Fouling-Free Inertial Microfluidic Cell Clarification for Lentivirus Manufacturing

Area of Science:

• Biotechnology
• Gene Therapy Manufacturing
• Process Engineering

Background:

• Lentiviral vectors (LVVs) are crucial for gene therapy, but their large-scale production is hindered by inefficient harvest clarification.
• Conventional depth filtration methods often lead to significant product loss, impacting manufacturing yields.

Purpose of the Study:

• To develop and evaluate a fouling-free microfluidic cell clarification (MCC) system for high-efficiency lentiviral vector (LVV) recovery.
• To establish a quantitative relationship between clarification efficiency, throughput, and operational parameters for LVV manufacturing.

Main Methods:

• A microfluidic cell clarification (MCC) system was designed and optimized by controlling harvest flow rate and cell density.
• Clarification efficiency and throughput were quantitatively assessed under various operating conditions.
• Performance was benchmarked against conventional cellulose depth filtration processes.

Main Results:

• The MCC system achieved over 95% LVV product recovery and over 95% cell removal efficiency under optimized conditions.
• LVV recovery closely matched theoretical models, indicating minimal product loss within the device.
• The MCC demonstrated superior recovery compared to a benchmark depth filtration process across multiple batches.
• Scalability was confirmed up to 1.5 L/h flow rate without compromising clarification efficiency.

Conclusions:

• The fouling-free MCC system offers a highly efficient and scalable solution for lentiviral vector clarification in biomanufacturing.
• This technology presents a significant advancement over conventional filtration, minimizing product loss and improving yields.
• The MCC platform shows broad applicability for clarifying cell-based therapeutic products, providing a continuous, low-shear alternative.