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Continuous production of monoclonal antibody in a packed‐bed bioreactor
Author(s) -
Golmakany Naghmeh,
Rasaee Mohammad Javad,
Furouzandeh Mehdi,
Shojaosadati Seyed Abbas,
Kashanian Soheila,
Omidfar Kobra
Publication year - 2005
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20040121
Subject(s) - monoclonal antibody , bioreactor , dilution , chromatography , cell culture , yield (engineering) , liter , matrix (chemical analysis) , chemistry , lactate dehydrogenase , fed batch culture , microbiology and biotechnology , biology , antibody , biochemistry , materials science , enzyme , fermentation , immunology , physics , genetics , organic chemistry , metallurgy , endocrinology , thermodynamics
In the present study the growth and MAb (monoclonal antibody) production of a mouse×mouse hybridoma cell producing anti‐digoxin MAb was evaluated. The hybridoma cells entrapped within the support matrix Fibra‐Cel® were cultured in batch and continuous mode following special protocols. Cell‐culture studies were performed in a 1‐litre spinner basket containing 3 g·litre −1 support matrix. Batch culture was operated with the cell density of 42×10 6 cells. During the 7 days of culture, the medium was sampled daily in order to assess glucose and MAb concentrations and the lactate dehydrogenase released into the culture medium. After a culture period of 72 h, the cell density and MAb concentration were found to be 10.4×10 7 cells/3 g of NWPF (non‐woven polyester fibre) discs and 250 μg/ml respectively. This yield gradually decreased to 0.55×10 6 cells/3 g of packaging material and 60 μg/ml respectively at the end of the batch culture. In the continuous‐culture studies, the batch culture was initially operated for 64.5 h and then continuous flow was started at the dilution rates of 0.15, 0.2, 0.25 and 0.3 day −1 and finally stabilized at 0.25 day −1 within 288 h (12 days). The MAb concentration at steady state was found to be 116–120 μg/day per ml, and the yield of operation was 62.5 mg/day per ml, which was 3.5 times higher than that of batch culture. In conclusion, a packed‐bed bioreactor with the support matrix Fibra‐Cel®, operated in continuous‐feeding mode, is more efficient for large‐scale MAb production than a batch culture. On the other hand, by using a continuous‐culture system, a better supply of nutrients and removal of inhibitory metabolites and proteolytic enzymes was obtained.