Isolation of human monoclonal antibodies binding to B fragment of diphtheria toxin.
Human tonsillar lymphocytes immunized in vitro with diphtheria toxoid were fused with the heteromyeloma cell line SP2/HPT. Hybrids were selected on hypoxanthine-thymidine aminopterin (HAT) medium and assayed for antibody production both by ELISA and Western blotting. Four stable hybridoma cell lines producing 3-7 micrograms/mL of human IgG1 antibodies were isolated. In Western blotting studies these antibodies show binding to the B fragment of diphtheria toxin. Cells from the subclone P2-13/G35 were injected i.p. in pristane-primed nude mice, and ascites fluid was collected after 4-6 weeks. The monoclonal antibody neutralized the cytotoxicity of diphtheria toxin in the Vero cell assay system. This human monoclonal antibody may have a potential in the immunotherapy of diphtheric disease.
Gupta
K C
KC
Ryerson Polytechnical Institute, Toronto, Canada.
Agha
R
R
Santos
E
E
Brodeur
B R
BR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D015153:Blotting, Western; D002459:Cell Fusion; D003601:Cytotoxicity Tests, Immunologic; D004167:Diphtheria Toxin / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D066298:In Vitro Techniques; D008214:Lymphocytes; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009101:Multiple Myeloma
D000911:Antibodies, Monoclonal; D004167:Diphtheria Toxin; D007074:Immunoglobulin G