Producing Ad-IFN gamma by suspension culture of HEK293 cells in a disposable bioreactor / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1786-1790, 2014.
Artículo
en Chino
| WPRIM
| ID: wpr-345544
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.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Cultivo de Virus
/
Terapia Genética
/
Recuento de Células
/
Adenoviridae
/
Reactores Biológicos
/
Células HEK293
/
Técnicas de Cultivo Celular por Lotes
/
Vectores Genéticos
/
Métodos
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2014
Tipo del documento:
Artículo
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