SUPERMACROPORUS CRYOGEL SYSTEM FOR CULTURING HYBRIDOMA CELLS
B. Premkumar*, N. S. Jayaprakash and M. A. Vijayalakshmi
ABSTRACT
In this experimental study, a mini-bioreactor setup was developed with cryogel matrix (35.32ml bed volume) for hybridoma cell H9E10 secreting antibodies against β-lactoglobulin are immobilized and evaluated for mAb production. The hybridoma cells were inoculated inside the gelatin-cryogel column by passing the cell suspension (25ml; 2.2 x 107). The initial flow through of about 6ml was collected which contained about 6.6 x 105 cells. The column retained about 19ml of the cell suspension which contained a total cell number of about 2.134 x 107 cells. The cells were left to grow and to establish themselves in the cryogel matrix for 12 h. After 12 h of incubation, we observed that the flow through (25ml) contained 9.2 x 105cells, which indicated that most of the cells got immobilized. A very few cells were detected in the flow through of the column after 48 h and as the cells were bound on the cryogel support matrix, the media was continuously circulated through the mini-bioreactor at a flow rate of 0.6 ml/min. The cryogel reactor was continuously run for a total period of 26 days until the gel became loose and cells started releasing out of the gel matrix.
Keywords: Supermacroporous Cryogel Matrix, Monoclonal Antibody.
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