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Bacterial Expression System> | … Membranes were blocked with 5% non-fat dry milk (NFDM) in Tris-buffered saline with 0.05% Tween-20 (TBST) for 1 h at room temperature, then for 1 h with anti-His mAb (GenScript, Piscataway, NJ) diluted at 1:500 in TBST … | Get A Quote |
In this study, an alphavirus vector platform was used to deliver replicon particles (RPs) expressing African swine fever virus (ASFV) antigens to swine. Alphavirus RPs expressing ASFV p30 (RP-30), p54 (RP-54) or pHA-72 (RP-sHA-p72) antigens were constructed and tested for expression in Vero cells and for immunogenicity in pigs. RP-30 showed the highest expression in Vero cells and was the most immunogenic in pigs, followed by RP-54 and RP-sHA-p72. Pigs primed with two doses of the RP-30 construct were then boosted with a naturally attenuated ASFV isolate, OURT88/3. Mapping of p30 identified an immunodominant region within the amino acid residues 111-130. However, the principal effect of the prime-boost was enha... More