors.forename: M
authors.initials: M
authors.lastname: Tiru
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D013744:Tetanus Toxin / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D013744:Tetanus Toxin