r-Sm14 - pRSETA efficacy in experimental animals.
Previous studies carried out with Sm14 in experimental vaccination against Schistosoma mansoni or Fasciola hepatica infections were performed with recombinant Sm14 (rSm14) produced in Escherichia coli 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.
Ramos
C R
CR
Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brasil.
Vilar
M M
MM
Nascimento
A L
AL
Ho
P L
PL
Thaumaturgo
N
N
Edelenyi
R
R
Almeida
M
M
Dias
W O
WO
Diogo
C M
CM
Tendler
M
M
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D015153:Blotting, Western; D002352:Carrier Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000276:immunology / Q000378:metabolism; D050612:Fatty Acid Transport Proteins; D005260:Female; D015801:Helminth Proteins / Q000276:immunology* / Q000378:metabolism; D026901:Membrane Transport Proteins; D051379:Mice; D023421:Models, Animal; D010957:Plasmids / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012550:Schistosoma mansoni / Q000276:immunology* / Q000378:metabolism; D014611:Vaccination; D014612:Vaccines / Q000276:immunology*
D000909:Antibodies, Helminth; D002352:Carrier Proteins; D050612:Fatty Acid Transport Proteins; D015801:Helminth Proteins; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; D014612:Vaccines; C068709:SM14 protein, Schistosoma mansoni