Microbial Cell Factories · 2010 · 27 citations · 24 references
Veterinary MedicineBiotechnologyInfectious DiseasesHost-pathogen InteractionsProtein EngineeringVaccinologyPathogen RecognitionRecombinant Protein VaccinesRecombinant Vaccine DevelopmentSubunit VaccinesVaccine Antigen DesignVaccine Antigen DiscoveryEscherichia ColiLeptospirosis VaccinesPurificationPurification Of Vaccine AntigensRecombinant Fusion ProteinsSubunit Vaccine ProductionVaccine Antigen Expression SystemsYeast-based Production
BACKGROUND: Leptospirosis, a zoonosis caused by Leptospira spp., is recognized as an emergent infectious disease. Due to the lack of adequate diagnostic tools, vaccines are an attractive intervention strategy. Recombinant proteins produced in Escherichia coli have demonstrated promising results, albeit with variable efficacy. Pichia pastoris is an alternative host with several advantages for the production of recombinant proteins. RESULTS: The vaccine candidates LigANI and LipL32 were cloned and expressed in P. pastoris as secreted proteins. Large-scale expression resulted in a yield of 276 mg/L for LigANI and 285 mg/L for LipL32. The recombinant proteins were glycosylated and were recognized by antibodies present in the sera of patients with severe leptospirosis. CONCLUSIONS: The expression of LigANI and LipL32 in P. pastoris resulted in a significant increase in yield compared to expression in E. coli. In addition, the proteins were secreted, allowing for easy purification, and retained the antigenic characteristics of the native proteins, demonstrating their potential application as subunit vaccine candidates.
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Urban epidemic of severe leptospirosis in Brazil
Albert I. Ko, Mitermayer Galvão dos Reis, Cibele Maria Ribeiro Dourado et al. · The Lancet · 1999 · 765 citations