Fig. (2) Validation and characterization of the chimeric vector. (A) To demonstrate the hexon switch on Ad5, PCR was utilized to detect the hexon3 gene from viral genomes of vectors. Lanes 1 through 4 are ordered as follows: Ad5, Ad3, Ad5H3 and Ad5H3-HVR2-His. (B) To demonstrate the incorporation of a His₆ tag into hexon3 on chimeric Ad5H3, western-blot analysis was performed. Lanes 1 through 3 in panel B are as follows: Ad3, Ad5H3, and Ad5H3-HVR2-His. (C) In order to verify His₆ surface display on the vectors, ELISA assays were performed. (D) Coommassie blue staining analyses was carried out to demonstrate that the “Antigen Capsid-Incorporation” strategy would not affect the adenovirus structural integrity. Lanes 1 through 4 are ordered as follows: Ad5, Ad3, Ad5H3 and Ad5H3-HVR2-His.