Two-Step Size-Exclusion Nanofiltration of Prothrombin Complex Concentrate Using Nanocellulose-Based Filter Paper

Area of Science:

• Biotechnology
• Bioprocessing
• Filtration Technology

Background:

• Coagulation Factor IX-rich prothrombin complex concentrate (FIX-PCC) is crucial for hemophilia B treatment.
• The complex protein composition of FIX-PCC poses significant challenges for virus removal during bioprocessing, impacting biosafety.
• Ensuring the viral safety of therapeutic biologics like FIX-PCC is paramount.

Purpose of the Study:

• To develop and evaluate a novel two-step filtration process for effective virus removal from FIX-PCC.
• To characterize nanocellulose-based filter papers with tailored porosity for bioprocessing applications.
• To assess the capacity of the filtration system for high product throughput and virus removal.

Main Methods:

• Utilized nanocellulose-based filter papers with specifically engineered porosity for sequential filtration.
• Characterized filter properties using scanning electron microscopy (SEM), cryoporometry, and nitrogen gas sorption.
• Evaluated virus removal efficiency using model viruses (ΦX174, PR772) and characterized filtrates via proteomic analysis, DLS, SDS-PAGE, and SEHPLC.

Main Results:

• A two-step filtration employing an 11 μm pre-filter and a 33 μm virus removal filter paper demonstrated high product throughput.
• The nanocellulose filters effectively removed both small and large model viruses, confirming high virus removal capacity.
• Filtration process parameters and filter characteristics were thoroughly analyzed using multiple analytical techniques.

Conclusions:

• The developed two-step filtration process using tailored nanocellulose filters is effective for virus removal from FIX-PCC.
• This approach significantly enhances the biosafety of FIX-PCC bioprocessing.
• The filtration strategy holds potential for application in the bioprocessing of other complex protein-based therapeutic products.