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

Brazilian Archives of Biology and Technology

Leonardo Garcia Monte²  Fernanda Munhoz Dos Anjos Leal²  Daiane Drawanz Hartwig¹  Sílvio Arruda Vasconcellos¹  Bibiana Brihuega¹  Odir Antonio Dellagostin¹  Cláudia Pinho Hartleben² 

[1] ,Universidade Federal de Pelotas Centro de Desenvolvimento Tecnológico Laboratório de ImunodiagnósticoPelotas RS ,Brasil

关键词: Pichia pastoris;    LipL32;    ELISA;    Leptospirosis;   

DOI  :  10.1590/S1516-89132014005000001

来源: SciELO

【 摘 要 】

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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