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Construction, purification and immunogenicity of antigen-antibody-LTB complexes

By E.A. Green, Catherine Botting, Helen M. Webb, Timothy R. Hirst and Richard Edward Randall

Abstract

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

Topics: R
Publisher: Butterworth-heinemann ltd
Year: 1996
DOI identifier: 10.1016/0264-410x(96)00039-4
OAI identifier: oai:kar.kent.ac.uk:19214
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