Figure 4.

Purification of the rAAV2/8 vectors using the CEX-AEX process. (A–C) Chromatograms obtained with cell pellet materials in only two steps on the CEX (A) and AEX (B) columns. (A) CEX with a linear buffer B gradient. (B) AEX with a linear buffer B gradient. Magnification view of the AEX chromatogram shown in B. Each chromatogram shows the respective UV signal in blue (scale on the left Y-axis) and the concentration of buffer B depicted as a dashed green line (scale on the right). (C) Graph showing the yield (total vector genomes, vg) for the rAAV2/8 vectors obtained in CEX-AEX with the material harvested by lysis of the cell pellet. The residual HEK cell DNA amount is shown in (D). Due to matrix effects in the qPCR, the starting amount cannot be shown here. AAV, recombinant adeno-associated virus; AC, affinity chromatography; AEX, anion exchange chromatography; CEX, cation exchange chromatography.