A novel adjuvant-free H fusion system for the production of recombinant immunogens in Escherichia coli: Its application to a 12 kDa antigen from Cryptosporidium parvum

Bioengineered. 2013 Nov-Dec;4(6):413-9. doi: 10.4161/bioe.26003. Epub 2013 Aug 9.

Abstract

The production of recombinant antigens in Escherichia coli and specific polyclonal antibodies for diagnosis and therapy is still a challenge for world-wide researchers. Several different strategies have been explored to improve both antigen and antibody production, all of them depending on a successful expression and immunogenicity of the antigen. Gene fusion technology attempted to address these challenges: fusion partners have been applied to optimize recombinant antigen production in E. coli, and to increase protein immunogenicity. Taking a 12-kDa surface adhesion antigen from Cryptosporidium parvum (CP12) by example, the novel H fusion partner was presented in this work as an attractive option for the development of recombinant immunogens and its adjuvant-free immunization. The H tag (of only 1 kDa) efficiently triggered a CP12-specific immune response, and it also improved the immunization procedure without requiring co-administration of adjuvants. Moreover, polyclonal antibodies raised against the HCP12 fusion antigen detected native antigen structures displayed on the surface of C. parvum oocysts. The H tag proved to be an advanced strategy and promising technology for the diagnosis and therapy of C. parvum infections in animals and humans, allowing a rapid and simple recombinant production of the CP12 antigen.

Keywords: CP12; Cryptosporidium; antibody production; free-adjuvant immunization; immunogens; novel fusion partner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Protozoan / blood
  • Antigens, Protozoan / biosynthesis*
  • Base Sequence
  • Cryptosporidium parvum / genetics
  • DNA, Protozoan / genetics
  • DNA, Protozoan / isolation & purification
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Fusion*
  • Mice
  • Molecular Sequence Data
  • Oocysts / immunology
  • Recombinant Proteins / biosynthesis*

Substances

  • Antibodies, Protozoan
  • Antigens, Protozoan
  • DNA, Protozoan
  • Recombinant Proteins