Production of rhEPO with a serum-free medium in the packed bed bioreactor.
Recombinant CHO (C2) cells producing human erythropoietin (rhEPO) were cultured with DMEM:F12 media containing 5% FBS for 8-10 days in a packed bed bioreactor, then rhEPO was produced with a serum-free medium (SFM-p) which was prepared in our laboratory. The SFM-p medium can support the growth of C2 cells and the production of rhEPO, and furthermore, it easily separates rhEPO from the culture supernatant. The cell culture in a packed bed bioreactor system using SFM-p was maintained in a stable condition for 20-25 days. The expression level of rhEPO was 12-28.4 mg/L. The bioreactor productivity was 71.0 mg/L.d and increased by 12-14 fold over that of the roller bottle. The glucose consumption rate was 21 g/L.d. At the end of 30 days of perfusion circulation, a final cell density of over 3.0 x 10(7)/ml of culture volume was achieved. Since the cells were entrapped in the polyester disk, the culture supernatant contained only a few detachment cells. Variations in lactate and ammonia production in the reactor were observed, and results showed that the productions of lactate and ammonia by the bioreactor were 3.5 g/L and 5 mmol/L, respectively, and did not affect the expression of interest protein. This experiment demonstrates that SFM-p is suitable for the growth and rhEPO production of recombinant C2 in the packed bed bioreactor.
Deng
J
J
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing, China.
Yang
Q
Q
Cheng
X
X
Li
L
L
Zhou
J
J
D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D018929:Cell Culture Techniques; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D004921:Erythropoietin / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D016895:Culture Media, Serum-Free; D011993:Recombinant Fusion Proteins; D004921:Erythropoietin