Factor IX

Grancha, Salvador; Herring, Steven W.; Páez, Antonio; Ristol, Pere; Jorquera, Juan I.

doi:10.1002/9781118356807.ch6

Cited by 3 publications

(1 citation statement)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…According to current regulations, at least two orthogonal virus clearance steps must be implemented to ensure viral safety of the final product [6]. The steps to mitigate virus contamination of FIX and PCC products include donor screening for known blood-borne viruses, i.e., HIV 1-2, HBV, HCV, HAV, and parvo B19; virus inactivation, such as solvent/detergent, mixed chemical inactivation (tri-n-butyl phosphate) and detergent (nonionic, polysorbate, and polyethylene oxide) treatment; and incubating intermediate product in controlled temperature (usually 6 h at 25 • C) [3,7,8]. Dry heat treatment may also be used after lyophilization, e.g., 100 • C for 1 h or 80 • C for 72 h. In the past, steam treatment at 60 • C (190 mbar) for 10 h or 80 • C (375 mbar) for 1 h was reported [3,7,8].…”

Section: Introductionmentioning

confidence: 99%

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

Manukyan

Mantas

Razumikhin³

et al. 2020

Biomedicines

View full text Add to dashboard Cite

Coagulation Factor IX-rich protrhombin complex concentrate (FIX-PCC) is a therapeutic biologic product that consists of a mixture of several human plasma-derived proteins, useful for treating hemophilia B. Due to its complex composition, FIX-PCC is very challenging to bioprocess through virus removing nanofilters in order to ensure its biosafety. This article describes a two-step filtration process of FIX-PCC using a nanocellulose-based filter paper with tailored porosity. The filters were characterized with scanning electron microscopy (SEM), cryoporometry with differential scanning calorimetry, and nitrogen gas sorption. Furthermore, in order to probe the filter’s cut-off size rejection threshold, removal of small- and large-size model viruses, i.e., ΦX174 (28 nm) and PR772 (70 nm), was evaluated. The feed, pre-filtrate, and permeate solutions were characterized with mass-spectrometric proteomic analysis, dynamic light scattering (DLS), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and analytical size-exclusion high-performance liquid chromatography (SEHPLC). By sequential filtration through 11 μm pre-filter and 33 μm virus removal filter paper, it was possible to achieve high product throughput and high virus removal capacity. The presented approach could potentially be applied for bioprocessing other protein-based drugs.

show abstract

Section: Introductionmentioning

confidence: 99%

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

Manukyan

Mantas

Razumikhin³

et al. 2020

Biomedicines

View full text Add to dashboard Cite

show abstract

Purification of coagulation factor VIII by immobilized metal affinity chromatography

Rodrigues

Verinaud

Oliveira

et al. 2014

Biotech and App Biochem

View full text Add to dashboard Cite

Factor VIII (FVIII) is a glycoprotein that plays an essential role in blood coagulation cascade. Purification of plasma-derived coagulation FVIII by direct application of plasma to a chromatographic column is a method of choice. Anion exchange column is a very powerful method because FVIII is strongly adsorbed, resulting in good activity recovery and high purification factor. However, vitamin-K-dependent coagulation factors coelute with FVIII. In the present study, we report the separation of vitamin-K-dependent coagulation proteins from FVIII using immobilized metal affinity chromatography (IMAC) with Cu(2+) as the metal ligand. Plasma was directly loaded to a Q Sepharose Big Beads column, and FVIII was recovered with 65% activity and a purification factor of approximately 50 times. Then, the Q Sepharose eluate was applied to the IMAC-Cu(2+) column, and FVIII was eluted with 200 mM imidazole, with up to 85% recovery of activity. The mass recovery in this fraction was less than 10% of the applied mass of protein. Vitamin-K-dependent proteins elute with imidazole concentrations of lower than 60 mM. Because of the difference in affinity, FVIII could be completely separated from the vitamin-K-dependent proteins in the IMAC column.

show abstract