Fig. 5

Analysis of serum from immunised mice. a Titration of the mouse antisera produced Zera®-VP2ep (blue line), Zera®-VP2 (pink line) and DPBS (negative control, grey line) vaccine candidates as well as titration of positive control sheep serum produced against BTV-8 VLPs [20] to validate the indirect ELISA (b). The markers indicate the mean value of triplicate samples from both animal experiments, and error bars indicate the standard deviation. (c) Western blot detection of the E. coli-expressed VP2 fusion protein with 1:100 dilution of pooled mice sera from animals vaccinated with the Zera®-VP2ep and Zera®-VP2 vaccines. Lane M represents the molecular weight marker. The negative control (−) was performed with no primary antibody and sheep serum obtained from BTV-8 VLP vaccinated sheep [20] was used as the positive control (+). Lanes PB and FB represent the pre-and final bleed sera respectively. The white arrow indicates the E. coli-expressed VP2 fusion protein at ~163 kDa. PB – prebleed; FB – final bleed