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

Levon Manukyan¹, Athanasios Mantas¹, Mikhail Razumikhin²

¹Nanotechnology and Functional Materials, Department of Materials Science and Engineering, Uppsala University, Box 534, 751 21 Uppsala, Sweden.

|April 1, 2020

Summary

A novel two-step filtration method using nanocellulose filters efficiently removes viruses from Coagulation Factor IX-rich prothrombin complex concentrate (FIX-PCC). This bioprocessing advance enhances biosafety for hemophilia B treatment and potentially other protein-based therapies.

Keywords:

Cladophora cellulose Mille-feuille filter hemophilia B protein aggregates virus removal filtration

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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.