Fig. 6: Polish of full-genome AAV vectors with a hydroxyapatite column. | Gene Therapy

Fig. 6: Polish of full-genome AAV vectors with a hydroxyapatite column.

From: Large-scale purification of functional AAV particles packaging the full genome using short-term ultracentrifugation with a zonal rotor

Fig. 6: Polish of full-genome AAV vectors with a hydroxyapatite column.

Full-genome AAV vectors (#Z7) were attached to a hydroxyapatite column and eluted by sodium phosphate. UV absorbance at 260 nm and 280 nm, and viral genome titer of each fraction of chromatography without (A) or with CaCl2 (B) were evaluated. AAV genome (vg/ml), UV absorbance at 260 nm, UV absorbance at 280 nm and the conductivity of elution buffer were measured. Capsids were detected by western blotting with anti-VP1, VP2, and VP3 antibodies (C). D Recovery of rAAVs (Fr.E) polished by CHT in all fractions was determined by viral genome titer (n = 3, means ± SEM).

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