Experimental & Applied Acarology

, Volume 32, Issue 1, pp 119–128

Production and biochemical characterization of the recombinant Boophilus microplus Bm95 antigen from Pichia pastoris

  • O. Boué
  • O. Farnós
  • A. González
  • R. Fernández
  • J.A. Acosta
  • R. Valdés
  • L.J. González
  • Y. Guanche
  • G. Izquierdo
  • M. Suárez
  • I. Domínguez
  • H. Machado
  • M. Rodríguez
  • R. Lleonart
Article

DOI: 10.1023/B:APPA.0000018199.87122.e4

Cite this article as:
Boué, O., Farnós, O., González, A. et al. Exp Appl Acarol (2004) 32: 119. doi:10.1023/B:APPA.0000018199.87122.e4

Abstract

The new antigen Bm95 from the cattle tick Boophilus microplus was recently isolated, cloned and expressed in the methylotrophic yeast Pichia pastoris. The recombinant protein has shown to induce protection in cattle against infestations of B. microplus under controlled and production conditions. In this paper we report the production and large-scale purification of the Bm95 protein, following a simple and cost-effective process. The antigen was obtained highly aggregated, forming particles ranging from 26 to 30 nm and with purity higher than 80%. The process yield was 0.55 g of pure Bm95 protein per liter of culture. The 98% of the primary structure of the recombinant protein was verified by mass spectrometry. Three amino acid changes in comparison with the sequence deduced from cDNA were detected by LC-MS/MS. The antigen was also obtained N-glycosylated, as previously reported for heterologous protein expression in P. pastoris.

Bm86Bm95Boophilus microplusCattle tickPichia pastoris

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • O. Boué
    • 1
  • O. Farnós
    • 1
  • A. González
    • 2
  • R. Fernández
    • 2
  • J.A. Acosta
    • 2
  • R. Valdés
    • 2
  • L.J. González
    • 3
  • Y. Guanche
    • 3
  • G. Izquierdo
    • 1
  • M. Suárez
    • 1
  • I. Domínguez
    • 4
  • H. Machado
    • 1
  • M. Rodríguez
    • 1
  • R. Lleonart
    • 1
  1. 1.Division of Mammalian Cell GeneticsCenter for Genetic Engineering and BiotechnologyHavanaCuba
  2. 2.Division of Gavac™ ProductionCenter for Genetic Engineering and BiotechnologyHavanaCuba
  3. 3.Division of Physical-ChemistryCenter for Genetic Engineering and BiotechnologyHavanaCuba
  4. 4.Division of Culture CollectionCenter for Genetic Engineering and BiotechnologyHavanaCuba