Fig. 5

Development and optimization of anion exchange purification using liquid chromatography (LC). The 80–140 mL of cell lysis from 20–100 mL of VPC pellet was loaded to the 1-mL or 5-mL columns. The representative LC profile of AAV-DJ8 was described here but four serotypes of AAV2, 5, DJ and DJ8 were tested using the developed purification strategy. Equilibration buffer: 25 mM Tris–HCl, pH 9.0. Elution Buffer A: 25 mM Tris–HCl, 20 mM NaCl, pH 9.0. Elution buffer B: 25 mM Tris–HCl, 1 M NaCl, pH 9.0. Flow rate: 1.0 mL/min. A Stepwise elution of AAV-DJ8, 80-mL AAV lysis from 20-mL VPC pellet, in 1-mL EconoFit Nuvia aPrime 4A column. B Stepwise elution of AAV-DJ8, 140-mL AAV lysis from 100-mL VPC pellet, in 1-mL EconoFit Nuvia aPrime 4A column. C Stepwise elution of AAV-DJ8, 100-mL AAV lysis from 50-mL VPC pellet, using 1-mL Foresight Nuvia HPQ column, which can be scaled up from 1-mL column to 5-mL and 25-mL columns. D Stepwise elution of AAV-DJ8, 100-mL AAV lysis from 50-mL VPC pellet, using 5-mL Foresight Nuvia HPQ column