Monday, July 25, 2011
Grand Ballroom, 5th fl (Sheraton New Orleans)
The rapid production of vaccines at high titer and potency would greatly facilitate addressing national vaccine needs. The rapid fermentation of E. coli for the over expression of recombinant protein vaccine products provides the flexibility and capacity to meet this ongoing need. Recombinant fusion-protein vaccine products that include the ability to stimulate Toll-Like Receptor 5 (TLR5) have been demonstrated to show significantly higher potency than simple recombinant protein. In this project Salmonella typhimurium FliC (flagellin monomer) capable of stimulating TLR5 was used as the platform for fusion with the target antigen. Marburg Virus (MARV) is a member of the Filovirus family; it is recognized as a Bioterrorism agent which currently does not have a licensed vaccine. GP132 is a 15mer oligopeptide from the surface glycoprotein of MARV, and previously shown to provide protection from MARV challenge in animal studies. A recombinant full-length FliC producing E. coli clone was developed as an initial platform for the production of purified FliC and the FliC MARV linked vaccine product. A full-length FliC producing clone was grown using a Biostat C 20L vessel and the protein product purified. Results for the generation of the full-length FliC producing clone, fermentation, and purification will be presented. The FliC containing plasmid was also used as the foundation for the generation of the phase II product where GP132 was fused to the c-terminal region of full-length FliC. The preliminary results of the generation of this clone will also be presented.