Retargeting of Ad5-HVR5*7*E451Q/F35⁺⁺ to CD46 in vitro and in vivo. (A) Binding analysis was performed by surface plasmon resonance. Flow cell (Fc)1: blank immobilization; Fc2: 542RU CD46; and Fc4 455RU FX were prepared by amine coupling. Viruses at a concentration of 10¹¹ VP/mL were injected across all 4 flow cells at a flow rate of 30 μL/minute in 10 mM HEPES pH 7.4; 150 mM NaCl; 5 mM CaCl₂; 0.005% Tween 20. Representative subtracted sensorgrams (Fc2-Fc1; Fc3-Fc1; Fc4-Fc1) for each virus show the relative change in response (RU). The sensorgrams are off set for easier visualization. (B) Xgal staining of CHO-WTR and CHO-BC1 cells using 1000 viral particles of Ad5, Ad5-HVR5*7*E451Q, and Ad5-HVR5*7*E451Q/F35⁺⁺ vectors in the presence or absence of FX (*P < .05 vs PBS control). (C) Analysis of β-gal transgene expression in the liver, spleen and lung tissue after the intravascular administration of 1 × 10¹¹ vp into CD46 transgenic mice in the presence or absence of macrophage depletion. (*P < .05 vs Ad5-HVR5*7*E451Q, n = 5).