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Kaneko Y.,Division 5 Technology | Kaneko Y.,University of Tsukuba | Sato R.,Chugai Pharmaceutical Manufacturing Co. | Aoyagi H.,University of Tsukuba
Journal of Bioscience and Bioengineering | Year: 2010

Pharmaceutical manufacturing plants can be operated continuously for several months. It is therefore important to use cells with long-term stability for the production of active ingredients. We investigated the reliability and long-term stability of an antibody-producing cell line. A recombinant Chinese hamster ovary (CHO) cell line was cultivated in spinner flasks and reactors, including a practical production-scale reactor (1600 L), for 109 days to produce monoclonal antibodies against the HM1.24 antigen. During cultivation, the cells remained stable and there was an increase in the rate of cell proliferation, yielding viable cells at high density. A decrease in cell-specific productivity was associated with this increase in the rate of cell proliferation. The cells were genetically stable and other measures of cellular function remained consistent throughout the cultivation period. © 2009 The Society for Biotechnology, Japan.


Kaneko Y.,Division 5 Technology | Kaneko Y.,University of Tsukuba | Sato R.,Chugai Pharmaceutical Manufacturing Co. | Aoyagi H.,University of Tsukuba
Journal of Bioscience and Bioengineering | Year: 2010

Although overproduction of recombinant proteins by mammalian cells is well established, little attention has been paid to analysis of the quality of the products. We focused on the quality of antibodies produced during the death phase of a recombinant Chinese hamster ovary (CHO) cell line. The quality of the monoclonal antibody against HM1.24 antigen and the post-translational characteristics of the subunits during CHO cell culture in a 160-L bioreactor were investigated. The culture supernatant of a stable cell line was collected and purified by affinity chromatography and then analyzed. There were no significant changes in gel-permeation chromatography variables, carbohydrate structure, or antibody-dependent cellular cytotoxicity activity during the death phase of cell culture. However, ion-exchange chromatography analysis revealed that antibody heterogeneity changed, as indicated by a decrease in cell viability. The results presented here provide useful information that will help in determining the time to end each batch culture. © 2009 The Society for Biotechnology, Japan.


Kishishita S.,Chugai Pharmaceutical Co. | Kishishita S.,University of Tsukuba | Katayama S.,Chugai Pharmaceutical Co. | Kodaira K.,Division 5 Technology | And 6 more authors.
Journal of Bioscience and Bioengineering | Year: 2015

Chinese hamster ovary (CHO) cells are the most commonly used mammalian host for large-scale commercial production of therapeutic monoclonal antibodies (mAbs). Chemically defined media are currently used for CHO cell-based mAb production. An adequate supply of nutrients, especially specific amino acids, is required for cell growth and mAb production, and chemically defined fed-batch processes that support rapid cell growth, high cell density, and high levels of mAb production is still challenging. Many studies have highlighted the benefits of various media designs, supplements, and feed addition strategies in cell cultures. In the present study, we used a strategy involving optimization of a chemically defined feed medium to improve mAb production. Amino acids that were consumed in substantial amounts during a control culture were added to the feed medium as supplements. Supplementation was controlled to minimize accumulation of waste products such as lactate and ammonia. In addition, we evaluated supplementation with tyrosine, which has poor solubility, in the form of a dipeptide or tripeptide to improve its solubility. Supplementation with serine, cysteine, and tyrosine enhanced mAb production, cell viability, and metabolic profiles. A cysteine-tyrosine-serine tripeptide showed high solubility and produced beneficial effects similar to those observed with the free amino acids and with a dipeptide in improving mAb titers and metabolic profiles. © 2014 The Society for Biotechnology, Japan.

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