Expression of truncated CTB::VP60 in tobacco exhibited no immunogenicity in rabbits.

Despite several optimizations, the production of CTB::VP60 antigen fusion proteins in tobacco is still very low. This might be due to the size of the fusion partner VP60 (579 aa). Hence, two different N-terminal truncations of VP60 were fused to CTB, either with or without an ER retention signal. CTB::VP60 expression levels, in vitro and in vivo antigenicity and immunogenicity were analyzed in plants carrying one of four different transgenes. Only one of the truncated CTB::VP60 fusions (365 aa) directed to the endoplasmic reticulum led to similar but not enhanced expression levels as compared to the complete protein in tobacco and possessed similar in vitro antigenicity. In contrast to the complete protein, no anti-VP60-specific antibodies were induced in rabbits after the intramuscular application of plant extracts containing the truncated protein.