3.1 Generation and characterization of VSV/RABV-GP
To generate VSV/RABV-GP, the GP gene of RABV from the Era strain was
cloned into the pcDNA3.1 vector and is designated as pcDNA3.1-RABV-GP.
The VSV/RABV-GP pseudo-typed virus was generated in HEK293T cells, as
described previously (Park et al., 2016) . The incorporation of
RABV-GP was verified by sandwich ELISA. As shown in Figure 1A, the
absorbance at 450nm in wells containing VSV/RABV-GP was significantly
higher than that in wells containing VSV/VSV-GP or background. To
evaluate whether the recombinant GP was antigenically similar to wild
type RABV-GP, the response to neutralizing antibodies in BHK21 cells was
evaluated. As shown in Figure 1B, VSV/RABV-GP transduction was
significantly reduced by neutralizing antibodies, but it was not
affected by the off-target antibody (anti-C9), which was used as a
control. As expected, no inhibitory activity was detected against
VSV/VSV-GP, the negative control. These results clearly confirmed that
the RABV-GP was incorporated correctly on the surface of the VSV
pseudotyped virus and displayed critical neutralizing epitopes such that
it induced potential neutralizing antibodies against RABV.