Lentiviral Vector Purification Using Nanofiber Ion-Exchange Chromatography.

Area of Science:

• Biotechnology
• Gene Therapy
• Virology

Background:

• Lentiviral vectors (LVs) are crucial for cell and gene therapies, enabling long-term gene expression in various cell types.
• Current production methods yield low titers (10^5-10^7 TU/mL), necessitating efficient concentration and purification strategies.
• Existing purification methods often struggle with scalability and impurity removal.

Purpose of the Study:

• To develop a scalable and efficient method for concentrating and purifying lentiviral vectors (LVs).
• To simplify upstream processing for LV production using a specialized packaging cell line.
• To evaluate a novel stationary phase for high-capacity LV capture and impurity removal.

Main Methods:

• Utilized the WinPac-RD-HV packaging cell line for LV production.
• Developed a direct capture method employing ion-exchange chromatography with cellulose nanofibers.
• Assessed the concentration factor, impurity removal (host cell proteins), and functional vector yield.

Main Results:

• Achieved a 100-fold concentration of lentiviral vectors.
• Demonstrated a two-log reduction in host cell protein impurities.
• Maintained a high yield of functional vector, up to 90%.

Conclusions:

• The developed ion-exchange chromatography method using cellulose nanofibers is a scalable and effective approach for LV concentration and purification.
• This novel method significantly improves LV production efficiency and purity, supporting advancements in cell and gene therapy.
• The high surface area and accessibility of the cellulose nanofiber stationary phase offer superior performance compared to traditional methods.