Brazilian Archives of Biology and Technology (Jun 2014)

Production of leptospiral LipL32 antigen in Pichia pastoris and its use in an enzyme-linked immunosorbent assay

  • Leonardo Garcia Monte,
  • Fernanda Munhoz Dos Anjos Leal,
  • Daiane Drawanz Hartwig,
  • Sílvio Arruda Vasconcellos,
  • Bibiana Brihuega,
  • Odir Antonio Dellagostin,
  • Cláudia Pinho Hartleben

DOI
https://doi.org/10.1590/S1516-89132014000300007
Journal volume & issue
Vol. 57, no. 3
pp. 357 – 360

Abstract

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The production of recombinant LipL32 protein using Escherichia coli has been used extensively for the development of vaccines and diagnostic tests for leptospirosis. However, E. coli has demonstrated limitations, including low yield and lack of post-translational modifications. In this study, rLipL32 was produced in eukaryotic expression system (Pichia pastoris) and evaluated the antigen by enzyme-linked immunosorbent assay (ELISA). The yield obtained from the culture supernatant reached 270 mg/L and ELISA showed an accuracy of 95.34%. In summary, the production of rLipL32 using P. pastoris did not impair the antigenic characteristics of this antigen and ensured its use for detecting the leptospiral antibodies in swine sera.

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