Anti-PorA antibodies elicited by immunization with peptides conjugated to P64k

Biochem Biophys Res Commun. 2000 Oct 14;277(1):51-4. doi: 10.1006/bbrc.2000.3623.

Abstract

To increase the humoral immune response against two cyclic synthetic peptides, derived from variable regions within the outer membrane meningococcal protein PorA (subtypes 19 and 15), we conjugated the peptides to P64k, a novel carrier protein from the same bacterium expressed in Escherichia coli. In addition, one of these peptides was restricted to a linear conformation before it was chemically coupled to the carrier. The conjugates were administered to mice in a three-dose immunization schedule, resulting in a potent anti-peptide immune response, which suggested that chemical conjugation to this carrier provided T-cell help. Antisera directed to the three conjugates reacted with Neisseria meningitidis outer membrane PorA upon immunoblot analysis. Moreover, in two out of three conjugates, the anti-peptide sera reacted with native meningococcal outer membrane vesicles in ELISA.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology*
  • Bacterial Outer Membrane Proteins / immunology
  • Bacterial Outer Membrane Proteins / metabolism*
  • Blotting, Western
  • Carrier Proteins / metabolism
  • Cross-Linking Reagents / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli
  • Immunization Schedule
  • Mice
  • Molecular Sequence Data
  • Neisseria meningitidis / immunology*
  • Peptide Fragments / chemical synthesis
  • Peptide Fragments / chemistry
  • Peptide Fragments / immunology*
  • Peptide Fragments / metabolism
  • Peptides, Cyclic / chemical synthesis
  • Peptides, Cyclic / chemistry
  • Peptides, Cyclic / immunology
  • Peptides, Cyclic / metabolism
  • Porins / chemical synthesis
  • Porins / chemistry
  • Porins / immunology*
  • Porins / metabolism*
  • Recombinant Proteins
  • T-Lymphocytes / immunology

Substances

  • Antibodies
  • Bacterial Outer Membrane Proteins
  • Carrier Proteins
  • Cross-Linking Reagents
  • Peptide Fragments
  • Peptides, Cyclic
  • Porins
  • Recombinant Proteins
  • lpdA protein, Neisseria meningitidis
  • porin protein, Neisseria