Recombinant Antibody Production by Perfusion Cultures of rCHO Cells in a Depth Filter Perfusion System

Abstract

Recombinant Chinese hamster ovary cells, producing recombinant antibody against the human platelet, were cultivated in a depth filter perfusion system (DFPS). When perfusion cultures with working volume of 1 L were operated at perfusion rates of 5/d and 6/d, volumetric antibody productivities reached values 28 and 34 times higher than that of batch suspension culture in Erlenmeyer flasks and 43 and 53 times higher than that of batch culture in a controlled stirred tank reactor, respectively. Perfusion cultures in the DFPS showed stable antibody production over the whole culture period of up to 20 days. In the DFPS, inoculated cells in suspension were entrapped in a few hours within the depth filter matrix by medium circulation and retained there until the void space of the filter matrix was saturated by the cultured cells. After cells in the depth filter matrix reached saturation, overgrown viable cells at a perfusion rate of 5/d or 6/d were continuously collected into waste medium at a density of 2-4 x 10(5) cells/mL, which resulted in stable operation at high perfusion rates, maintaining values of process parameters such as glucose/lactate concentration, pH, and dissolved oxygen concentration. Because the DFPS overcomes most drawbacks observed with conventional perfusion systems, it is preferable to be used as a key culture system to produce monoclonal antibody stably for a long culture period.

Multi-stage high cell continuous fermentation for high productivity and titer

“…The productivity ratio (PR) of monoclonal antibody (Mab) was as high as 10 since the production was not inhibited by Mab itself but by lactic acid and ammonia. This particular Mab MSC-HCDC system does not need a stage-wise cell recycling since the cells are immobilized in the depth filter perfusion system (DFPS) of each stage [50][51][52]. The results obtained in this study suggest a possibility that the MSC-HCDC system can significantly reduce the production cost and plant size for the production of almost all fermentation products.…”

Dynamic Modeling of Apoptosis and its Interaction with Cell Growth in Mammalian Cell Culture

“…Operation of the cell culture process in either fed-batch or perfusion modes is another way of enhancing cell density and thus productivity (Detremblay et al, 1992;Lee et al, 2005;Nguang et al, 2001;Trampler et al, 1994). With these modes of operation, it is possible to reach higher cell density and prolonged sustainability of cell viability.…”

Two-stage depth filter perfusion culture for recombinant antibody production by recombinant Chinese hamster ovary cell

“…A single stage DFPS was described in detail in a previous study by the present authors [4]. The depth filter used in the present study was a Profile II TM depth filter from Pall Corp. (East Hills, NY, USA) and was used as a filter matrix for cell entrapment.…”

“…In our previous studies, the efficiency of DFPSs was successfully investigated for high-volumetric productivity of recombinant antibody [4][5][6][7]. With greatly increased density of animal cells, the volumetric productivity of recombinant antibody reached 72.3 mg/L/d, which was 53 times that of the conventional batch process.…”

“…Contemporary [G1] perfusion systems have drawbacks such as uneasy sustenance for long-term production. However, the DFPS [G2] showed operational stability without filter clogging, and continuous cell bleeding occurred even after the filter had been saturated with the cells [4].…”