Functional and proteomic comparison of different techniques to produce equine anti-tetanus immunoglobulin F(ab')2 fragments

J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Aug 15:1092:29-39. doi: 10.1016/j.jchromb.2018.05.041. Epub 2018 May 29.

Abstract

Tetanus is still a major cause of human deaths in several developing countries. In particular, the neonatal form remains a significant public health problem. According to the World Health Organization, administration of tetanus toxoid is recommended for neonatal tetanus patients. Furthermore, tetanus antitoxin or anti-tetanus immunoglobulin (Ig) are used for mild case or intensive care. This paper discusses a novel purification technique for improving equine anti-tetanus Ig production. First, equine plasma dealt with two steps salting out with ammonium sulfate; second, ultrafiltration concentration liquid purified by one successive protein G based affinity chromatography steps; finally, the purified F(ab')2 fragments was characterized using biochemical and proteomic methods and shown to be pure and homogeneous. Compared with the original technique product, specific activity increased by 80% (about 90,000 IU/g) and recovery of F(ab')2 is approximately equal 75%. Furthermore, Proteomic profiling of total technique process is demonstrated by nano-HPLC-MS and bioinformatics analysis. New technique to produce equine anti-tetanus immunoglobulin F(ab')2 fragments from crude plasma in high quality and yield. And it also could be used for industrial amplification.

Keywords: Affinity chromatography; Anti-tetanus immunoglobulin; Equine; F(ab′)2 fragments; Purification; Tetanus antitoxin.

MeSH terms

  • Animals
  • Chromatography, Affinity
  • Horses
  • Immunoglobulin Fab Fragments / blood
  • Immunoglobulin Fab Fragments / chemistry
  • Immunoglobulin Fab Fragments / isolation & purification*
  • Immunoglobulin G / blood
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / isolation & purification*
  • Mass Spectrometry
  • Proteomics
  • Tetanus / immunology*
  • Tetanus Antitoxin / isolation & purification*

Substances

  • Immunoglobulin Fab Fragments
  • Immunoglobulin G
  • Tetanus Antitoxin