A versatile system for the production of recombinant chimeric peptides

K. A.L. De Smet, H. M. Vordermeier, J. Ivanyi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Production of chimeric and multimeric peptides is of interest for the analysis of topographic relationships between T and B cell stimulatory epitopes. Recombinant DNA technology has certain advantages over conventional chemical peptide synthesis for the production of peptide constructs of large size (more than 40 amino acid residues). We describe a methodology which is versatile and independent of the expression vector used because it only relies on the incorporation of appropriate restriction enzyme sites in oligonucleotides. The method was verified using two 20mer sequences from the 38 kDa antigen of M. tuberculosis. Peptide 201-220, containing an antibody binding linear epitope, has been made immunogenic in vivo when combined with T cell stimulatory peptide 350-369 in a chimeric peptide. The results demonstrate that a distinct orientation of the constituent peptides was essential for achieving optimal immunogenicity.

Original languageEnglish
Pages (from-to)243-250
Number of pages8
JournalJournal of Immunological Methods
Volume177
Issue number1-2
DOIs
Publication statusPublished - 28 Dec 1994

Keywords

  • B-T cell cooperation
  • Chimeric peptide
  • Expression vector
  • Recombinant peptide
  • Subunit vaccine
  • Amino Acid Sequence
  • Mycobacterium tuberculosis/immunology
  • Lymphocyte Activation
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Cloning, Molecular/methods
  • Male
  • Epitopes
  • Animals
  • Antigens, Bacterial/chemistry
  • Recombinant Fusion Proteins/biosynthesis
  • Base Sequence
  • Antibody Formation
  • Female
  • Mice
  • Peptides/immunology

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