Laboratory Scale Production of Recombinant Haa86 Tick Protein in Pichia pastoris and in Escherichia coli System

Methods Mol Biol. 2016:1404:459-482. doi: 10.1007/978-1-4939-3389-1_30.

Abstract

The commercial recombinant Bm86-based vaccines against Rhipicephalus (Boophilus) microplus in Australia (TickGARD™, TickGARD plus™) and in Cuba (Gavac™) provided significant impetus to researchers globally to work on anti-tick vaccines. The Bm86 homologue of Hyalomma anatolicum anatolicum Izatnagar isolate, rHaa86, is considered a potent vaccine candidate against Hyalomma tick species, transmitting animal and human diseases. The two expression systems, prokaryotic, using bacterial expression plasmid vectors and Escherichia coli, and eukaryotic, using methylotrophic yeast Pichia pastoris and yeast expression plasmid vectors, are used in the laboratory level production of rHaa86. Unlike proteins expressed in prokaryotic system, eukaryotic system-expressed proteins are glycosylated and may be a requisite for proper immunogenicity. Here, the protocol for laboratory scale expression of rHaa86 antigen in E. coli and P. pastoris for development of an anti-Hyalomma tick vaccine is described.

Keywords: Anti-Hyalomma vaccine; Eukaryotic expression; Laboratory scale production; Prokaryotic expression; rHaa86.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arthropod Proteins / biosynthesis*
  • Arthropod Proteins / genetics
  • Arthropod Proteins / isolation & purification
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Escherichia coli / genetics*
  • Genetic Engineering / methods*
  • Ixodidae*
  • Laboratories*
  • Pichia / genetics*
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Sequence Analysis, DNA

Substances

  • Arthropod Proteins
  • DNA, Complementary
  • Recombinant Proteins