ion exchange column  (Bio-Rad)

Journal: Journal of Biological Engineering

Article Title: Advanced biomanufacturing and evaluation of adeno-associated virus

doi: 10.1186/s13036-024-00409-4

Figure Lengend Snippet: 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

Article Snippet: To develop a universal primary purification method for multiple AAV serotypes (AAV2, 5, DJ and DJ/8), Bio-Rad NGC system equipped with four chromatography columns, including EconoFit Nuvia aPrime 4A ion exchange (IEX, anionic) column (Bio-Rad), Foresight Nuvia HPQ anionic column (Bio-Rad), HiTrap Q Sepharose XL anionic column (Cytiva, Marlborough, MA, USA), and HiTrap AVB Sepharose affinity column (Cytiva), for AAV purification.

Techniques: Purification, Liquid Chromatography, Lysis

Journal: Journal of Biological Engineering

Article Title: Advanced biomanufacturing and evaluation of adeno-associated virus

doi: 10.1186/s13036-024-00409-4

Figure Lengend Snippet: 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

Article Snippet: Multiple commercial columns for AAV purification have been evaluated in this study (Table ), including Cytiva HiTrap Q Sepharose XL strong anion exchange column, Cytiva Sepharose Fast Flow anion exchange column, Cytiva HiTrap AVB Sepharose column, Bio-Rad Foresight Nuvia HPQ column, and Bio-Rad EconoFit Nuria aPrime 4A.

Techniques: Purification, Liquid Chromatography, Lysis