Figure 1.
Validation of incorporation of sdAb-targeted chimeric fiber protein in CRAds. (a) Simplified schematic overview of adenovirus vectors used in this study. Only relevant genomic regions are shown. (b) Validation of incorporation of fiber-fibritin-sdAb into the viral capsid using western blot analysis. Equal amounts (5 × 109 vp) of purified viral particles from Ad5, Ad.CXCR4E1 and Ad.CXCR4E1.B2 were loaded in sample buffer in each lane without (lane 1, 3, and 6) or with boiling (lane 2, 4, and 7). Proteins were separated on a SDS-PAGE gel followed by western blot transfer to a PVDF membrane. Fiber protein expression was detected using antifiber mAb. Predicted molecular weight (MW) of wild-type Ad5 fiber monomers is 61.6 kDa and MW 67.7 kDa for fiber-fibritin-sdAb. One representative of three different experiments is shown. B, boiled; LITR, left inverted terminal repeat; M, marker; PVDF, polyvinylidene difluoride; RITR, right inverted terminal repeat; U, unboiled; ΔE1, E1 deleted.