Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor
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- 作者：Suthasinee Nilsang ; Kutty Selva Nandakumar ; Igor Yu. Galaev ; Sudip Kumar Rakshit ; Rikard Holmdahl ; Bo Mattiasson ; Ashok Kumar
- 刊名：Biotechnology Progress
- 出版年：2007
- 出版时间：August 2007
- 年：2007
- 卷：23
- 期：4
- 页码：932 - 939
- 全文大小：378K
- 年卷期：v.23,no.4(August 2007)
- ISSN：1520-6033
文摘
A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secretingantibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II,are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cellsgot attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside thecryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulationmedium. Glucose consumption and lactic acid production were also monitored, and during theexponential phase (~20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose,produced 2.48 mM day-1 lactic acid, and produced 6.5 
g mL-1 day-1 mAb during theexponential phase. The mAb concentration reached 130 g mL-1 after continuous run of thecryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, whichwas three times greater than the mAb yield obtained from T-flask batch cultivation. Even afterthe exchange of medium reservoir, cells in the cryogel column were still active and had relativelystable mAb production for an extended period of time. The bioreactor was operated continuouslyfor 55 days without any contamination. The results from ELISA as well as arthritis experimentsdemonstrate that the antibodies secreted by cells grown on the cryogel column did not differfrom antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus,supermacroporous cryogels can be useful as a supporting material for productive hybridomacell culture. Cells were found to be viable inside the porous matrix of the cryogel during thestudy period and secreted antibodies continuously. The antibodies thus obtained from the cryogelreactor were found to be functionally active in vivo, as demonstrated by their capacity to inducearthritis in mice.
g mL-1 day-1 mAb during theexponential phase. The mAb concentration reached 130 g mL-1 after continuous run of thecryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, whichwas three times greater than the mAb yield obtained from T-flask batch cultivation. Even afterthe exchange of medium reservoir, cells in the cryogel column were still active and had relativelystable mAb production for an extended period of time. The bioreactor was operated continuouslyfor 55 days without any contamination. The results from ELISA as well as arthritis experimentsdemonstrate that the antibodies secreted by cells grown on the cryogel column did not differfrom antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus,supermacroporous cryogels can be useful as a supporting material for productive hybridomacell culture. Cells were found to be viable inside the porous matrix of the cryogel during thestudy period and secreted antibodies continuously. The antibodies thus obtained from the cryogelreactor were found to be functionally active in vivo, as demonstrated by their capacity to inducearthritis in mice.