Membrane adsorber design for lentiviral vector recovery

Area of Science:

• Biotechnology
• Bioprocessing
• Chromatography

Background:

• Lentiviral vectors (LVs) are crucial for cell and gene therapies, but large-scale production faces challenges with low product recovery.
• Anion-exchange chromatography (AIEX) is a key step for LV capture but often causes significant product loss due to irreversible binding.

Purpose of the Study:

• To develop and evaluate novel AIEX membrane structures designed to reduce LV interaction strength and minimize product loss.
• To improve functional recovery of lentiviral vectors during downstream processing.

Main Methods:

• Four AIEX membrane prototypes with varying ligand densities and pore sizes (1.05 and 1.32 μm) were fabricated using Q and D chemistries.
• Prototypes were tested for their ability to minimize irreversible LV binding and improve recovery rates.
• Performance was assessed by measuring total particle recovery, elution profiles, and functional recovery of LVs encoding GFP and CAR transgenes.

Main Results:

• The novel AIEX membranes minimized product loss from irreversible binding, achieving high total particle recoveries (>80%) even after extended adsorption times.
• Narrower elution ranges were observed, with LVs eluting at lower salt concentrations (<450 mM NaCl).
• The DR2 prototype demonstrated a ~3-fold higher functional recovery for GFP and CAR LVs compared to standard Q-membranes, and a 3.5-fold improvement at drug substance scale (43% vs. 12%).

Conclusions:

• Optimized AIEX membrane design, by tuning ligand density and pore size, significantly enhances lentiviral vector recovery in downstream processing.
• These findings offer a promising solution to improve the efficiency and yield of LV production for therapeutic applications.
• The developed adsorbents represent a significant advancement in overcoming critical bottlenecks in cell and gene therapy manufacturing.