Downstream Workflows for Intermediate Purification of Lentiviral Vectors Using Tangential Flow Filtration and AEX...

Area of Science:

• Biotechnology
• Downstream Processing
• Viral Vector Manufacturing

Background:

• The increasing demand for high-purity lentiviral vectors (LVs) necessitates scalable and cost-effective purification methods.
• Membrane chromatography, particularly anion exchange (AEX), is crucial for intermediate purification of LVs in industrial bioprocesses.
• Tangential flow filtration (TFF) can pre-treat LV harvests for concentration, buffer exchange, and impurity removal.

Purpose of the Study:

• To evaluate the integration of TFF with AEX chromatography for lentiviral vector purification.
• To compare a TFF-AEX workflow against an AEX-only workflow for LV intermediate purification.
• To assess the impact of TFF pre-treatment on AEX performance and overall impurity removal.

Main Methods:

• Investigated two workflows: AEX-only using Sartobind Convec D membranes and TFF-AEX using Hydrosart High-Performance TFF membranes followed by AEX.
• Harvested and clarified LV harvests were processed through either AEX directly or concentrated/diafiltrated via TFF before AEX loading.
• Compared dynamic binding capacity and impurity removal efficiency between the two workflows.

Main Results:

• The TFF-AEX workflow demonstrated a higher dynamic binding capacity compared to the AEX-only workflow.
• The TFF-AEX approach resulted in superior removal of impurities from the lentiviral vector harvests.
• Pre-concentration and buffer exchange using TFF optimized conditions for subsequent AEX capture.

Conclusions:

• Integrating TFF prior to AEX chromatography is an effective strategy for enhancing lentiviral vector purification.
• The TFF-AEX workflow offers improved performance metrics, including binding capacity and impurity clearance, for industrial-scale LV production.
• This combined approach provides a more robust and efficient downstream processing solution for lentiviral vectors.