The prospect of administering genetic material using viral vectors has revolutionized the field of gene therapy. Adeno-associated virus (AAV) is among the leading and most intensively studied drug-carrying vehicles in recent advances. However, manufacturing these complex viruses while pushing efficiency and reducing costs simultaneously poses a challenging task for downstream processing. In response to the call for new solutions to replace expensive and laborious methods such as affinity chromatography, the focus is directed towards membrane chromatography. This relatively new technique aims to inherit bead-like binding capacities while eliminating diffusive mass transfer behavior restrictions given with column chromatography. This study aims to capture AAV2 from crude lysate in a single step using a mixed-mode membrane adsorber containing strong cation exchange and hydrophobic modalities. The applicability of the membrane device is examined by altering the mobile phase composition (pH, salt concentration) and elution modes (step, gradient), with an emphasis on the removal of contaminants such as host cell proteins and DNA. Additionally, a high-resolution separation was achieved at high pH, and different AAV2 species were collected. This demonstrates the ability of the mixed-mode material to distinguish between full and empty capsids. The successful isolation was evaluated using an analytical anion exchange chromatography.

Mixed-mode membrane chromatography: Primary purification of AAV2 from crude lysate reveals distinctive species behavior

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.