The serum albumin-binding region of streptococcal protein G: a bacterial fusion partner with carrier-related properties

J Immunol Methods. 1997 Feb 14;201(1):115-23. doi: 10.1016/s0022-1759(96)00217-7.

Abstract

In this study, we have explored the use of the serum albumin-binding region (BB) from streptococcal protein G (SpG) as a bacterial fusion partner for production of peptide immunogens. The fusion protein BB-M3, containing BB and repeated structures from the Plasmodium falciparum malaria antigen Pf155/RESA, was efficiently purified from Escherichia coli culture supernatants by affinity chromatography using BB as an affinity tag. Rabbits immunized with BB-M3 in Freund's adjuvant produced high levels of antibodies which reacted with both M3 and BB in ELISA and stained intact Pf155/RESA in the membrane of infected erythrocytes. These antibody levels were sustained for more than 30 weeks. BB-M3 also induced antibody responses to M3, BB and intact Pf155/RESA in a number of mouse strains, including several strains which are non-responders to the malaria sequences. In the latter mice, however, BB-M3 only activated BB-specific T cells, suggesting that BB has ability to provide carrier-related T cell help for antibody production. Moreover, the minimal albumin-binding motif of SpG, containing only 46 amino acids, was immunogenic in both B10.BR, B10.D2 and C57BL/6 mice (H-2k, H-2d and H-2b, respectively). These results indicate that BB has both affinity tag and carrier-related properties and suggest that fusion proteins containing BB can be efficient tools for the generation of antibody responses to peptides which are weak immunogens.

MeSH terms

  • Animals
  • Antibodies, Protozoan / biosynthesis*
  • Bacterial Proteins / chemistry*
  • Haplotypes
  • Lymphocyte Activation
  • Malaria Vaccines / chemistry*
  • Mice
  • Mice, Inbred Strains / immunology
  • Peptides / immunology
  • Protein Binding
  • Rabbits
  • Recombinant Fusion Proteins / immunology*
  • Serum Albumin / chemistry*
  • T-Lymphocytes / immunology*
  • Vaccines, Synthetic / immunology*

Substances

  • Antibodies, Protozoan
  • Bacterial Proteins
  • IgG Fc-binding protein, Streptococcus
  • Malaria Vaccines
  • Peptides
  • Recombinant Fusion Proteins
  • Serum Albumin
  • Vaccines, Synthetic