Production and purification of ovine anti-tetanus antibody

Comp Immunol Microbiol Infect Dis. 2005 May;28(3):167-76. doi: 10.1016/j.cimid.2005.01.001. Epub 2005 Feb 16.

Abstract

We used the ovine as bioreactor for the production and optimization of anti-tetanus toxin antibody. Four female sheep were immunized with human tetanus vaccine (TT-alum) every two weeks for 16 weeks, after which serum was collected and its titer was estimated by ELISA. The highest titer obtained was 39,000 IU ml-1. To optimize a purification protocol for ovine anti-tetanus toxin, we used four procedures; weak anion (DEAE-Sephadex), weak cation (CM-Sephadex), ammonium sulfate precipitation alone or in combination with caprylic acid. Fifty percent saturation with ammonium sulfate combined with caprylic acid gave us the highest yield of protein with specific activity and the purest Fab product.

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Antibody Specificity
  • Bioreactors
  • Chemical Precipitation
  • Chromatography, Ion Exchange / veterinary
  • Female
  • Humans
  • Immunization / veterinary*
  • Immunoglobulin Fab Fragments / biosynthesis
  • Immunoglobulin Fab Fragments / immunology
  • Sheep / immunology*
  • Tetanus / prevention & control*
  • Tetanus Antitoxin / biosynthesis*
  • Tetanus Antitoxin / blood
  • Tetanus Antitoxin / chemistry
  • Tetanus Antitoxin / pharmacology
  • Tetanus Toxoid / immunology
  • Tetanus Toxoid / isolation & purification*

Substances

  • Antibodies, Bacterial
  • Immunoglobulin Fab Fragments
  • Tetanus Antitoxin
  • Tetanus Toxoid