Construction, purification and immunogenicity of antigen-antibody-LTB complexes.
An oligonucleotide, encoding a short epitope peptide tag, termed Pk, was inserted at the 3'-end of the gene coding B-subunit of Escherichia coli heat-labile enterotoxin (LTB). The presence of the Pk epitope on LTB-Pk was used to construct novel macromolecular assemblies comprising LTB-Pk, an anti-Pk mAb, (mAb SV5-P-k) and Pk-linked recombinant SIV proteins. The 1:1:1 stoichiometry of such complexes was ensured by binding LTB-Pk to one arm of mAb SV5-P-k and an SIV-Pk antigen to the other arm of the antibody. Such SIV-mAb-LTB macromolecular complexes bound to GM1-ganglioside in vitro, and when immunized systemically into mice were highly immunogenic, inducing both humoral and cell-mediated responses to the recombinant SIV antigens.
Green
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School of Biological and Medical Sciences, University of St. Andrews, Fife, UK.
Botting
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Webb
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Hirst
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Randall
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000917:Antibody Formation; D000937:Antigen-Antibody Reactions; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology*; D001483:Base Sequence; D001666:Binding Sites, Antibody; D004768:Enterotoxins / Q000235:genetics* / Q000276:immunology*; D029968:Escherichia coli Proteins; D005677:G(M1) Ganglioside / Q000276:immunology; D007111:Immunity, Cellular; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015302:Simian Immunodeficiency Virus / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D014614:Vaccines, Synthetic; D005677:G(M1) Ganglioside; C027538:heat-labile enterotoxin, E coli