title: Expression of tetanus toxin subfragments in vitro and characterization of epitopes.
abstract: To define epitopes of tetanus toxin, we compared four different in vitro systems in terms of their ability to produce tetanus toxin-specific subfragments from cloned DNA. A transcription-translation system developed from a nontoxigenic strain of Clostridium tetani was found to yield predominantly full-sized peptides. Such peptides were used to map six different epitopes for eight monoclonal antibodies. The toxin-neutralizing properties of the antibodies were determined in an in vitro assay, based on the toxin-mediated inhibition of norepinephrine release from rat brain particles. Two monoclonal antibodies recognizing epitopes within the regions Ser-744 to Ser-864 and Ile-1224 to Asp-1315 could neutralize the toxin. A third nonneutralizing antibody was shown to recognize the synthetic peptide Phe-947 to Glu-967 derived from the tetanus toxin sequence. This peptide contains a human T-cell epitope.
authors: Institut für Medizinische Virologie, Justus Liebig Universität, Giessen, Federal Republic of Germany.
authors: B
authors: B
authors: Andersen-Beckh
authors: T
authors: T
authors: Binz
authors: H
authors: H
authors: Kurazono
authors: T
authors: T
authors: Mayer
authors: U
authors: U
authors: Eisel
authors: H
authors: H
authors: Niemann
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002474:Cell-Free System; D003001:Cloning, Molecular; D003017:Clostridium tetani / Q000235:genetics* / Q000276:immunology; D000939:Epitopes; D066298:In Vitro Techniques; D009638:Norepinephrine / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D015183:Restriction Mapping; D013744:Tetanus Toxin / Q000235:genetics* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D013744:Tetanus Toxin; D009638:Norepinephrine