Figure 3. Injection of Adv-IK17-scFv delivers functional IK17-scFv into mouse plasma.

(A) ELISA of plasma from wild type C57BL/6 and the LDLR^−/−/Rag1^−/− mice at indicated time post-injection with 2.5 × 10¹¹ virus particles of Adv-IK17-scFv. The binding to each antigen is the average of results from two mice from each group assayed, with all determinations in the same assay. Repeat injections of Adv-IK17-scFv restored the expression of transgene in LDLR^−/−/Rag1^−/− mice, but not in C57BL/6 wt mice. (B and C) IK17-scFv plasma expression in LDLR^−/−/Rag1^−/− mice throughout 16 weeks of gene transfer in Study 2. Shown are binding to indicated antigens (in RLU/100 msec) using plasma (1:50 dilution) from Adv-IK17-scFv or Adv-EGFP injected mice one week post-injection with 10¹¹ of indicated viral particles (B), as well as the terminal blood after 16 weeks of HC-diet and 9 biweekly injections (C). (D and E) Adenovirus expressed IK17-scFv inhibits the binding of OxLDL to J774 macrophages. Fixed concentration of biotinylated MDA-LDL (1.5 µg/ml; D) or Cu-OxLDL (0.8 µg/ml; E) were incubated with indicated dilutions of pooled plasma from Adv injected animals (graphed as reciprocals, e.g. 1/100 = 0.01). Y axis indicates the biotinylated OxLDL bound to macrophages expressed as a ratio of binding in the presence of competitor (B) divided by binding in the absence of competitor (B₀). In D and E, BSA alone is used as a comparator.