Nanocellulose-Based Nanoporous Filter Paper for Virus Removal Filtration of Human Intravenous Immunoglobulin

Abstract

Human intravenous immunoglobulin (IVIG) is a highly valuable plasma-derived biotherapeutic with several important clinical indications in primary and acquired immunodeficiencies as well as autoimmune diseases, especially neuropathies. Ensuring the viral safety of plasma-derived products, such as human IVIG, is mandatory. Viral filtration is commonly used to affect viral removal in the manufacture of plasma products. Viral filtration of large volumes of a IVIG feed solution can take significant time, the required filter area can be large, and the resultant total cost of filtration is considerable. Therefore, there is a need for a high-capacity filter, which can process large volumes of plasma-derived biotherapeutic products within a short time at reduced cost. Here, we describe for the first time the performance of a nanocellulose-based virus removal filter paper in the processing of human IVIG, which has the potential to address the above-stated issues. The filter exhibited 5–6 log virus clearance of ΦX174 (28 nm; pI 6.6) or MS2 (27 nm; pI 3.9) phages during the filtration of spiked IVIG solutions (11 mg/mL, pH 4.9). To simulate real-life production conditions, filtration at 288 L/m2, corresponding to 3 kg of protein/m2, at 3 bar was undertaken. No substantial filter fouling was evident, with the flux remaining stable throughout filtration at 20–30 L/m2·h. The predicted volumetric capacity V max was ≥1700 L/m2, which corresponds to the processing of ≥19 kg/m2 of immunoglobulins. A number of characterization tests encompassing size-exclusion high-pressure liquid chromatography, dynamic light scattering, and polyacrylamide gel electrophoresis confirmed immunoglobulin integrity before and after filtration. This study has shown that a mille-feuille filter paper manufacturing process offers the possibility of producing cost-efficient viral removal filters with the required performance capabilities suitable for the processing of plasma-derived immunoglobulins and recombinant monoclonal antibodies.

Would 20 nm Filtered Fetal Bovine Serum-Supplemented Media Support Growth of CHO and HEK-293 Cells?

“…For brevity, the detailed procedures for filter-making and characterization can be found in previous publications from our group in open access sources. 24 , 25 , 27 …”

“…For brevity, the detailed procedures for listed proteomic characterizations can be found in previous publications from our group in open access sources. 24 , 25 , 27 Additionally, for the determination of levels of specific protein biomarkers, i.e. LC3B and β-actin, Western blotting was performed with immunoblotting antiobodies.…”

Emerging development of nanocellulose as an antimicrobial material: an overview

“…Size-exclusion-based virus filtration is an established, reliable and robust method to physically remove all types of infective microorganisms, including viruses. 64 According to Kosiol et al (2019), 65 size-exclusion-based virus filtration has a complex internal pore network, which is capable of transmitting molecules that are smaller than 15 nm on a quantitative basis. Therefore, at least 99.99% of the larger microorganisms, including viruses (or the majority of relevant virus species), should be retained, mainly by size exclusion.…”

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

“…The filter paper was previously validated in numerous studies to remove several large and small-size model viruses, including retroviruses (xMuLV, 100 nm) [35], parvoviruses (MVM, 20 nm) [32,36], and model phages (ΦX174, 28 nm) [37][38][39]. Recently, it was shown that this nanocellulose-based virus removal filter paper is useful for bioprocessing human plasma-derived IgG [40].…”