September 17-21, 2017
The main goal of this study was to develop an innovative CHO-based process for the production of glucocerebrosidase (GBA), an enzyme used for the replacement therapy of Type 1 Gaucher disease. The focus of the present study was on the development of a perfusion process, combining strategies that are commonly used for process optimization: temperature reduction, and supplementation of the culture medium with productivity enhancers, such as short chain fatty acids. The effects of mild hypothermic conditions combined with valeric acid supplementation were first studied in batch shake flasks for two clones (CHO-GBA-36K and CHO-GBA-65P), developed previously using as host the cell lines CHO.K1 (ATCC CCL-61) and CHO.PRO5 (a glycosylation mutant developed by Stanley et al. Cell 6:121, 1975), respectively. A DOE approach was used (Table 1) to select the most promising cultivation conditions to be further applied to a perfusion process. The best performance regarding both cell growth and GBA production was obtained for the CHO-GBA-65P clone under condition , at 31ºC with no valeric acid (Table 1). Under this condition, CHO-GBA-65P achieved a maximum qP of 58.4 mU/106 cells/d, which is 4.2 fold higher than qP at the control condition  and 2.7 fold higher than the maximum qP obtained for the CHO-GBA-36K clone, which was achieved at 31ºC with 2 mM valeric acid supplementation (condition ).
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Filipa M. Gonçalves, Leda R. Castilho, and Juliana Coronel, "Use of a biphasic perfusion process based on mild hypothermia for recombinant glucocerebrosidase (GBA) production" in "Integrated Continuous Biomanufacturing III", Suzanne Farid, University College London, United Kingdom Chetan Goudar, Amgen, USA Paula Alves, IBET, Portugal Veena Warikoo, Axcella Health, Inc., USA Eds, ECI Symposium Series, (2017). https://dc.engconfintl.org/biomanufact_iii/87