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. 2021 Aug 3;22(15):8355. doi: 10.3390/ijms22158355

Figure 3.

Figure 3

Infectivity and heparin binding differed amongst rAAV-2E3 variants. (A) Screen of infectivity of rAAV-2E3 capsid variants in comparison to AAV2 was performed on selected human and murine cell lines. Viral particles were incubated with cells (VG/cell 150,000) for three days followed by flow cytometry measurement of GFP expression. (B) HEK 293 cells infected with decreasing VG/cell (150,000, 50,000, 15,000, and 5000) of AAV2 in comparison to rAAV-2E3 capsid variants. Images of GFP expression were taken after 3 days of infection (scalebar 100 µm). (C) Analysis of rAAV-2E3 serotype interactions with heparin-agarose in comparison to AAV2. Equal amounts of rAAV-2E3 particles were loaded to heparin columns, fractions of flow through, wash, and elution were collected, and viral genomes were measured by ddPCR. The data were normalized versus the total amount of loaded viral genomes. Data represent the mean + SEM of two independent experiments. One-way ANOVA compared to AAV2 flow through, uncorrected Fisher’s LSD, *** p < 0.001, ns = non-significant.