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

E A Green, C Botting, H M Webb, T R Hirst, R E Randall

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)949-58
Number of pages10
JournalVaccine
Volume14
Issue number10
Publication statusPublished - 1996

Keywords

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Antibody Formation
  • Antigen-Antibody Reactions
  • Bacterial Toxins
  • Base Sequence
  • Binding Sites, Antibody
  • Enterotoxins
  • Escherichia coli Proteins
  • G(M1) Ganglioside
  • Immunity, Cellular
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Simian immunodeficiency virus
  • Vaccines, Synthetic

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