Visualizing the Effect of Process Pause on Virus Entrapment During Constant Flux Virus Filtration.

Area of Science:

• Biopharmaceutical Manufacturing
• Filtration Technology
• Virology

Background:

• Virus filtration is a critical step in biopharmaceutical manufacturing to remove adventitious viruses.
• Achieving high virus reduction (e.g., 10,000-fold) is challenging due to similar sizes of monoclonal antibodies and virus particles.
• Minute virus of mice (MVM) is a standard model virus for these studies.

Purpose of the Study:

• To visualize and understand virus entrapment within filtration membranes.
• To investigate the impact of process pauses and protein aggregates on virus filtration performance.
• To enhance the understanding of virus filtration membrane behavior.

Main Methods:

• Labeling Minute virus of mice (MVM) with a fluorescent dye.
• Utilizing laser scanning confocal microscopy for virus entrapment visualization.
• Testing three hollow fiber membranes (regenerated cellulose, polyvinylidene fluoride) with MVM in buffer and bovine serum albumin.

Main Results:

• A 30-minute process pause caused virus entrapment zones to broaden and move deeper into the membrane.
• The presence of bovine serum albumin aggregates led to a broader virus entrapment zone.
• Both process pauses and aggregates negatively impacted virus clearance by altering entrapment.

Conclusions:

• Visualization techniques provide insights into virus filtration mechanisms.
• Process parameters like pauses and feed composition (aggregates) significantly influence virus removal efficiency.
• Understanding entrapment dynamics is key to optimizing virus filtration strategies for biopharmaceuticals.