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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 96, No. s1, 2001, pp. 131-135
Bioline Code: oc01163
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 96, No. s1, 2001, pp. 131-135

 en r-Sm14 - pRSETA Efficacy in Experimental Animals
Celso Raul Romero Ramos; Mônica Magno Vilar; Ana Lúcia Tabet Oller Nascimento; Paulo Lee Ho; Nilton Thaumaturgo; Ricardo Edelenyi; Marília Almeida; Waldely de Oliveira Dias; Catia Maria Diogo & Miriam Tendler

Abstract

Previous studies carried out with Sm14 in experimental vaccination against Schistosoma mansoni check for this species in other resources or Fasciola hepatica check for this species in other resources infections were performed with recombinant Sm14 (rSm14) produced in Escherichia coli check for this species in other resources by the pGEMEX system (Promega). The rSm14 was expressed as a 40 kDa fusion protein with the major bacteriophage T7 capsid protein. Vaccination experiments with this rSm14 in animal models resulted in consistent high protective activity against S. mansoni cercariae challenge and enabled rSm14 to be included among the vaccine antigens endorsed by the World Health Organization for phase I/II clinical trials. Since the preparation of pGEMEX based rSm14 is time consuming and results in low yield for large scale production, we have tested other E. coli expression systems which would be more suitable for scale up and downstream processing. We expressed two different 6XHis-tagged Sm14 fusion proteins in a T7 promoter based plasmids. The 6XHis-tag fusions allowed rapid purification of the recombinant proteins through a Ni+2-charged resin. The resulted recombinant 18 and 16 kDa proteins were recognized by anti-Sm14 antibodies and also by antiserum against adult S. mansoni soluble secreted/excreted proteins in Western-Blot. Both proteins were also protective against S. mansoni cercariae infection to the same extent as the rSm14 expressed by the pGEMEX system.

Keywords
Schistosoma mansoni, fatty acid binding protein, anti-helminth vaccine

 
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