Figure 6. In vitro transduction experiments using rAAV-FLuc vectors.

(A) Transduction efficiency of the capsids, as well as AAV-DJ and AAV-LK03 capsids, on a variety of human- and nonhuman-derived cell types. Cells were transduced with the different capsid containing rAAVs that were packaging a FLuc expression cassette at a MOI of 1,000 in triplicate (with the exception of islet cells), and cell lysates were analyzed 48 hours after transduction in a luciferase activity assay. For primary human islet cells, results of 1 experiment that had been performed twice are shown. Two-fold dilutions of recombinant FLuc enzyme were used to prepare a standard curve, and raw luminescence units were calculated into luciferase molecules based on the standard curve. (B) Neutralization assay of rAAVs packaged with different capsids using dilutions of 2 different batches of pooled human immunoglobulin (IVIG). Huh-7 cells were transduced at a MOI of 100 with FLuc-expressing rAAVs that had been preincubated with different concentrations of IVIG for 1 hour at 37°C. Luciferase activity in cell lysates was measured 24 hours after transduction. Mean values of 5 replicates (obtained in 2 independent experiments) with SDs are shown for each sample. Experimental values were assessed via 2-way ANOVA using Tukey’s multiple comparisons test. Only statistically significant differences are indicated in the legend below the graph. ***P < 0.001, ****P < 0.0001.