engleski

Adaptation of CHO cells in serum-free conditions for erythropoietin production: Application of EVOP for process optimization

Mammalian cell cultures are the preferred expression systems for the production of biopharmaceuticals requiring post-translational processing. Usually, cell cultures are cultivated in medium supplemented with serum, which supports cell proliferation, viability and productivity. However, due to scientific and regulatory concerns, serum-free conditions are required in recombinant protein production. Cell lines that are intended for commercial recombinant protein production have to be adapted to serum- or protein- free conditions early in their development. This is a labor- and time- consuming process because of the specific cell requirements related to their adaptation in new microenvironment. In the present study, a Chinese hamster ovary (CHO) cell line producing glycosylated recombinant human erythropoietin (rhEPO) was adapted for growth and rhEPO production in serum-free and protein-free conditions. The physiology, growth parameters and morphology of the CHO cells and rhEPO biosynthesis and structure were closely monitored during the adaptation process to avoid unwanted selection of cell subpopulations. The results showed that the CHO cells were successfully adapted to suspension growth and rhEPO production in the protein free conditions and that the structure of rhEPO remained nearly unchanged. In addition, during rhEPO production in the protein-free suspension conditions, the agitation rate showed to be significant for optimal process performance in contrast to the initial cell concentration, evaluated through evolutionary operation (EVOP) method.

CHO cells ; protein - free medium ; erythropoietin ; EVOP ; suspension culture

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