Producing Ad-IFN gamma by suspension culture of HEK293 cells in a disposable bioreactor.
- Author:
Quande WU
;
Wenlin HUANG
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
growth & development;
Batch Cell Culture Techniques;
Bioreactors;
Cell Count;
Genetic Therapy;
Genetic Vectors;
HEK293 Cells;
Humans;
Virus Cultivation;
methods
- From:
Chinese Journal of Biotechnology
2014;30(11):1786-1790
- CountryChina
- Language:Chinese
-
Abstract:
Adenovirus vectors are promising delivery systems for gene therapy. We established a new process for clinic trial of recombinant adenovirus vectors using a novel disposable bioreactor. The suspension HEK293 cells were inoculated into a 5 L disposable bioreactor with parameters control of pH, DO, agitation and temperature. After 6 days of a fed-batch culture, the final cell density reached 2.0 x 10(6) cells/mL. The culture was infected with Ad-IFNγ at an MOI of 30. The harvest was performed at approximately 48 h post-infection and crude viral lysate was obtained after 3 freeze/thaw cycles and centrifugation. The maximum titers of crude viral lysate was 1.49 x 10(13) Infectious units (IFU) and the bulk product specific was 3,800 IFU/cell. Purified Ad-IFNγ by anion-exchange chromatography and the final recovery of infectious unit reached 35.9%. The result demonstrates that an efficient and stable process of producing Ad-IFNγ using a disposable fed-batch bioreactor is established.
