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Effect of transmembrane pressure on Factor VIII yield in ATF perfusion culture for the production of recombinant human Factor VIII co-expressed with von Willebrand factor
- Source :
- Cytotechnology. 68:1687-1696
- Publication Year :
- 2015
- Publisher :
- Springer Science and Business Media LLC, 2015.
-
Abstract
- In this study, we evaluated three cell retention devices, an alternating tangential flow (ATF) system, a spin-filter, and a Centritech Lab III centrifuge, for the production of recombinant human Factor VIII co-expressed with von Willebrand factor. From the results, it was found that the FVIII activity in bioreactor was significantly higher in the ATF perfusion culture than two other perfusion cultures. Moreover, the FVIII activity yield was unexpectedly low in the ATF perfusion culture. We have, therefore, studied the reasons for this low FVIII activity yield. It was revealed that the inactivation and the surface adsorption of FVIII onto the harvest bag were not the main reasons for the low yield in the ATF perfusion culture. The FVIII activity yield was not increased by the use of a hollow fiber filter with 0.5 μm pore size instead of 0.2 μm pore size. Additionally, the retention of FVIII molecules by the hollow fiber filter was a dominant factor in the low FVIII activity yield in the ATF perfusion culture. We demonstrated that FVIII yield was significantly improved by controlling transmembrane pressure (TMP) across the hollow fiber filter membrane. Taken together, these results suggest that TMP control could be an efficient method for the enhancement of FVIII yield in an ATF perfusion culture.
- Subjects :
- 0301 basic medicine
congenital, hereditary, and neonatal diseases and abnormalities
animal diseases
Recombinant human factor VIII
Clinical Biochemistry
Cell
Biomedical Engineering
Bioengineering
Transmembrane pressure
03 medical and health sciences
Perfusion Culture
Von Willebrand factor
hemic and lymphatic diseases
medicine
Bioreactor
biology
Chemistry
Cell Biology
030104 developmental biology
medicine.anatomical_structure
Yield (chemistry)
Biophysics
biology.protein
Original Article
Perfusion
Biotechnology
Subjects
Details
- ISSN :
- 15730778 and 09209069
- Volume :
- 68
- Database :
- OpenAIRE
- Journal :
- Cytotechnology
- Accession number :
- edsair.doi.dedup.....f0d5ed63e98db4c1aa4f868d9bbed999