Characterization of secreted recombinant Toxoplasma gondii surface antigen 2 (SAG2) heterologously expressed by the yeast Pichia pastoris

Biotechnol Lett. 2008 Apr;30(4):611-8. doi: 10.1007/s10529-007-9609-x. Epub 2007 Nov 28.

Abstract

The surface antigen 2 (SAG2) gene of the protozoan parasite, Toxoplasma gondii, was cloned and extracellularly expressed in the yeast Pichia pastoris. The effectiveness of the secreted recombinant SAG2 (rSAG2-S) as a serodiagnosis reagent was assessed by western blots and ELISA. In the western blot assay, rSAG2-S reacted with all Toxoplasma-antibody positive human serum samples but not with Toxoplasma-negative samples. In the ELISA, rSAG2-S yielded sensitivity rates ranging from 80% (IgG negative, IgM positive) to 100% (IgG positive, IgM negative). In vivo experiments showed that serum from mice immunized with rSAG2-S reacted specifically with the native SAG2 of T. gondii. These mice were protected when challenged with live cells of T. gondii.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Protozoan / genetics
  • Antigens, Protozoan / immunology
  • Antigens, Protozoan / metabolism*
  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Mice
  • Mice, Inbred ICR
  • Pichia / genetics*
  • Polymerase Chain Reaction
  • Protozoan Proteins / genetics
  • Protozoan Proteins / immunology
  • Protozoan Proteins / metabolism*
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism*
  • Toxoplasma / genetics
  • Toxoplasma / immunology
  • Toxoplasma / metabolism*

Substances

  • Antigens, Protozoan
  • Protozoan Proteins
  • Recombinant Proteins
  • surface antigen P22, Toxoplasma